U.S. patent application number 12/403824 was filed with the patent office on 2009-10-08 for method and apparatus for irradiating a surface with pulsed light.
Invention is credited to Luis De Taboada, Jackson Streeter.
Application Number | 20090254154 12/403824 |
Document ID | / |
Family ID | 41133964 |
Filed Date | 2009-10-08 |
United States Patent
Application |
20090254154 |
Kind Code |
A1 |
De Taboada; Luis ; et
al. |
October 8, 2009 |
METHOD AND APPARATUS FOR IRRADIATING A SURFACE WITH PULSED
LIGHT
Abstract
A method and apparatus irradiates a surface with at least one
pulsed light beam emitted from an emission surface of an optical
element. The at least one pulsed light beam comprises a plurality
of pulses having a temporal pulsewidth in a range between about 0.1
millisecond and about 150 seconds. The at least one pulsed light
beam has a beam cross-sectional area at the emission surface
greater than about 2 cm.sup.2 and a time-averaged irradiance in a
range between about 1 mW/cm.sup.2 and about 100 W/cm.sup.2.
Inventors: |
De Taboada; Luis; (Carlsbad,
CA) ; Streeter; Jackson; (Reno, NV) |
Correspondence
Address: |
KNOBBE MARTENS OLSON & BEAR LLP
2040 MAIN STREET, FOURTEENTH FLOOR
IRVINE
CA
92614
US
|
Family ID: |
41133964 |
Appl. No.: |
12/403824 |
Filed: |
March 13, 2009 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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12389294 |
Feb 19, 2009 |
|
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12403824 |
|
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61037668 |
Mar 18, 2008 |
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Current U.S.
Class: |
607/88 ;
607/110 |
Current CPC
Class: |
A61N 5/0613 20130101;
A61N 2005/007 20130101; A61N 2005/0644 20130101; A61N 2005/0647
20130101; A61N 2005/0659 20130101 |
Class at
Publication: |
607/88 ;
607/110 |
International
Class: |
A61N 5/06 20060101
A61N005/06; A61F 7/00 20060101 A61F007/00 |
Claims
1. An apparatus for irradiating a portion of a patient's scalp or
skull with light, the apparatus comprising: a source of light; and
an output optical element in optical communication with the source,
wherein the output optical element comprises an emission surface
configured to emit a pulsed light beam comprising a plurality of
pulses having a temporal pulsewidth in a range between 0.1
millisecond and 150 seconds, the pulsed light beam having a
cross-sectional area greater than about 2 cm.sup.2 at the emission
surface of the output optical element, having a time-averaged
irradiance in a range of about 10 mW/cm.sup.2 to about 10
W/cm.sup.2 across the cross-sectional area.
2. The apparatus of claim 1, wherein the pulsed light beam
stimulates, excites, induces, or otherwise supports one or more
intercellular or intracellular biological processes involved in the
survival, regeneration, or restoration of performance or viability
of brain cells.
3. The apparatus of claim 1, wherein pulsed light beam has a duty
cycle, wherein the temporal pulsewidth and the duty cycle are
sufficient for the pulsed light beam to penetrate the skull to
modulate membrane potentials, thereby enhancing cell survival, cell
function, or both of the irradiated brain cells.
4. The apparatus of claim 1, wherein the pulsed light beam at the
emission surface has a beam diameter in a range between 10
millimeters and 40 millimeters, an average irradiance per pulse in
a range between 10 mW/cm.sup.2 and 10 W/cm.sup.2, one or more
wavelengths in a range between 600 nanometers and 980 nanometers,
and a duty cycle in a range between 10% and 30%.
5. A method of irradiating a surface with light, the method
comprising: irradiating the surface with at least one pulsed light
beam emitted from an emission surface of an optical element, the at
least one pulsed light beam comprising a plurality of pulses having
a temporal pulsewidth in a range between about 0.1 millisecond and
about 150 seconds, the at least one pulsed light beam having a beam
cross-sectional area at the emission surface greater than about 2
cm.sup.2 and a time-averaged irradiance in a range between about 1
mW/cm.sup.2 and about 100 W/cm.sup.2.
6. The method of claim 5, wherein the beam cross-sectional area at
the emission surface is in a range between about 1 cm.sup.2 and
about 20 cm.sup.2.
7. The method of claim 5, wherein the average irradiance per pulse
at the emission surface is in a range between about 10 mW/cm.sup.2
and about 10 W/cm.sup.2.
8. The method of claim 5, wherein the average irradiance per pulse
at the emission surface is in a range between about 100 mW/cm.sup.2
and about 1 W/cm.sup.2.
9. The method of claim 5, wherein the temporal pulsewidth is in a
range between about 0.1 millisecond and about 100 milliseconds.
10. The method of claim 5, wherein the at least one pulsed light
beam has one or more wavelengths are in a range between about 600
nanometers and about 980 nanometers.
11. The method of claim 5, wherein the at least one pulsed light
beam has a duty cycle in a range between about 1% and about
80%.
12. The method of claim 5, wherein the at least one pulsed light
beam has a beam divergence angle at the emission surface greater
than zero and less than 35 degrees.
13. The method of claim 5, wherein the irradiated surface comprises
at least a portion of the scalp or skull of a patient whose brain
has experienced at least one physical trauma, at least a portion of
the at least one pulsed light beam transmitted through the skull to
irradiate at least a portion of the patient's brain.
14. The method of claim 13, wherein the irradiated surface
comprises an irradiated portion of the scalp, and the irradiated
portion of the scalp is not blanched while irradiating the portion
of the scalp.
15. The method of claim 13, wherein the irradiated surface
comprises an irradiated portion of the scalp, and the irradiated
portion of the scalp is blanched while irradiating the portion of
the scalp.
16. The method of claim 13, wherein the irradiated surface is
cooled while irradiating the irradiated surface.
17. The method of claim 13, wherein the irradiated surface is not
cooled while irradiating the irradiated surface.
18. The method of claim 13, wherein the irradiated surface
comprises an irradiated portion of the scalp, and the method
further comprises removing at least a portion of the hair from the
irradiated portion of the scalp prior to irradiating the irradiated
portion of the scalp.
19. The method of claim 13, wherein irradiating the surface
comprises sequentially irradiating a plurality of treatment sites
of the scalp with the pulsed light beam.
20. The method of claim 19, wherein the plurality of treatment
sites comprises at least 10 treatment sites.
21. The method of claim 19, wherein the plurality of treatment
sites comprises between 15 and 25 treatment sites.
22. The method of claim 13, wherein the average irradiance at the
scalp or skull is selected to provide an average irradiance greater
than 0.01 mW/cm.sup.2 at a depth of approximately 2 centimeters
below the dura.
23. The method of claim 13, wherein the light is transmitted
through a region of the patient's skull containing an excess amount
of hemorrhagic blood due to the at least one physical trauma.
24. The method of claim 5, wherein the irradiated surface comprises
a portion of a light-detection system configured to measure one or
more parameters of the pulsed light beam.
25. The method of claim 24, further comprising measuring the one or
more parameters of the pulsed light beam impinging the irradiated
surface.
26. A method of treating a patient's brain, the method comprising:
irradiating at least a portion of the scalp or skull of the patient
with at least one pulsed light beam comprising a plurality of
pulses transmitted through the patient's skull to irradiate at
least a portion of the brain, the at least one pulsed light beam
having a temporal profile comprising a time-averaged irradiance at
the scalp averaged over one second in a range between about 100
mW/cm.sup.2 and about 10 W/cm.sup.2 and a peak irradiance at the
scalp in a range between about 12.5 mW/cm.sup.2 and about 1000
W/cm.sup.2.
27. The method of claim 26, wherein the temporal profile does not
optimize the thermal relaxation of the irradiated tissue.
28. The method of claim 26, wherein the at least one pulsed light
beam has a beam cross-sectional area at the scalp greater than
about 2 cm.sup.2, and one or more wavelengths in a range between
about 780 nanometers and about 840 nanometers.
29. The method of claim 26, wherein the temporal profile further
comprises a temporal pulsewidth in a range between about 0.1
millisecond and about 300 milliseconds and a duty cycle in a range
between about 10% and about 30%.
30. The method of claim 26, wherein irradiating at least a portion
of the scalp or skull comprises sequentially irradiating a
plurality of treatment sites with a plurality of pulsed light
beams.
31. The method of claim 26, wherein irradiating the at least a
portion of the scalp or skull is performed for a time period of 60
seconds to 600 seconds.
32. A method of treating a patient who has experienced a traumatic
brain injury, the method comprising noninvasively irradiating at
least a portion of the patient's scalp or skull with pulsed light
penetrating the patient's skull to irradiate and stimulate brain
cells of the patient, the pulsed light having a temporal profile
comprising an average irradiance per pulse, a temporal pulse width,
and a pulse duty cycle, the temporal profile selected to modulate
membrane potentials in order to enhance, restore, or promote cell
survival, cell function, or both of the irradiated brain cells
following the traumatic brain injury.
33. The method of claim 32, wherein the average irradiance per
pulse is in a range between about 10 mW/cm.sup.2 and about 10
W/cm.sup.2.
34. The method of claim 32, wherein the pulsed light comprises a
light beam having a beam cross-sectional area in a range between 2
cm.sup.2 and 20 cm.sup.2.
35. The method of claim 32, wherein the temporal pulse width is in
a range between about 0.1 millisecond and about 300
milliseconds.
36. The method of claim 32, wherein the pulse duty cycle is in a
range between about 10% and about 30%.
37. A method of treating a patient experiencing a neurodegenerative
disease or depression, the method comprising noninvasively
irradiating at least a portion of the patient's scalp or skull with
pulsed light penetrating the patient's skull to irradiate and
stimulate brain cells of the patient, the pulsed light having a
temporal profile comprising an average irradiance per pulse, a
temporal pulse width, and a pulse duty cycle, the temporal profile
selected to modulate membrane potentials in order to enhance,
restore, or promote cell survival, cell function, or both of the
irradiated brain cells.
38. The method of claim 37, wherein the neurodegenerative disease
comprises Alzheimer's Disease or Parkinson's Disease.
Description
CLAIM OF PRIORITY
[0001] This application is a continuation-in-part from U.S. patent
application Ser. No. 12/389,294, filed Feb. 20, 2009, which is
incorporated in its entirety by reference herein. This application
also claims the benefit of priority to U.S. Provisional Application
No. 61/037,668, filed Mar. 18, 2008, which is incorporated in its
entirety by reference herein.
BACKGROUND OF THE INVENTION
[0002] 1. Field of the Invention
[0003] The present invention relates in general to phototherapy,
and more particularly, to novel apparatuses and methods for
phototherapy of brain tissue.
[0004] 2. Description of the Related Art
[0005] There are numerous neurologic conditions, such as
neurodegenerative diseases (e.g., Alzheimer's disease, Parkinson's
disease), Huntington's disease, demyelinating diseases (e.g.,
multiple sclerosis), cranial nerve palsies, traumatic brain injury,
stroke, and spinal cord injury which could possibly benefit from
application of phototherapy. Most of these conditions cause
significant morbidity and mortality and involve tremendous burden
to society, families and caregivers. Many neurologic conditions
have no currently available effective therapies or the therapies
that are available are not adequate to restore functional recovery,
sustain quality of life, or halt disease progression.
[0006] One example of a neurologic condition that remains a major
unmet medical need is stroke, also called cerebrovascular accident
(CVA). Stroke is caused by a sudden disruption of blood flow to a
discrete area of the brain that is brought on by the lodging of a
clot in an artery supplying blood to an area of the brain (called
an ischemic stroke), or by a cerebral hemorrhage due to a ruptured
aneurysm or a burst artery (called a hemorrhagic stroke). There are
over 750,000 stroke victims per year in the United States, and
approximately 85% of all strokes are ischemic and 15% are
hemorrhagic. The consequence of stroke is a loss of function in the
affected brain region and concomitant loss of bodily function in
areas of the body controlled by the affected brain region.
Depending upon the extent and location of the primary insult in the
brain, loss of function varies greatly from mild or severe, and may
be temporary or permanent. Lifestyle factors such as smoking, diet,
level of physical activity and high cholesterol increase the risk
of stroke, and thus stroke is a major cause of human suffering in
developed nations. Stroke is the third leading cause of death in
most developed nations, including the United States.
[0007] Stroke treatment is often restricted to providing basic life
support at the time of the stroke, followed by rehabilitation.
Currently, the only FDA-cleared treatment of ischemic stroke
involves thrombolytic therapy using tissue plasminogen activator
(tPA). However, tPA can only be used within three hours of stroke
onset and has several contraindications, therefore, only a small
percentage of stroke victims receive this drug.
[0008] Traumatic brain injury (TBI) occurs when a sudden physical
trauma (e.g. crush or compression injury in the central nervous
system, including a crush or compression injury of the brain,
spinal cord, nerves or retina, or any acute injury or insult
producing cell death) causes damage to the head. For example, a
sudden and/or violent blow to the head or an object piercing the
skull and entering brain tissue can result in TBI. The extent of
damage to the brain can be severe, however even mild and moderate
TBI has been associated with neurological sequelae that can be long
lasting. Development of neurodegenerative conditions has been
associated with TBI. TBI can result in a sudden disruption of blood
flow to a discrete area of the brain. The consequence of stroke or
TBI can be a loss of function in the affected brain region and
concomitant loss of bodily function in areas of the body controlled
by the affected brain region. Depending upon the extent and
location of the primary insult in the brain, loss of function
varies greatly from mild or severe, and may be temporary or
permanent.
[0009] A high level of interest and clinical need remains in
finding new and improved therapeutic interventions for treatment of
stroke and other neurologic conditions that continue to devastate
millions of lives each year and where few effective therapies
exist.
SUMMARY OF THE INVENTION
[0010] In certain embodiments, an apparatus for irradiating a
portion of a patient's scalp or skull with light is provided. The
apparatus comprises a source of light and an output optical element
in optical communication with the source. The output optical
element comprises an emission surface configured to emit a pulsed
light beam comprising a plurality of pulses having a temporal
pulsewidth in a range between 0.1 millisecond and 150 seconds. The
pulsed light beam has a cross-sectional area greater than about 2
cm.sup.2 at the emission surface of the output optical element, and
has a time-averaged irradiance in a range of about 10 mW/cm.sup.2
to about 10 W/cm.sup.2 across the cross-sectional area.
[0011] In certain embodiments, a method of irradiating a surface
with light is provided. The method comprises irradiating the
surface with at least one pulsed light beam emitted from an
emission surface of an optical element. The at least one pulsed
light beam comprises a plurality of pulses having a temporal
pulsewidth in a range between about 0.1 millisecond and about 150
seconds. The at least one pulsed light beam has a beam
cross-sectional area at the emission surface greater than about 2
cm.sup.2 and a time-averaged irradiance in a range between about 1
mW/cm.sup.2 and about 100 W/cm.sup.2.
[0012] In certain embodiments, a method of treating a patient's
brain is provided. The method comprises irradiating at least a
portion of the scalp or skull of the patient with at least one
pulsed light beam comprising a plurality of pulses transmitted
through the patient's skull to irradiate at least a portion of the
brain. The at least one pulsed light beam has a temporal profile
comprising a time-averaged irradiance at the scalp averaged over
one second in a range between about 100 mW/cm.sup.2 and about 10
W/cm.sup.2 and a peak irradiance at the scalp in a range between
about 12.5 mW/cm.sup.2 and about 1000 W/cm.sup.2.
[0013] In certain embodiments, a method of treating a patient who
has experienced a traumatic brain injury is provided. The method
comprises noninvasively irradiating at least a portion of the
patient's scalp or skull with pulsed light penetrating the
patient's skull to irradiate and stimulate brain cells of the
patient. The pulsed light has a temporal profile comprising an
average irradiance per pulse, a temporal pulse width, and a pulse
duty cycle. The temporal profile is selected to modulate membrane
potentials in order to enhance, restore, or promote cell survival,
cell function, or both of the irradiated brain cells following the
traumatic brain injury.
[0014] In certain embodiments, a method of treating a patient
experiencing a neurodegenerative disease or depression is provided.
The method comprises noninvasively irradiating at least a portion
of the patient's scalp or skull with pulsed light penetrating the
patient's skull to irradiate and stimulate brain cells of the
patient. The pulsed light has a temporal profile comprising an
average irradiance per pulse, a temporal pulse width, and a pulse
duty cycle. The temporal profile is selected to modulate membrane
potentials in order to enhance, restore, or promote cell survival,
cell function, or both of the irradiated brain cells.
[0015] For purposes of summarizing the present invention, certain
aspects, advantages, and novel features of the present invention
have been described herein above. It is to be understood, however,
that not necessarily all such advantages may be achieved in
accordance with any particular embodiment of the present invention.
Thus, the present invention may be embodied or carried out in a
manner that achieves or optimizes one advantage or group of
advantages as taught herein without necessarily achieving other
advantages as may be taught or suggested herein.
BRIEF DESCRIPTION OF THE DRAWINGS
[0016] FIG. 1 schematically illustrates an example beam delivery
apparatus in accordance with certain embodiments described
herein.
[0017] FIG. 2A schematically illustrates a cross-sectional view of
an example output optical assembly in accordance with certain
embodiments described herein.
[0018] FIG. 2B schematically illustrates another example output
optical assembly in accordance with certain embodiments described
herein.
[0019] FIGS. 3A and 3B schematically illustrate the diffusive
effect on the light by the output optical assembly.
[0020] FIGS. 4A and 4B schematically illustrate cross-sectional
views of two example beam delivery apparatuses in accordance with
certain embodiments described herein.
[0021] FIG. 5 schematically illustrates an example fiber alignment
mechanism in accordance with certain embodiments described
herein.
[0022] FIG. 6 schematically illustrates an example mirror
compatible with certain embodiments described herein.
[0023] FIG. 7 schematically illustrates an example first optical
path of light emitted from the optical fiber in accordance with
certain embodiments described herein.
[0024] FIG. 8 schematically illustrates an example second optical
path of radiation received by the sensor.
[0025] FIG. 9A schematically illustrates an example thermoelectric
element and FIG. 9B schematically illustrates two views of an
example thermal conduit in accordance with certain embodiments
described herein.
[0026] FIG. 10A schematically illustrates another example
thermoelectric element and FIG. 10B schematically illustrates two
views of another example thermal conduit in accordance with certain
embodiments described herein.
[0027] FIG. 11A schematically illustrates a cross-sectional view of
an example heat sink and FIG. 11B schematically illustrates another
example heat sink in accordance with certain embodiments described
herein.
[0028] FIGS. 12A and 12B schematically illustrate two example
configurations of the window with the thermoelectric assembly.
[0029] FIG. 13A schematically illustrates an example chassis for
supporting the various components of the beam delivery apparatus
within the housing in accordance with certain embodiments described
herein.
[0030] FIG. 13B schematically illustrates another example chassis
in accordance with certain embodiments described herein.
[0031] FIG. 14A schematically illustrates a cross-sectional view of
an example configuration of the chassis and the housing in
accordance with certain embodiments described herein.
[0032] FIGS. 14B and 14C schematically illustrate another example
configuration of the chassis and the housing in accordance with
certain embodiments described herein.
[0033] FIGS. 15A and 15B schematically illustrate two states of an
example sensor in accordance with certain embodiments described
herein.
[0034] FIGS. 15C and 15D schematically illustrate two states of
another example sensor in accordance with certain embodiments
described herein.
[0035] FIGS. 16A and 16B schematically illustrate two example
configurations of the trigger force spring and trigger force
adjustment mechanism in accordance with certain embodiments
described herein.
[0036] FIG. 17 schematically illustrates an example lens assembly
sensor in accordance with certain embodiments described herein.
[0037] FIG. 18 is a block diagram of a control circuit comprising a
programmable controller for controlling a light source according to
embodiments described herein.
[0038] FIG. 19A is a graph of the transmittance of light through
blood (in arbitrary units) as a function of wavelength.
[0039] FIG. 19B is a graph of the absorption of light by brain
tissue.
[0040] FIG. 19C shows the efficiency of energy delivery as a
function of wavelength.
[0041] FIG. 20 shows measured absorption of 808 nanometer light
through various rat tissues.
[0042] FIGS. 21A-21D schematically illustrate example pulses in
accordance with certain embodiments described herein.
[0043] FIGS. 22A-22C schematically illustrate an embodiment in
which the apparatus is placed in thermal communication sequentially
with a plurality of treatment sites corresponding to portions of
the patient's scalp.
[0044] FIG. 23 schematically illustrates an example apparatus which
is wearable by a patient for treating the patient's brain.
[0045] FIGS. 24A and 24B schematically illustrate the left-side and
right-side of an example apparatus, respectively, with labels
substantially covering the indicators corresponding to the
treatment sites.
[0046] FIG. 24C schematically illustrates an example labeling
configuration from above a flattened view of the apparatus of FIGS.
24A and 24B.
[0047] FIGS. 25-28 are flow diagrams of example methods for
irradiating a surface with light.
[0048] FIG. 29 is a flow diagram of an example method for
controllably exposing at least one predetermined area of a
patient's scalp to laser light to irradiate the patient's
brain.
[0049] FIG. 30 is a flow diagram of another example method for
treating a patient's brain.
[0050] FIG. 31 is a graph of the power density versus the depth
from the dura for an input power density of 10 mW/cm.sup.2 with the
light bars corresponding to predicted values of the power density
and dark bars corresponding to an estimated minimum working PD of
is 7.5 .mu.W/cm.sup.2, as described below.
[0051] FIG. 32 shows the disposition of patients in the NEST-1
study.
[0052] FIG. 33 shows the mean NIHSS over time for each treatment
group of the NEST-1 study.
[0053] FIG. 34 shows the mean mRS over time for each treatment
group of the NEST-1 study.
[0054] FIG. 35 shows the disposition of patients for the NEST-2
study.
[0055] FIG. 36 shows the distribution of scores on the mRS for the
NEST-2 study.
[0056] FIG. 37 shows the effects of TLT on the primary end point,
on subsets of the data defined by categories of selected baseline
characteristics for the NEST-2 study.
[0057] FIG. 38 is a flow diagram of an example method of treating
brain tissue in accordance with certain embodiments described
herein.
[0058] FIG. 39 is a plot of the neurologic score (NSS) of
control/non-laser-treated (light columns) and laser-irradiated
(dark columns) mice at different time intervals after induction of
brain trauma.
[0059] FIG. 40 shows representative micrographs of cross-sections
of control non-laser-treated and laser-treated mice brains.
[0060] FIG. 41 is a plot of lesion volume of control (light column)
and laser-treated (dark column) traumatized brains 28 days after
induction of head trauma.
[0061] FIGS. 42A-42C show the results of a study of stroke in a
rabbit model using a 6-hour post-embolization time of
treatment.
[0062] FIGS. 43A and 43B show the results of a study of stroke in a
rabbit model using a 12-hour post-embolization time of
treatment.
[0063] FIG. 44 shows the effect of LLT on A.beta. amyloid
deposition in the mouse with mean.+-.SEM for each group. APP
transgenic animals were treated with TLT and no laser, treatments
were administered starting on day 0 (3 months of age) and continued
for 26 weeks (3.times./week treatment). Animals were sacrificed on
the 26.sup.th week and processed to determine the amyloid
burden.
[0064] FIG. 45A shows the effects of LLT on latency time to find
hidden platform (Morris water maze) with mean.+-.SEM for each
group. APP transgenic animals were treated with TLT and no laser,
treatments were administered starting on day 0 (3 months of age)
and continued until day 180 (3.times./week treatment), and were
subjected to water maze and the time to reach the platform was
recorded. Animals were analyzed for day 180 and processed to
determine the latency time.
[0065] FIG. 45B shows the effects of LLT on distance to find hidden
platform (Morris water maze) with mean.+-.SEM for each group. APP
transgenic animals were treated with TLT and no laser, treatments
were administered starting on day 0 (3 months of age) and continued
for 26 weeks (3.times./week treatment), and were subjected to water
maze and the distance to reach the platform was recorded. Animals
were analyzed and processed to determine the distance.
[0066] FIGS. 46A-46C show the effects of LLT on inflammatory
mediators in the brain of APP transgenic mice with mean.+-.SEM for
each group. APP transgenic animals were treated with TLT and no
laser, treatments were administered starting on day 0 (3 months of
age) and continued for 26 weeks (3.times./week treatment). Animals
were sacrificed on the 26.sup.th week and processed to determine
the inflammatory markers, interleukin-1 (IL-1), tumor necrosis
factor-.alpha. (TNF) and transforming growth factor-.beta.
(TGF-.beta.). Animals were analyzed and processed to determine the
inflammatory markers via immunohistochemical and image
analysis.
[0067] FIGS. 47A and 47B show the effects of LLT on A.beta. peptide
levels in the brain of APP transgenic mice with mean.+-.SEM for
each group. APP transgenic animals were treated with TLT and no
laser, treatments were administered starting on day 0 (3 months of
age) and continued for 26 weeks (3.times./week treatment). Animals
were sacrificed on the 26.sup.th week, and the brains were
subjected to A.beta. peptide analysis and compared to no laser
control. Animals were analyzed and processed to determine the brain
A.beta. peptide levels.
[0068] FIGS. 48A and 48B show the effects of LLT on A.beta. peptide
levels in the plasma of APP transgenic mice at 13 and 26 weeks,
respectively, with mean.+-.SEM for each group. APP transgenic
animals were treated with TLT and no laser, treatments were
administered starting on day 0 (3 months of age) and continued for
26 weeks (3.times./week treatment), and the plasma was subjected to
A.beta. peptide analysis and compared to no laser control. Blood
was collected from APP transgenic animals on the 13.sup.th and
26.sup.th week and processed to determine the plasma A.beta.
peptide levels.
[0069] FIGS. 49A and 49B show the effects of LLT on sAPP.alpha. and
CTF.beta. levels, respectively, in the brain of APP transgenic mice
with mean.+-.SEM for each group. APP transgenic animals were
treated with TLT and no laser, treatments were administered
starting on day 0 (3 months of age) and continued for 26 weeks
(3.times./week treatment). Animals were sacrificed on the 26.sup.th
week, and the brain was subjected to sAPP.alpha. and CTF.beta.
protein analysis by western blot and compared to no laser control.
Animals were analyzed and processed to determine the brain protein
levels.
[0070] FIG. 50 shows the effect of LLT on CSF A.beta. peptide
levels in APP transgenic mice with mean.+-.SEM for each group. APP
transgenic animals were treated with TLT and no laser, treatments
were administered starting on day 0 (3 months of age) and continued
for 26 weeks (3.times./week treatment). Animals were sacrificed on
the 26.sup.th week, and the CSF was subjected to total A.beta.
peptide analysis by ELISA and compared to no laser control. Animals
were analyzed and processed to determine the CSF A.beta. peptide
levels.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT
[0071] Low level light therapy ("LLLT") or phototherapy involves
therapeutic administration of light energy to a patient at lower
irradiances than those used for cutting, cauterizing, or ablating
biological tissue, resulting in desirable biostimulatory effects
while leaving tissue undamaged. In non-invasive phototherapy, it is
desirable to apply an efficacious amount of light energy to the
internal tissue to be treated using light sources positioned
outside the body. (See, e.g., U.S. Pat. Nos. 6,537,304 and
6,918,922, both of which are incorporated in their entireties by
reference herein.)
[0072] Laser therapy has been shown to be effective in a variety of
settings, including treating lymphoedema and muscular trauma, and
carpal tunnel syndrome. Recent studies have shown that
laser-generated infrared radiation is able to penetrate various
tissues, including the brain, and to modify function. In addition,
laser-generated infrared radiation can induce effects including,
but not limited to, angiogenesis, modify growth factor
(transforming growth factor-.beta.) signaling pathways, and enhance
protein synthesis.
[0073] However, absorption of the light energy by intervening
tissue can limit the amount of light energy delivered to the target
tissue site, while heating the intervening tissue. In addition,
scattering of the light energy by intervening tissue can limit the
irradiance (or power density) or energy density delivered to the
target tissue site. Brute force attempts to circumvent these
effects by increasing the power and/or irradiance applied to the
outside surface of the body can result in damage (e.g., burning) of
the intervening tissue. For example, a patient experiencing TBI can
have a significant amount of bleeding within the skull (e.g.,
"blood in the field"), and this blood can absorb the applied light,
thereby inhibiting propagation of light energy to brain tissue
below the blood-filled region and heating up.
[0074] Non-invasive phototherapy methods are circumscribed by
setting selected treatment parameters within specified limits so as
to preferably avoid damaging the intervening tissue. A review of
the existing scientific literature in this field would cast doubt
on whether a set of undamaging, yet efficacious, parameters could
be found for treating neurologic conditions. However, certain
embodiments, as described herein, provide devices and methods which
can achieve this goal.
[0075] Such embodiments may include selecting a wavelength of light
at which the absorption by intervening tissue is below a damaging
level. Such embodiments may also include setting the power output
of the light source at low, yet efficacious, irradiances (e.g.,
between approximately 100 .mu.W/cm.sup.2 to approximately 10
W/cm.sup.2) at the target tissue site, setting the temporal profile
of the light applied to the head (e.g. temporal pulsewidths,
temporal pulse shapes, duty cycles, pulse frequencies), and time
periods of application of the light energy at hundreds of
microseconds to minutes to achieve an efficacious energy density at
the target tissue site being treated. Other parameters can also be
varied in the use of phototherapy. These other parameters
contribute to the light energy that is actually delivered to the
treated tissue and may play key roles in the efficacy of
phototherapy. In certain embodiments, the irradiated portion of the
brain can comprise the entire brain.
[0076] In certain embodiments, the target area of the patient's
brain includes the area of injury, e.g., to neurons within the
"zone of danger." In other embodiments, the target area includes
portions of the brain not within the zone of danger. Information
regarding the biomedical mechanisms or reactions involved in
phototherapy is provided by Tiina I. Karu in "Mechanisms of
Low-Power Laser Light Action on Cellular Level", Proceedings of
SPIE Vol. 4159 (2000), Effects of Low-Power Light on Biological
Systems V, Ed. Rachel Lubart, pp. 1-17, and Michael R. Hamblin et
al., "Mechanisms of Low Level Light Therapy," Proc. of SPIE, Vol.
6140, 614001 (2006), each of which is incorporated in its entirety
by reference herein.
[0077] In certain embodiments, the apparatuses and methods of
phototherapy described herein are used to treat physical trauma
(e.g., TBI or ischemic stroke) or other sources of
neurodegeneration. As used herein, the term "neurodegeneration"
refers to the process of cell destruction resulting from primary
destructive events such as stroke or CVA, as well as from
secondary, delayed and progressive destructive mechanisms that are
invoked by cells due to the occurrence of the primary destructive
event. Primary destructive events include disease processes or
physical injury or insult, including stroke, but also include other
diseases and conditions such as multiple sclerosis, amylotrophic
lateral sclerosis, heat stroke, epilepsy, Alzheimer's disease,
dementia resulting from other causes such as AIDS, cerebral
ischemia including focal cerebral ischemia, and physical trauma
such as crush or compression injury in the CNS, including a crush
or compression injury of the brain, spinal cord, nerves or retina,
or any acute injury or insult producing neurodegeneration.
Secondary destructive mechanisms include any mechanism that leads
to the generation and release of neurotoxic molecules, including
but not limited to, apoptosis, depletion of cellular energy stores
because of changes in mitochondrial membrane permeability, release
or failure in the reuptake of excessive glutamate, reperfusion
injury, and activity of cytokines and inflammation. Both primary
and secondary mechanisms contribute to forming a "zone of danger"
for neurons, wherein the neurons in the zone have at least
temporarily survived the primary destructive event, but are at risk
of dying due to processes having delayed effect.
[0078] In certain embodiments, the apparatuses and methods
described herein are used to provide neuroprotection. As used
herein, the term "neuroprotection" refers to a therapeutic strategy
for slowing or preventing the otherwise irreversible loss of
neurons due to neurodegeneration after a primary destructive event,
whether the neurodegeneration loss is due to disease mechanisms
associated with the primary destructive event or secondary
destructive mechanisms.
[0079] In certain embodiments, the apparatuses and methods
described herein are used to improve neurologic function, to
provide neurologic enhancement, or to regain previously lost
neurologic function. The term "neurologic function" as used herein
includes both cognitive function and motor function. The term
"neurologic enhancement" as used herein includes both cognitive
enhancement and motor enhancement. The terms "cognitive
enhancement" and "motor enhancement" as used herein refer to the
improving or heightening of cognitive function and motor function,
respectively.
[0080] The term "cognitive function" as used herein refers to
cognition and cognitive or mental processes or functions, including
those relating to knowing, thinking, learning, perception, memory
(including immediate, recent, or remote memory), and judging.
Symptoms of loss of cognitive function can also include changes in
personality, mood, and behavior of the patient. The term "motor
function" as used herein refers to those bodily functions relating
to muscular movements, primarily conscious muscular movements,
including motor coordination, performance of simple and complex
motor acts, and the like.
[0081] Diseases or conditions affecting neurologic function
include, but are not limited to, Alzheimer's disease, dementia,
AIDS or HIV infection, Cruetzfeldt-Jakob disease, head trauma
(including single-event trauma and long-term trauma such as
multiple concussions or other traumas which may result from
athletic injury), Lewy body disease, Pick's disease, Parkinson's
disease, Huntington's disease, drug or alcohol abuse, brain tumors,
hydrocephalus, kidney or liver disease, stroke, depression, and
other mental diseases which cause disruption in cognitive function,
and neurodegeneration.
Beam Delivery Apparatus
[0082] The phototherapy methods for the treatment of neurologic
conditions (e.g., ischemic stroke, Alzheimer's Disease, Parkinson's
Disease, depression, or TBI) described herein may be practiced and
described using various light delivery systems. Such light delivery
systems may include a low level laser therapy apparatus such as
that shown and described in U.S. Pat. Nos. 6,214,035, 6,267,780,
6,273,905, 6,290,714, and 7,303,578, U.S. patent application Ser.
No. 12/389,294, and U.S. Pat. Appl. Publ. Nos. 2005/0107851 A1 and
2007/0179571 A1, each of which is incorporated in its entirety by
reference herein. For example, in certain embodiments, the light
delivery apparatus can irradiate a portion of the patient's scalp
or skull while cooling the irradiated portion of the scalp or
skull. In certain other embodiments, the irradiated portion of the
patient's scalp or skull is not cooled while irradiating the
portion of the scalp or skull.
[0083] These previously-disclosed light delivery apparatuses were
described primarily in conjunction with phototherapy treatment of
stroke, however in certain embodiments, such light delivery
apparatuses can also be used for phototherapy treatment of other
neurologic conditions (e.g., Alzheimer's Disease, Parkinson's
Disease, depression, TBI). A patient who has experienced a TBI may
have a portion of their scalp damaged, thereby exposing a portion
of their cranium or skull. In certain such embodiments, the light
delivery apparatus can irradiate an exposed portion of the cranium
or skull without the light propagating through scalp tissue.
Certain embodiments described herein are compatible with
irradiation of the brain with light applied to at least a portion
of the scalp or with light applied to at least a portion of the
cranium or skull without propagating through the scalp.
[0084] FIG. 1 schematically illustrates an example beam delivery
apparatus 10 in accordance with certain embodiments described
herein. The apparatus 10 comprises a housing 12, a flexible conduit
14 operatively coupled to the housing 12, and at least one status
indicator 16. In certain embodiments, the apparatus 10 comprises an
output optical assembly 20 comprising an emission surface 22
through which a light beam 30 is emitted. The output optical
assembly 20 is configured to be releasably mechanically coupled to
other components of the apparatus 10.
[0085] In certain embodiments, the housing 12 is sized to be easily
held in one hand (e.g., having a length of approximately 51/2
inches). The housing 12 of certain embodiments further comprises
one or more portions 12a, 12b comprising a biocompatible material
since they may contact the operator, the patient, or both. For
example, one or more low durometer elastomer materials (e.g.
rubber, polymers, thermoplastic resins) can be used in certain
embodiments. The portion 12a is configured to be grasped by a
user's hand during operation of the apparatus 10. The housing 12 of
certain embodiments is configured so that the emission surface 22
can be held in position and sequentially moved by hand to irradiate
selected portions of the patient's skin. In certain embodiments,
the housing 12 comprises one or more recesses or protrusions which
facilitate the housing 12 being gripped by the user. In certain
embodiments, the housing 12 is configured to be placed on a testing
system to measure or monitor the operative parameters of the
apparatus 10. The housing 12 of certain such embodiments comprises
an alignment rib 12c configured to provide a registration
protrusion which mates with a corresponding registration recess on
the testing system to facilitate proper alignment of the emission
surface 22 with the testing system. The housing 12 of certain
embodiments comprises two or more portions (e.g., 2-piece cast
urethane with 60A overmolding or 3-piece Lustran.RTM. with
thermoplastic elastomer overmolding) which fit together to form a
shell in which other operative components are held. In certain
embodiments, the light used by the apparatus 10 can cause eye
damage if viewed by an individual. In such embodiments, the
apparatus 10 can be configured to provide eye protection so as to
avoid viewing of the light by individuals. For example, opaque
materials can be used for the housing 12 and appropriately placed
to block the light from being viewed directly. In addition,
interlocks can be provided so that the light source is not
activated unless the apparatus 10 is in place, or other appropriate
safety measures are taken.
[0086] In certain embodiments, the housing 12 further comprises a
flexible boot 17 generally surrounding the portion of the apparatus
10 which is releasably mounted to the output optical assembly 20.
The boot 17 of certain embodiments provides a barrier to control,
inhibit, prevent, minimize, or reduce contaminants from entering
the housing 12. Thus, by virtue of the boot 17 providing a barrier,
the contamination entering the housing 12 is lower than it would
otherwise be if the boot 17 did not provide a barrier. Example
materials for the flexible boot 17 include but are not limited to,
rubber or another elastomer.
[0087] In certain embodiments, the conduit 14 is configured to
operatively couple the apparatus 10 to various control, power, and
cooling systems that are spaced from the housing 12. In certain
embodiments, the conduit 14 comprises at least one optical fiber
configured to transmit light from a light source to the apparatus
10 to be emitted from the emission surface 22. In certain
embodiments, the conduit 14 further comprises one or more
electrically conductive wires (e.g., one 20-conductor cable, four
6-conductor cables, ground braid) configured to transmit signals
between the apparatus 10 (e.g. trigger switches or temperature
sensors within the apparatus 10) and a control system spaced from
the apparatus 10 and/or to provide electrical power to the
apparatus 10 (e.g. for a thermoelectric cooler) from a power
system. In still other embodiments, the apparatus 10 comprises a
power source (e.g., a battery). In certain embodiments, the conduit
14 comprises one or more coolant tubes (e.g., 0.125-inch inner
diameter) configured to have a coolant (e.g., liquid or gas) flow
to the apparatus 10 from a cooling system. In certain embodiments,
the conduit 14 comprises one or more connectors which are
mechanically coupled to one or more corresponding connectors within
the housing 12. For example, the conduit 14 can comprise an SMA
connector at an end of the optical fiber which is mechanically
coupled to a corresponding SMA mount within the housing 12.
[0088] In certain embodiments, the conduit 14 comprises a
protective sheath around the one or more fibers, wires, and tubes
of the conduit 14. The protective sheath of certain embodiments
controls, inhibits, prevents, minimizes, or reduces light from
exiting the conduit 14 in the event of a failure of the at least
one optical fiber. Thus, by virtue of having the sheath, the light
exiting the conduit 14 upon fiber failure is lower than it would
otherwise be without the sheath. In certain embodiments, the
protective sheath comprises a strain relief apparatus having a
plurality of rigid segments (e.g., stainless steel), with each
segment having a generally cylindrical tubular shape and a
longitudinal axis. Each segment is articulately coupled to
neighboring segments such that an angle between the longitudinal
axes of neighboring segments is limited to be less than a
predetermined angle. In certain embodiments, the protective sheath
allows the conduit 14 to be moved and to bend, but advantageously
limits the radius of curvature of the bend to be sufficiently large
to avoid breaking the one or more fibers, wires, or tubes therein.
In certain embodiments, the sheath comprises a flexible compression
spring (e.g., 4 inches in length) to provide bend relief and/or a
tension line to provide strain relief.
[0089] In certain embodiments, the at least one status indicator 16
comprises one, two, or more light-emitting diodes (LEDs) which are
lit to visually provide the user with information regarding the
status of the apparatus 10. For example, the at least one status
indicator 16 can be used in certain embodiments to indicate when
the laser source is ready to lase pending engagement of the
trigger. In certain embodiments, the LEDs can be lit to show
different colors depending on whether the optical power, electrical
power, or coolant flow being provided to the apparatus 10 are
sufficient for operation of the apparatus 10. In certain
embodiments, the at least one status indicator 16 provides
information regarding whether the output optical assembly 20 is
properly mounted to the apparatus 10. Other types of status
indicators (e.g., flags, sound alarms) are also compatible with
certain embodiments described herein.
[0090] FIG. 2A schematically illustrates a cross-sectional view of
an example output optical assembly 20 in accordance with certain
embodiments described herein. FIG. 2B schematically illustrates
another example output optical assembly 20 in accordance with
certain embodiments described herein. The output optical assembly
20 comprises an optical element 23 comprising the emission surface
22 and a surface 24 facing generally away from the emission surface
22. As used herein, the term "element" is used in its broadest
sense, including, but not limited to, as a reference to a
constituent or distinct part of a composite device. The output
optical assembly 20 further comprises a thermal conduit 25 in
thermal communication with the optical element 23 (e.g., with a
portion of the surface 24). The thermal conduit 25 comprises at
least one surface 26 configured to be in thermal communication with
at least one heat dissipating surface of the apparatus 10 (e.g., a
surface of a cooling mechanism). The output optical assembly 20
further comprises a coupling portion 27 (e.g., spring-loaded 3-pin
bayonet mount or 4-pin bayonet mount) configured to be releasably
attached and detached from the housing 12. In certain embodiments,
the output optical assembly 20 comprises one or more springs which
provide a sufficient force on the at least one surface 26 towards
the at least one heat dissipating surface of the apparatus 10 to
have the desired thermal conductivity between the two. Various
examples of output optical assemblies 20 compatible with certain
embodiments described herein are described more fully in U.S.
patent application Ser. No. 12/233,498, which is incorporated in
its entirety by reference herein.
[0091] In certain embodiments, the output optical assembly 20 is
configured to be placed in thermal communication with the patient's
scalp or skull (e.g., the optical element 23 is configured to
contact the patient's scalp or skull or is configured to be spaced
from the patient's scalp or skull but to contact a thermally
conductive material in contact with the patient's scalp or skull).
In certain embodiments in which the output optical assembly 20 is
cooled, the output optical assembly 20 cools at least a portion of
the patient's scalp or skull (e.g., the portion of the scalp or
skull being irradiated). Thus, in certain embodiments, the output
optical assembly 20 is adapted to control, inhibit, prevent,
minimize, or reduce temperature increases at the scalp or skull
caused by the light. Thus, by virtue of the output optical assembly
20 cooling the portion of the patient's scalp or skull being
irradiated, the temperature of the irradiated portion of the
patient's scalp or skull is lower than it would otherwise be if the
output optical assembly 20 did not cool the irradiated portion of
the scalp or skull. For example, by cooling the irradiated portion
of the patient's scalp or skull using the output optical assembly
20, the temperature of the irradiated portion of the patient's
scalp or skull can be higher than the temperature of the portion of
the patient's scalp or skull if it were not irradiated, but lower
than the temperature of the portion of the patient's scalp or skull
if it were irradiated but not cooled. In certain embodiments, the
patient's scalp comprises hair and skin which cover the patient's
skull. In other embodiments, at least a portion of the hair is
removed prior to the phototherapy treatment, so that the output
optical assembly 20 substantially contacts the skin of the
scalp.
[0092] The optical element 23 of certain embodiments is thermally
conductive, and optically transmissive at wavelengths which are
transmitted by skin. For example, in certain embodiments, the
thermal conductivity of the optical element 23 is sufficient to
remove heat from the irradiated portion of the patient's scalp or
skull, and the optical transmissivity of the optical element 23, at
wavelengths selected to provide the desired irradiance at a target
region of the brain, is sufficient to allow the desired irradiance
of light to propagate through the optical element 23 to irradiate
the patient's scalp or skull. In certain embodiments, the optical
element 23 comprises a rigid material, while in certain other
embodiments, the optical element 23 comprises a low durometer,
thermally conductive, optically transmissive material (e.g., a
flexible bag or container filled with a thermally conductive,
optically transmissive liquid such as water). Example rigid
materials for the optical element 23 include, but are not limited
to, sapphire, diamond, calcium fluoride, and zinc selenide. In
certain embodiments, the optical element 23 has an emission surface
22 configured to face generally towards the surface to be
irradiated (e.g., the patient's scalp or skull). In certain
embodiments, the emission surface 22 is adapted to be placed in
contact with either the irradiated surface or with a substantially
optically transmissive and substantially thermally conductive
material which is in contact with the irradiated surface. The
emission surface 22 of certain embodiments is configured to be in
thermal communication with the surface to be irradiated by the
light beam emitted from the emission surface 22. In certain such
embodiments, the thermal conductivity of the optical element 23 is
sufficiently high to allow heat to flow from the emission surface
22 to the thermal conduit 25 at a sufficient rate to control,
inhibit, prevent, minimize, or reduce damage to the skin or
discomfort to the patient from excessive heating of the skin due to
the irradiation. Thus, by virtue of the thermal conductivity of the
optical element 23, any damage to the skin or discomfort to the
patient can be lower than it would otherwise be if the optical
element 23 did not have a sufficiently high thermal conductivity.
For example, the damage to the skin or discomfort to the patient
can be higher than it would be if the portion of the patient's
scalp were not irradiated, but the damage to the skin or discomfort
to the patient would be lower than it would be if the optical
element 23 did not have a sufficiently high thermal
conductivity.
[0093] In certain embodiments, the optical element 23 has a thermal
conductivity of at least approximately 10 watts/meter-K. In certain
other embodiments, the thermal conductivity of the optical element
23 is at least approximately 15 watts/meter-K. Examples of
materials for the optical element 23 in accordance with certain
embodiments described herein include, but are not limited to,
sapphire which has a thermal conductivity of approximately 23.1
watts/meter-K, and diamond which has a thermal conductivity between
approximately 895 watts/meter-K and approximately 2300
watts/meter-K.
[0094] In certain embodiments, the emission surface 22 is adapted
to conform to the curvature of the scalp or skull. The emission
surface 22 of certain embodiments is concave (e.g., generally
spherical with a radius of curvature of about 100 millimeters). By
fitting to the curvature of the scalp or skull, the emission
surface 22 advantageously controls, inhibits, prevents, minimizes,
or reduces temperature increases at the scalp or skull that would
otherwise result from air-filled gaps between the emission surface
22 and the scalp or skull. Thus, by virtue of the emission surface
22 fitting to the curvature of the portion of the patient's scalp
or skull being irradiated, the temperature of the irradiated
portion of the patient's scalp or skull is lower than it would
otherwise be if the emission surface 22 did not fit to the
curvature of the irradiated portion of the scalp or skull. For
example, by fitting the emission surface 22 to the curvature of the
irradiated portion of the patient's scalp or skull, the temperature
of the irradiated portion of the patient's scalp or skull can be
higher than the temperature of the portion of the patient's scalp
or skull if it were not irradiated, but lower than the temperature
of the portion of the patient's scalp or skull if it were
irradiated but the emission surface 22 did not fit to the portion
of the patient's scalp or skull. The existence of air gaps between
the emission surface 22 and the scalp or skull can reduce the
thermal conductivity between the emission surface 22 and the scalp
or skull, thereby increasing the probability of heating the scalp
or skull by the irradiation.
[0095] In addition, the refractive-index mismatches between such an
air gap and the emission surface 22 and/or the scalp or skull can
cause a portion of the light propagating toward the scalp or skull
to be reflected away from the scalp or skull. In certain
embodiments, the emission surface 22 is placed in contact with the
skin of the scalp or skull so as to advantageously substantially
reduce air gaps between the emission surface 22 and the scalp or
skull in the optical path of the light. In certain other
embodiments in which an intervening material (e.g., a substantially
optically transmissive and substantially thermally conductive gel)
is in contact with the scalp or skull and with the emission surface
22, the emission surface 22 is placed in contact with the
intervening material so as to advantageously avoid creating air
gaps between the emission surface 22 and the intervening material
or between the intervening material and the scalp or skull. In
certain embodiments, the intervening material has a refractive
index at a wavelength of light impinging the scalp which
substantially matches the refractive index of the scalp (e.g.,
about 1.3), thereby reducing any index-mismatch-generated back
reflections between the emission surface 22 and the scalp. Examples
of materials compatible with certain such embodiments described
herein include, but are not limited to, glycerol, water, and silica
gels. Example index-matching gels include, but are not limited to,
those available from Nye Lubricants, Inc. of Fairhaven, Mass.
[0096] In certain embodiments, the emission surface 22 comprises
one or more optical coatings, films, layers, membranes, etc. in the
optical path of the transmitted light which are adapted to reduce
back reflections. By reducing back reflections, the emission
surface 22 increases the amount of light transmitted to the brain
and reduces the need to use higher irradiances which may otherwise
create temperature increases at the scalp or skull.
[0097] In certain embodiments, the output optical assembly 20 is
adapted to diffuse the light prior to reaching the scalp or skull
to advantageously homogenize the light beam prior to reaching the
emission surface 22. Generally, intervening tissues of the scalp
and skull are highly scattering, which can reduce the impact of
non-uniform beam intensity distributions on the illumination of the
patient's cerebral cortex. However, non-uniform beam intensity
distributions with substantial inhomogeneities could result in some
portions of the patient's scalp or skull being heated more than
others (e.g., localized heating where a "hot spot" of the light
beam impinges the patient's scalp or skull). In certain
embodiments, the output optical assembly 20 advantageously
homogenizes the light beam to have a non-uniformity less than
approximately 3 millimeters. FIGS. 3A and 3B schematically
illustrate the diffusive effect on the light by the output optical
assembly 20. An example energy density profile of the light prior
to being transmitted through the output optical assembly 20, as
illustrated by FIG. 3A, is peaked at a particular emission angle.
After being diffused by the output optical assembly 20, as
illustrated by FIG. 3B, the energy density profile of the light
does not have a substantial peak at any particular emission angle,
but is substantially evenly distributed among a range of emission
angles. By diffusing the light, the output optical assembly 20
distributes the light energy substantially evenly over the area to
be illuminated, thereby controlling, inhibiting, preventing,
minimizing, or reducing "hot spots" which would otherwise create
temperature increases at the scalp or skull. Thus, by virtue of the
output optical assembly 20 diffusing the light, the temperature of
the irradiated portion of the patient's scalp or skull is lower
than it would otherwise be if the output optical assembly 20 did
not diffuse the light. For example, by diffusing the light using
the output optical assembly 20, the temperature of the irradiated
portion of the patient's scalp or skull can be higher than the
temperature of the portion of the patient's scalp or skull if it
were not irradiated, but lower than the temperature of the portion
of the patient's scalp or skull if it were irradiated but the light
were not diffused by the output optical assembly 20. In addition,
by diffusing the light prior to reaching the scalp or skull, the
output optical assembly 20 can effectively increase the spot size
of the light impinging the scalp or skull, thereby advantageously
lowering the irradiance at the scalp or skull, as described in U.S.
Pat. No. 7,303,578, which is incorporated in its entirety by
reference herein.
[0098] In certain embodiments, the output optical assembly 20
provides sufficient diffusion of the light such that the irradiance
of the light is less than a maximum tolerable level of the scalp,
skull, or brain. For example, the maximum tolerable level of
certain embodiments is a level at which the patient experiences
discomfort or pain, while in certain other embodiments, the maximum
level is a level at which the patient's scalp or skull is damaged
(e.g., burned). In certain other embodiments, the output optical
assembly 20 provides sufficient diffusion of the light such that
the irradiance of the light equals a therapeutic value at the
subdermal target tissue. The output optical assembly 20 can
comprise example diffusers including, but are not limited to,
holographic diffusers such as those available from Physical Optics
Corp. of Torrance, Calif. and Display Optics P/N SN1333 from
Reflexite Corp. of Avon, Conn.
[0099] In certain embodiments, the output optical assembly 20
provides a reusable interface between the apparatus 10 and the
patient's scalp or skull. In such embodiments, the output optical
assembly 20 can be cleaned or sterilized between uses of the
apparatus 10, particularly between uses by different patients. In
other embodiments, the output optical assembly 20 provides a
disposable and replaceable interface between the apparatus 10 and
the patient's scalp or skull. By using pre-sterilized and
pre-packaged replaceable interfaces, certain embodiments can
advantageously provide sterilized interfaces without undergoing
cleaning or sterilization processing immediately before use.
[0100] In certain embodiments, the output optical assembly 20 is
adapted to apply pressure to at least an irradiated portion of the
scalp. For example, the output optical assembly 20 is capable of
applying pressure to at least an irradiated portion of the scalp
upon a force being applied to the apparatus 10 (e.g., by an
operator of the apparatus 10 pressing the apparatus 10 against the
patient's scalp by hand or by mechanical means to generate force,
such as weights, springs, tension straps). By applying sufficient
pressure, the output optical assembly 20 can blanch the portion of
the scalp by forcing at least some blood out the optical path of
the light energy. (For a general discussion of skin blanching, see,
e.g., A. C. Burton et al., "Relation Between Blood Pressure and
Flow in the Human Forearm," J. Appl. Physiology, Vol. 4, No. 5, pp.
329-339 (1951); A. Matas et al., "Eliminating the Issue of Skin
Color in Assessment of the Blanch Response," Adv. in Skin &
Wound Care, Vol. 14(4, part 1 of 2), pp. 180-188 (July/August
2001); J. Niitsuma et al., "Experimental study of decubitus ulcer
formation in the rabbit ear lobe," J. of Rehab. Res. and Dev., Vol.
40, No. 1, pp. 67-72 (January/February 2003).) The blood removal
resulting from the pressure applied by the output optical assembly
20 to the scalp decreases the corresponding absorption of the light
energy by blood in the scalp. As a result, temperature increases
due to absorption of the light energy by blood at the scalp are
reduced. As a further result, the fraction of the light energy
transmitted to the subdermal target tissue of the brain is
increased. In certain embodiments, a pressure of at least 0.1 pound
per square inch is used to blanch the irradiated portion of the
scalp, while in certain other embodiments, a pressure of at least
one pound per square inch is used to blanch the irradiated portion
of the scalp. In certain embodiments, a pressure of at least about
two pounds per square inch is used to blanch the irradiated portion
of the scalp. Other values or ranges of pressures for blanching the
irradiated portion of the scalp are also compatible with certain
embodiments described herein. The maximum pressure used to blanch
the irradiated portion of the scalp is limited in certain
embodiments by patient comfort levels and tissue damage levels.
[0101] FIGS. 4A and 4B schematically illustrate cross-sectional
views of two example beam delivery apparatuses 10 in accordance
with certain embodiments described herein. In FIGS. 4A and 4B, the
apparatus 10 comprises an output optical assembly 20 having an
emission surface 22 and releasably operatively coupled to the other
components of the apparatus 10. The apparatus 10 comprises an
optical fiber 40, a fiber alignment mechanism 50 operatively
coupled to the optical fiber 40, a mirror 60 in optical
communication with the optical fiber 40, and a window 70 in optical
communication with the mirror 60. During operation of the apparatus
10, light 30 from the optical fiber 40 propagates to the mirror 60
and is reflected by the mirror 60 to propagate through the window
70. The light 30 transmitted through the window 70 propagates
through the output optical assembly 20 along a first optical path
and is emitted from the emission surface 22. In certain
embodiments, the apparatus 10 comprises additional optical elements
(e.g., lenses, diffusers, and/or waveguides) which transmit at
least a portion of the light received via the optical fiber 40 to
the emission surface 22. In certain such embodiments, the
additional optical elements of the apparatus 10 shape, format, or
otherwise modify the light such that the light beam emitted from
the emission surface 22 has the desired beam intensity profile.
[0102] In certain embodiments, the optical fiber 40 comprises a
step-index or graded-index optical fiber. The optical fiber 40 of
certain embodiments is single-mode, while in certain other
embodiments, the optical fiber is multimode. An example optical
fiber 40 compatible with certain embodiments described herein has a
1000-micron diameter and a numerical aperture of approximately
0.22.
[0103] FIG. 5 schematically illustrates an example fiber alignment
mechanism 50 in accordance with certain embodiments described
herein. In certain embodiments, the fiber alignment mechanism 50 is
mechanically coupled to a portion of the optical fiber 40 and is
configured to allow adjustments of the position, tilt, or both of
the end of the optical fiber 40 from which the light is emitted. In
certain embodiments, the fiber alignment mechanism 50 provides an
adjustment range of at least .+-.5 degrees. The fiber alignment
mechanism 50 of FIG. 5 comprises a connector 52 (e.g., SMA
connector) mechanically coupled to the optical fiber 40, a plate 54
(e.g., a kinematic tilt stage) mechanically coupled to the
connector 52, and a plurality of adjustment screws 56 (e.g., 80
turns per inch or 100 turns per inch) adjustably coupled to the
plate 54. By turning the adjustment screws 56, a distance between a
portion of the plate 54 and a corresponding portion of a reference
structure 58 can be adjusted. In certain embodiments, the fiber
alignment mechanism 50 comprises one or more locking screws 59
configured to be tightened so as to fix the plate 54 at a position,
orientation, or both relative to the reference structure 58. Other
configurations of the fiber alignment mechanism 50 are also
compatible with certain embodiments described herein.
[0104] FIG. 6 schematically illustrates an example mirror 60
compatible with certain embodiments described herein. In certain
embodiments, the mirror 60 is substantially reflective of light
emitted from the optical fiber 40 to reflect the light through a
non-zero angle (e.g., 90 degrees). The mirror 60 of certain
embodiments comprises a glass substrate coated on at least one side
by a metal (e.g., gold or aluminum). Examples of mirrors 60
compatible with certain embodiments described herein include, but
are not limited to, a flat, generally planar glass mirror (e.g.,
NT43-886 available from Edmund Optics Inc. of Barrington, N.J.).
The mirror 60 of certain embodiments can be configured to have an
optical power (e.g., the mirror 60 can be concave) and be adapted
to shape, format, or otherwise modify the light to produce a
desired beam intensity profile. In certain embodiments, the mirror
60 is bonded around its perimeter by an adhesive (e.g., OP-29
adhesive available from Dymax Corp. of Torrington, Conn.) to a
support structure 62.
[0105] In certain embodiments, the mirror 60 is partially
transmissive of light emitted from the optical fiber 40. In certain
such embodiments, the support structure 62 comprises an opening and
the apparatus 10 comprises at least one light sensor 64 positioned
to receive light transmitted through the mirror 60 and the opening
of the support structure 62. The at least one light sensor 64 is
configured to generate a signal indicative of the intensity of the
received light, thereby providing a measure of the intensity of the
light reaching the mirror 60. Examples of light sensors 64
compatible with certain embodiments described herein include, but
are not limited to, OPT101 photodiode available from Texas
Instruments of Dallas, Tex. In certain embodiments, a plurality of
light sensors 64 are used to provide operational redundancy to
confirm that light with a sufficient intensity for operation of the
apparatus 10 is being provided by the optical fiber 40. In certain
embodiments, a diffuser 66 is positioned to diffuse the light
transmitted through the mirror 60 before the light impinges the
light sensor 64. In certain embodiments, the light sensor 64 is
protected from stray light by an opaque shroud 68 generally
surrounding the light sensor 64.
[0106] In certain embodiments, the window 70 is substantially
transmissive to infrared radiation. Example windows 70 compatible
with certain embodiments described herein include, but are not
limited to, a flat, generally planar CaF.sub.2 window (e.g.,
TechSpec.RTM. calcium fluoride window available from Edmund Optics
Inc. of Barrington, N.J.).
[0107] In certain embodiments, the window 70 at least partially
bounds a region within the apparatus 10 which contains the mirror
60. The window 70 of certain such embodiments substantially seals
the region against contaminants (e.g., dust, debris) from entering
the region from outside the region. For example, when the output
optical assembly 20 is decoupled from the apparatus 10, the window
70 controls, inhibits, prevents, minimizes, or reduces contaminants
entering the region. Thus, by virtue of the window 70 substantially
sealing the region, the contamination of the region is lower than
it would otherwise be if the window 70 did not substantially seal
the region.
[0108] FIG. 7 schematically illustrates an example first optical
path 32 of light 30 emitted from the optical fiber 40 in accordance
with certain embodiments described herein. The diverging light 30
exiting the optical fiber 40 propagates along the first optical
path 32 towards the mirror 60. The light 30 is reflected by the
mirror 60 and propagates along the first optical path 32 through
the window 70, impinges or is received by the surface 24 of the
optical element 23, and is emitted from the emission surface 22
towards the surface to be irradiated. In certain embodiments, the
mirror 60 reflects the light 30 through an angle of about 90
degrees. In certain embodiments, the mirror 60 is about 2.3 inches
from the face of the optical fiber 40 and the first optical path 32
is about 4.55 inches in length from the fiber output face to the
emission surface 22 of the optical element 23.
[0109] In certain embodiments, the apparatus 10 further comprises a
sensor 80 spaced from the output optical assembly 20. FIG. 8
schematically illustrates an example second optical path 82 of
radiation 84 received by the sensor 80. The sensor 80 is positioned
to receive the radiation 84 from the output optical assembly 20
propagating through the output optical assembly 20 along the second
optical path 82. The first optical path 32 and the second optical
path 82 have a non-zero angle therebetween. In certain embodiments,
the second optical path 82 is co-planar with the first optical path
32, while in certain other embodiments, the first optical path 32
and the second optical path 82 are non-co-planar with one another.
The sensor 80 of certain embodiments receives radiation 84
propagating along the second optical path 82 from at least a
portion of the surface 24 of the optical element 23 during
operation of the apparatus 10.
[0110] The sensor 80 of certain embodiments comprises a temperature
sensor (e.g., thermopile) configured to receive infrared radiation
from a region and to generate a signal indicative of the
temperature of the region. Examples of temperature sensors
compatible with certain embodiments described herein include, but
are not limited to, DX-0496 thermopile available from Dexter
Research Center, Inc. of Dexter, Mich. In certain embodiments, the
field-of-view of the sensor 80 comprises an area of about 0.26
square inches of the surface 24 spaced from the thermal conduit 25
(e.g., by a distance between 0.05 inch and 0.3 inch). In certain
other embodiments, the field-of-view of the sensor 80 comprises an
area of about 0.57 square inches of the surface 24.
[0111] In certain embodiments, the sensor 80 is responsive to the
received radiation 84 by generating a signal indicative of a
temperature of the skin or of a portion of the output optical
assembly 20 (e.g., the optical element 23). In certain such
embodiments, the apparatus 10 further comprises a controller
configured to receive the signal from the sensor 80 and to cause a
warning to be generated, to turn off a source of the light
propagating along the first optical path 32, or both in response to
the signal indicating that the temperature is above a predetermined
threshold temperature (e.g., 42 degrees Celsius).
[0112] The sensor 80 of certain embodiments is not in thermal
communication with the output optical assembly 20. As shown in FIG.
8, the infrared-transmissive window 70 is between the sensor 80 and
the output optical assembly 20. The light 30 propagating along the
first optical path 32 and the infrared radiation 84 propagating
along the second optical path 82 both propagate through the window
70. In certain embodiments, the sensor 80 is wholly or at least
partially within a region of the housing 12 at least partially
bound, and substantially sealed by the window 70 against
contaminants from entering the region from outside the region.
[0113] In certain embodiments, the apparatus 10 is adapted to cool
the irradiated portion of the scalp or skull by removing heat from
the scalp or skull so as to control, inhibit, prevent, minimize, or
reduce temperature increases at the scalp or skull. Thus, by virtue
of the apparatus 10 cooling the irradiated portion of the patient's
scalp or skull, the temperature of the irradiated portion of the
patient's scalp or skull is lower than it would otherwise be if the
apparatus 10 did not cool the irradiated portion of the scalp or
skull. For example, by cooling the irradiated portion of the
patient's scalp or skull using the apparatus 10, the temperature of
the irradiated portion of the patient's scalp or skull can be
higher than the temperature of the portion of the patient's scalp
or skull if it were not irradiated, but lower than the temperature
of the portion of the patient's scalp or skull if it were
irradiated but not cooled. Referring to FIGS. 4A and 4B, in certain
embodiments, the apparatus 10 comprises a thermoelectric assembly
90 and a heat sink 100 in thermal communication with the
thermoelectric assembly 90. In certain embodiments, the
thermoelectric assembly 90 actively cools the patient's scalp or
skull via the output optical assembly 20, thereby advantageously
avoiding large temperature gradients at the patient's scalp or
skull which would otherwise cause discomfort to the patient. In
certain embodiments, the apparatus 10 further comprises one or more
temperature sensors (e.g., thermocouples, thermistors) which
generate electrical signals indicative of the temperature of the
thermoelectric assembly 90.
[0114] In certain embodiments, the thermoelectric assembly 90
comprises at least one thermoelectric element 91 and a thermal
conduit 92. The at least one thermoelectric element 91 of the
thermoelectric assembly 90 is responsive to an electric current
applied to the thermoelectric assembly 90 by cooling at least a
first surface 93 of the thermoelectric assembly 90 and heating at
least a second surface 94 of the thermoelectric assembly 90. The
thermoelectric assembly 90 is configured to be releasably
mechanically coupled to the output optical assembly 20 so as to
have the first surface 93 in thermal communication with the output
optical assembly 20. In certain embodiments, the first surface 93
comprises a surface of the thermal conduit 92 and the second
surface 94 comprises a surface of the thermoelectric element
91.
[0115] FIG. 9A schematically illustrates an example thermoelectric
element 91 and FIG. 9B schematically illustrates two views of an
example thermal conduit 92 in accordance with certain embodiments
described herein. FIG. 10A schematically illustrates another
example thermoelectric element 91 and FIG. 10B schematically
illustrates two views of another example thermal conduit 92 in
accordance with certain embodiments described herein. The
thermoelectric element 91 has a surface 95 configured to be in
thermal communication with a corresponding surface 96 of the
thermal conduit 92 (e.g., by a thermally conductive adhesive). Upon
application of an electric current to the thermoelectric element
91, the second surface 94 is heated and the surface 95 is cooled,
thereby cooling the first surface 93. In certain such embodiments,
the first surface 93 serves as at least one heat dissipating
surface of the apparatus 10 configured to be in thermal
communication with the at least one surface 26 of the thermal
conduit 25 of the output optical assembly 20 (e.g., by contacting
or mating so as to provide a thermally conductive connection
between the thermoelectric assembly 26 and the output optical
assembly 20). By having the thermally conductive output optical
assembly 20 in thermal communication with the thermoelectric
assembly 90, certain embodiments advantageously provide a conduit
for heat conduction away from the treatment site (e.g. the skin).
In certain embodiments, the output optical assembly 20 is pressed
against the patient's skin and transfers heat away from the
treatment site.
[0116] Examples of thermoelectric elements 91 compatible with
certain embodiments described herein include, but are not limited
to, DT12-6, Q.sub.max=60 W, square thermoelectric element available
from Marlow Industries of Dallas, Tex., and Q.sub.max=45 W
toroidal- or donut-shaped thermoelectric element from Ferrotec
Corp. of Bedford, N.H. In certain embodiments, the thermoelectric
element 91 removes heat from the output optical assembly 20 at a
rate in a range of about 0.1 Watt to about 5 Watts or in a range of
about 1 Watt to about 3 Watts. Example temperature controllers for
operating the thermoelectric assembly 90 in accordance with certain
embodiments described herein include, but are not limited to,
MPT-5000 available from Wavelength Electronics, Inc. of Bozeman,
Mont. Example materials for the thermal conduit 92 compatible with
certain embodiments described herein include, but are not limited
to, aluminum and copper. The thermal conduit 92 of certain
embodiments has a thermal mass in a range of about 30 grams to
about 70 grams, and has a thermal length between surface 93 and
surface 96 in a range of about 0.5 inch to about 3.5 inches.
[0117] In certain embodiments, the thermoelectric assembly 90
generally surrounds a first region 97, wherein, during operation of
the apparatus 10, light irradiating a portion of the patient's skin
propagates through the first region 97. As shown in FIGS. 9B and
10B, in certain embodiments, the first region 97 comprises an
aperture through the thermal conduit 92. As shown in FIG. 10B, the
first region 97 in certain embodiments further comprises an
aperture through the thermoelectric element 91. In certain
embodiments, the thermoelectric assembly 90 comprises a plurality
of thermoelectric elements 91 which are spaced from one another and
are distributed to generally surround the first region 97. As used
herein, the term "generally surrounds" has its broadest reasonable
interpretation, including but not limited to, encircles or extends
around at least one margin of the region, or being distributed
around at least one margin of the region with one or more gaps
along the at least one margin.
[0118] FIG. 11A schematically illustrates a cross-sectional view of
an example heat sink 100 and FIG. 11B schematically illustrates
another example heat sink 100 in accordance with certain
embodiments described herein. The heat sink 100 comprises an inlet
101, an outlet 102, and a fluid conduit 103 in fluid communication
with the inlet 101 and the outlet 102. The inlet 101 and the outlet
102 of certain embodiments comprise stainless steel barbs
configured to be connected to tubes (e.g., using nylon or stainless
steel hose barb locks, clamps, or crimps) which provide a coolant
(e.g., water, air, glycerol) to flow through the fluid conduit 103
and to remove heat from the fluid conduit 103. In certain
embodiments, the coolant is provided by a chiller or other heat
transfer device which cools the coolant prior to its being supplied
to the heat sink 100.
[0119] The example heat sink 100 of FIG. 11A is machined from an
aluminum block and has a recess 104 in which the thermoelectric
assembly 90 is placed to provide thermal communication between the
heat sink 100 and the second surface 94 of the thermoelectric
assembly 90. The example heat sink 100 of FIG. 11B comprises a
first portion 105 and a second portion 106 which fit together to
form the coolant conduit 103. In certain embodiments, a thermally
conductive adhesive (e.g., EP1200 thermal adhesive available from
Resinlab, LLC of Germantown, Wis., with a 0.005-inch stainless
steel wire to set the bondline) is used to bond the thermoelectric
assembly 90 and the heat sink 100 together in thermal communication
with one another.
[0120] The output optical assembly 20 comprises a thermally
conductive thermal conduit 25 having at least one surface 26
configured to be in thermal communication with the first surface of
the thermoelectric assembly 90. As shown in FIGS. 2A and 2B, the
thermal conduit 25 generally surrounds a second region 28. During
operation of the apparatus 10, the light propagates through the
first region 97, the second region 28, and the optical element 23.
In certain embodiments, the heat sink 100 generally surrounds a
third region 107, as schematically illustrated by FIG. 1l B. During
operation of the apparatus 10 in certain such embodiments, the
light propagates through the third region 107, the first region 97,
the second region 28, and the optical element 23.
[0121] FIGS. 12A and 12B schematically illustrate two example
configurations of the window 70 with the thermoelectric assembly
90. In certain embodiments, the window 70 is in thermal
communication with at least a portion of the thermoelectric
assembly 90 (e.g., bonded to a recess in the thermal conduit 92, as
shown in FIG. 12A, using OP-29 adhesive available from Dymax Corp.
of Torrington, Conn.). In certain embodiments, the window 70 is in
thermal communication with at least a portion of the heat sink 100
(e.g., retained by an o-ring in the heat sink 100), as shown in
FIG. 12B. In certain embodiments, the window 70 is not in thermal
communication with either the thermoelectric assembly 90 or the
heat sink 100.
[0122] FIG. 13A schematically illustrates an example chassis 110
for supporting the various components of the beam delivery
apparatus 10 within the housing 12 in accordance with certain
embodiments described herein. The chassis 110 of FIG. 13A comprises
a single unitary or monolithic piece which is machined to provide
various surfaces and holes used to mount the various components of
the beam delivery apparatus 10. FIG. 13B schematically illustrates
another example chassis 110 in accordance with certain embodiments
described herein. The chassis 110 of FIG. 13B comprises a plurality
of portions which are bolted or pinned together.
[0123] FIG. 14A schematically illustrates a cross-sectional view of
an example configuration of the chassis 110 and the housing 12 in
accordance with certain embodiments described herein. The chassis
110 of certain embodiments is electrically connected to ground,
while in certain other embodiments, the chassis 110 is electrically
insulated from ground (e.g., floating). In certain embodiments, the
chassis 110 is configured to move relative to the housing 12. For
example, the chassis 110 and the housing 12 are mechanically
coupled together by a pivot 112, as schematically illustrated by
FIG. 14A. The optical fiber 40, fiber adjustment apparatus 50,
mirror 60, window 70, sensor 80, and heat sink 100 are each
mechanically coupled to the chassis 110. The output optical
assembly 20 is also mechanically coupled to the chassis 110 via the
thermoelectric assembly 90 and the heat sink 100.
[0124] For the configuration of FIG. 14A, the emission surface 22
of the output optical assembly 20 is placed in thermal
communication (e.g., in contact) with the patient's scalp or skull
by a user pressing the housing 12 towards the scalp or skull. The
pivot 112 allows the chassis 110 to rotate about the pivot 112
relative to the housing 12 (e.g., by an angle between 1 and 2
degrees, or about 1.75 degrees) such that the emission surface 22
moves towards the housing 12 (e.g., by a distance of 0.05-0.3 inch,
or about 0.1 inch). In certain such embodiments, this movement of
the chassis 110, as well as of the fiber adjustment apparatus 50
and the optical fiber 40, results in a flexing of a portion of the
optical fiber 40 (e.g., in proximity to the coupling between the
housing 12 and the conduit 14).
[0125] This flexing of the optical fiber 40 can be undesirable in
certain circumstances, such as when the optical fiber 40 or its
connection to the fiber adjustment apparatus 50 is fragile and
prone to breakage or failure due to repeated flexing. FIGS. 14B and
14C schematically illustrate another example configuration of the
chassis 110 and the housing 12 in accordance with certain
embodiments described herein. The chassis 110 comprises a first
chassis element 120 and a second chassis element 122 mechanically
coupled to the first chassis element 120 such that the first
chassis element 120 and the second chassis element 122 can move
relative to one another. For example, in certain embodiments, the
apparatus 10 further comprises a hinge 124 (e.g., a pivot or
flexible portion) about which the first chassis element 120 and the
second chassis element 122 are configured to deflect relative to
one another.
[0126] In certain embodiments, the first chassis element 120 is
mechanically coupled to the housing 12, and the optical fiber 40,
fiber adjustment apparatus 50, mirror 60, and sensor 80 (each shown
in dotted lines in FIG. 14C) are mechanically coupled to the first
chassis element 120. The second chassis element 122 is mechanically
coupled to the window 70, thermoelectric assembly 90, and the heat
sink 100 (each shown in dotted lines in FIG. 14C). The output
optical assembly 20 is also mechanically coupled to the second
chassis element 122 via the thermoelectric assembly 90 and the heat
sink 100. Thus, in certain such embodiments, a first portion of the
apparatus 10 comprises the housing 12, first chassis element 120,
optical fiber 40, fiber adjustment apparatus 50, mirror 60, and
sensor 80, and a second portion of the apparatus 10 comprises the
second chassis element 122, window 70, thermoelectric assembly 90,
heat sink 100, and output optical assembly 20. The second portion
is mechanically coupled to the first portion and is in optical
communication with the first portion. The second portion is
configured to be placed in thermal communication with the patient's
skin such that the light from the first portion propagates through
the second portion during operation of the apparatus 10. The first
portion and the second portion are configured to move relative to
one another in response to the second portion being placed in
thermal communication with the patient's skin.
[0127] In certain embodiments, the second portion comprises the
output optical assembly 20 and the first portion and the second
portion are configured to deflect relative to one another by a
non-zero angle. In certain embodiments, this deflection occurs upon
the output optical assembly 20 applying a pressure to a portion of
the patient's scalp sufficient to at least partially blanch the
portion of the patient's scalp. In certain embodiments, this
deflection occurs upon the output optical assembly 20 being placed
in thermal communication with the patient's scalp or skull. In
certain embodiments, the apparatus 10 further comprises a spring
mechanically coupled to the first portion and the second portion.
The spring provides a restoring force in response to movement of
the first portion and the second portion relative to one
another.
[0128] For the configuration of FIGS. 14B and 14C, the emission
surface 22 of the output optical assembly 20 is placed in thermal
communication (e.g., in contact) with the patient's scalp or skull
by a user pressing the housing 12 towards the scalp or skull. The
hinge 124 allows the second portion (e.g., including the second
chassis element 122) to rotate about the hinge 124 relative to the
first portion (e.g., including the first chassis element 120). This
rotation can be by an angle between 1 and 3 degrees, or about 2.3
degrees) such that the emission surface 22 moves towards the
housing 12 (e.g., by a distance of 0.05-0.3 inch, or about 0.08
inch). In certain such embodiments in which the first portion
comprises the optical fiber 40, deflection of the first portion and
the second portion relative to one another controls, inhibits,
prevents, minimizes, or reduces flexing or movement of the optical
fiber 40 (e.g., to control, inhibit, prevent, minimize, or reduce
damage to the optical fiber 40). Thus, by virtue of the movement of
the first and second portions relative to one another, the flexing,
movement, or damage of the optical fiber 40 is lower than it would
otherwise be if the first and second portions did not move relative
to one another.
[0129] In certain embodiments, the relative movement of the output
optical assembly 20 and the mirror 60 can result in the light beam
30 being at least partially occluded or "clipped" by the thermal
conduit 25 of the output optical assembly 20. For example, for a
light beam diameter of 30 millimeters, the light beam 30 is not
clipped by the thermal conduit 25. For larger light beam diameters,
the light beam 30 is partially occluded by the thermal conduit 25.
For a light beam diameter of 31 millimeters, about 0.02% of the
light beam area is occluded, and for 32 millimeters, about 1.56% of
the light beam area is occluded, resulting in an estimated power
loss of less than about 0.08%.
[0130] In certain embodiments, the apparatus 10 further comprises a
sensor 130 configured to detect movement of the first portion and
the second portion relative to one another (e.g., movement of the
first chassis element 120 and the second chassis element 122
relative to one another). The sensor 130 is configured to transmit
a signal to a controller configured to receive the signal and to
control a light source in response to the signal, where the light
source is configured to generate the light used by the apparatus 10
irradiate the patient's scalp or skull. In certain embodiments, the
sensor 130 transmits the signal to the controller upon detecting
that the movement between the first portion and the second portion
is larger than a predetermined threshold value. In this way, the
sensor 130 serves as a trigger switch which is used to trigger the
apparatus 10 (e.g., providing the apparatus 10 with light upon the
sensor 130 detecting the predetermined amount of movement between
the first portion and the second portion indicative of the
apparatus 10 being in a condition for use). The trigger switch of
certain embodiments is actuated by pressing the output optical
assembly 20 against a surface. The light source providing light to
the apparatus 10 is responsive to the trigger switch by emitting
light only when the trigger switch is actuated. Therefore, in
certain such embodiments, to utilize the apparatus 10, the output
optical assembly 20 is pressed against the patient's skin, such as
described above.
[0131] FIGS. 15A and 15B schematically illustrate two states of an
example sensor 130 in accordance with certain embodiments described
herein. The sensor 130 comprises at least one trigger flag 132
mechanically coupled to the first portion (e.g., the housing 12)
and at least one optical switch 134 mechanically coupled to the
second portion (e.g., the second chassis element 122). For example,
the at least one optical switch 134 of certain embodiments
comprises one, two, or more EE-SX-1035 optical switches available
from Omron Electronics Components LLC of Schaumburg, Ill. In a
first state, the trigger flag 132 is displaced away from a sensor
light beam which is detected by the optical switch 134. Upon
pressing the output optical assembly 20 in thermal communication
with the patient's scalp or skull, the optical switch 134 moves
relative to the trigger flag 132 (e.g., by a distance of about 0.07
inch) such that the trigger flag 132 intercepts the sensor light
beam such that it is no longer detected by the optical switch 134.
In response to this second state, the sensor 130 generates a
corresponding signal. In certain other embodiments, the trigger
flag 132 can be positioned to intercept the sensor light beam in
the first state and to not intercept the sensor light beam in the
second state.
[0132] FIGS. 15C and 15D schematically illustrate two states of
another example sensor 130 in accordance with certain embodiments
described herein. The sensor 130 comprises a reflective element 135
mechanically coupled to the first portion (e.g., the first chassis
element 120) and at least one light source/detector pair 136
mechanically coupled to the second portion (e.g., the second
chassis element 122). For example, the at least one light
source/detector pair 136a, 136b of certain embodiments comprises
one, two, or more QRE1113GR reflective sensors available from
Fairchild Semiconductor Corp. of San Jose, Calif. In a first state,
the reflective surface 135 is a first distance away from the light
source/detector pair 136a, 136b such that a sensor light beam from
the source 136a is reflected from the surface 135 but is not
detected by the detector 136b. Upon pressing the output optical
assembly 20 in thermal communication with the patient's scalp or
skull, the reflective surface 135 moves (e.g., by a distance of
about 0.04 inch) to be a second distance away from the light
source/detector pair 136a, 136b such that the sensor light beam
from the source 136a is reflected from the surface 135 and is
detected by the detector 136b. In response to this second state,
the sensor 130 generates a corresponding signal. In certain
embodiments, the sensor 130 further comprises a shroud 137
configured to protect the detector 136b from stray light. In
certain other embodiments, the reflective surface 135 can be
positioned to reflect the sensor light beam to the detector 136b in
the first state and to not reflect the sensor light beam to the
detector 136b in the second state.
[0133] In certain embodiments, the apparatus 10 further comprises
an adjustment mechanism configured to set the predetermined
threshold value, to change the predetermined threshold value, or
both. In certain such embodiments, the adjustment mechanism
comprises a set screw which changes the relative positions of the
two portions of the sensor 130 which move relative to one another.
Certain embodiments further comprise a stop configured to limit a
range of movement of the first portion and the second portion
relative to one another.
[0134] In certain embodiments, the apparatus 10 comprises a trigger
force spring 140 and a trigger force adjustment mechanism 142.
FIGS. 16A and 16B schematically illustrate two example
configurations of the trigger force spring 140 and trigger force
adjustment mechanism 142 in accordance with certain embodiments
described herein. The trigger force spring 140 is mechanically
coupled to the first portion (e.g., the first chassis element 120)
and the second portion (e.g., the second chassis element 122) and
provides a restoring force when the first portion and the second
portion are moved relative to one another. The trigger force
adjustment mechanism 142 of FIG. 16A comprises one or more shims
(e.g., each shim providing about 100 grams of adjustment) placed
between the spring 140 and at least one of the first portion and
the second portion. The trigger force adjustment mechanism 142 of
FIG. 16B comprises one, two, or more adjustment set screws. In
either configuration, the trigger force adjustment mechanism 142
compresses the spring 140 to adjust the amount of force which will
move the first and second portions relative to one another by a
sufficient amount to trigger the apparatus 10. In certain
embodiments, the trigger force adjustment mechanism 142 is set such
that the apparatus 10 is triggered by a pressure applied to the
emission surface 22 towards the housing 12 of at least 0.1 pound
per square inch, at least one pound per square inch, or at least
about two pounds per square inch.
[0135] In certain embodiments, the apparatus 10 further comprises a
lens assembly sensor 150 configured to detect the presence of the
output optical assembly 20 mounted on the apparatus 10. FIG. 17
schematically illustrates an example lens assembly sensor 150 in
accordance with certain embodiments described herein. For example,
the lens assembly sensor 150 of certain embodiments comprises at
least one reflective surface 152 and at least one light
source/detector pair 154a, 154b (e.g., one, two, or more QRE1113GR
reflective sensors available from Fairchild Semiconductor Corp. of
San Jose, Calif.). The reflective surface 152 moves relative to the
light source/detector pair 154a, 154b upon mounting the output
optical assembly 20 to be in thermal communication with the thermal
conduit 92. For example, when the output optical assembly 20 is
mounted, the bayonet is pulled downward. In response to this
movement, the sensor 150 generates a corresponding signal. In
certain embodiments, the sensor 150 further comprises a shroud 156
configured to protect the detector 154b from stray light.
Control Circuit
[0136] FIG. 18 is a block diagram of a control circuit 200
comprising a programmable controller 205 for controlling a light
source 207 according to embodiments described herein. The control
circuit 200 is configured to adjust the power of the light energy
generated by the light source 207 such that the light emitted from
the emission surface 22 generates a predetermined surface
irradiance at the scalp or skull corresponding to a predetermined
energy delivery profile, such as a predetermined subsurface
irradiance, to the target area of the brain.
[0137] In certain embodiments, the programmable controller 205
comprises a logic circuit 210, a clock 212 coupled to the logic
circuit 210, and an interface 214 coupled to the logic circuit 210.
The clock 212 of certain embodiments provides a timing signal to
the logic circuit 210 so that the logic circuit 210 can monitor and
control timing intervals of the applied light. Examples of timing
intervals include, but are not limited to, total treatment times,
pulsewidth times for pulses of applied light, and time intervals
between pulses of applied light. In certain embodiments, the light
source 207 can be selectively turned on and off to reduce the
thermal load on the scalp or skull and to deliver a selected
irradiance to particular areas of the brain.
[0138] The interface 214 of certain embodiments provides signals to
the logic circuit 210 which the logic circuit 210 uses to control
the applied light. The interface 214 can comprise a user interface
or an interface to a sensor monitoring at least one parameter of
the treatment. In certain such embodiments, the programmable
controller 126 is responsive to signals from the sensor to
preferably adjust the treatment parameters to optimize the measured
response. The programmable controller 126 can thus provide
closed-loop monitoring and adjustment of various treatment
parameters to optimize the phototherapy. The signals provided by
the interface 214 from a user are indicative of parameters that may
include, but are not limited to, patient characteristics (e.g.,
skin type, fat percentage), selected applied irradiances, target
time intervals, and irradiance/timing profiles for the applied
light.
[0139] In certain embodiments, the logic circuit 210 is coupled to
a light source driver 220. The light source driver 220 is coupled
to a power supply 230, which in certain embodiments comprises a
battery and in other embodiments comprises an alternating current
source. The light source driver 220 is also coupled to the light
source 207. The logic circuit 210 is responsive to the signal from
the clock 212 and to user input from the user interface 214 to
transmit a control signal to the light source driver 220. In
response to the control signal from the logic circuit 210, the
light source driver 220 adjust and controls the power applied to
the light source. Other control circuits besides the control
circuit 200 of FIG. 18 are compatible with embodiments described
herein.
[0140] In certain embodiments, the logic circuit 110 is responsive
to signals from a sensor monitoring at least one parameter of the
treatment to control the applied light. For example, certain
embodiments comprise a temperature sensor in thermal communication
with the scalp or skull to provide information regarding the
temperature of the scalp or skull to the logic circuit 210. In such
embodiments, the logic circuit 210 is responsive to the information
from the temperature sensor to transmit a control signal to the
light source driver 220 so as to adjust the parameters of the
applied light to maintain the scalp or skull temperature below a
predetermined level. Other embodiments include example biomedical
sensors including, but not limited to, a blood flow sensor, a blood
gas (e.g., oxygenation) sensor, an ATP production sensor, or a
cellular activity sensor. Such biomedical sensors can provide
real-time feedback information to the logic circuit 210. In certain
such embodiments, the logic circuit 110 is responsive to signals
from the sensors to preferably adjust the parameters of the applied
light to optimize the measured response. The logic circuit 110 can
thus provide closed-loop monitoring and adjustment of various
parameters of the applied light to optimize the phototherapy.
Light Parameters
[0141] The various parameters of the light beam emitted from the
emission surface 22 are advantageously selected to provide
treatment while controlling, inhibiting, preventing, minimizing, or
reducing injury or discomfort to the patient due to heating of the
scalp or skull by the light. While discussed separately, these
various parameters below can be combined with one another within
the disclosed values in accordance with embodiments described
herein.
Wavelength
[0142] In certain embodiments, light in the visible to
near-infrared wavelength range is used to irradiate the patient's
scalp or skull. In certain embodiments, the light is substantially
monochromatic (i.e., light having one wavelength, or light having a
narrow band of wavelengths). So that the amount of light
transmitted to the brain is maximized, the wavelength of the light
is selected in certain embodiments to be at or near a transmission
peak (or at or near an absorption minimum) for the intervening
tissue. In certain such embodiments, the wavelength corresponds to
a peak in the transmission spectrum of tissue at about 820
nanometers. In certain other embodiments, the light comprises one
or more wavelengths between about 630 nanometers and about 1064
nanometers, between about 600 nanometers and about 980 nanometers,
between about 780 nanometers and about 840 nanometers, between
about 805 nanometers and about 820 nanometers, or includes
wavelengths of about 785, 790, 795, 800, 805, 810, 815, 820, 825,
or 830 nanometers. An intermediate wavelength in a range between
approximately 730 nanometers and approximately 750 nanometers
(e.g., about 739 nanometers) appears to be suitable for penetrating
the skull, although other wavelengths are also suitable and may be
used. In other embodiments, a plurality of wavelengths is used
(e.g. applied concurrently or sequentially). In certain
embodiments, the light has a wavelength distribution peaked at a
peak wavelength and has a linewidth less than .+-.10 nanometers
from the peak wavelength. In certain such embodiments, the light
has a linewidth less than 4 nanometers, full width at 90% of
energy. In certain embodiments, the center wavelength is
(808.+-.10) nanometers with a spectral linewidth less than 4
nanometers, full width at 90% of energy.
[0143] In certain embodiments, the light is generated by a light
source comprising one or more laser diodes, which each provide
coherent light. In embodiments in which the light from the light
source is coherent, the emitted light may produce "speckling" due
to coherent interference of the light. This speckling comprises
intensity spikes which are created by wavefront interference
effects and can occur in proximity to the target tissue being
treated. For example, while the average irradiance or power density
may be approximately 10 mW/cm.sup.2, the power density of one such
intensity spike in proximity to the brain tissue to be treated may
be approximately 300 mW/cm.sup.2. In certain embodiments, this
increased power density due to speckling can improve the efficacy
of treatments using coherent light over those using incoherent
light for illumination of deeper tissues. In addition, the
speckling can provide the increased power density without
overheating the tissue being irradiated. The light within the
speckle fields or islands containing these intensity spikes is
polarized, and in certain embodiments, this polarized light
provides enhanced efficacy beyond that for unpolarized light of the
same intensity or irradiance.
[0144] In certain embodiments, the light source includes at least
one continuously emitting GaAlAs laser diode having a wavelength of
about 830 nanometers. In another embodiment, the light source
comprises a laser source having a wavelength of about 808
nanometers. In still other embodiments, the light source includes
at least one vertical cavity surface-emitting laser (VCSEL) diode.
Other light sources compatible with embodiments described herein
include, but are not limited to, light-emitting diodes (LEDs) and
filtered lamps.
[0145] In certain embodiments, the one or more wavelengths are
selected so as to work with one or more chromophores within the
target tissue. Without being bound by theory or by a specific
mechanism, it is believed that irradiation of chromophores
increases the production of ATP in the target tissue and/or
controls, inhibits, prevents, minimizes, or reduces apoptosis of
the injured tissues, thereby producing beneficial effects, as
described more fully below.
[0146] Some chromophores, such as water or hemoglobin, are
ubiquitous and absorb light to such a degree that little or no
penetration of light energy into a tissue occurs. For example,
water absorbs light above approximately 1300 nanometers. Thus
energy in this range has little ability to penetrate tissue due to
the water content. However, water is transparent or nearly
transparent in wavelengths between 300 and 1300 nanometers. Another
example is hemoglobin, which absorbs heavily in the region between
300 and 670 nanometers, but is reasonably transparent above 670
nanometers.
[0147] Based on these broad assumptions, one can define an "IR
window" into the body. Within the window, there are certain
wavelengths that are more or less likely to penetrate. This
discussion does not include wavelength dependent scattering effects
of intervening tissues.
[0148] The absorption/transmittance of various tissues have been
directly measured to determine the utility of various wavelengths.
FIG. 19A is a graph of the transmittance of light through blood (in
arbitrary units) as a function of wavelength. Blood absorbs less in
the region above 700 nanometers, and is particularly transparent at
wavelengths above 780 nanometers. Wavelengths below 700 nanometers
are heavily absorbed, and are not likely to be useful
therapeutically (except for topical indications).
[0149] FIG. 19B is a graph of the absorption of light by brain
tissue. Absorption in the brain is strong for wavelengths between
620 and 980 nanometers. This range is also where the copper centers
in mitochondria absorb. The brain is particularly rich in
mitochondria as it is a very active tissue metabolically (the brain
accounts for 20% of blood flow and oxygen consumption). As such,
the absorption of light in the 620 to 980 nanometer range is
expected if a photostimulative effect is to take place.
[0150] By combining FIGS. 19A and 19B, the efficiency of energy
delivery as a function of wavelength can be calculated, as shown in
FIG. 19C. Wavelengths between 780 and 880 nanometers are preferable
(efficiency of 0.6 or greater) for targeting the brain. The peak
efficiency is about 800 to 830 nanometers (efficiency of 1.0 or
greater). These wavelengths are not absorbed by water or
hemoglobin, and are likely to penetrate to the brain. Once these
wavelengths reach the brain, they will be absorbed by the brain and
converted to useful energy.
[0151] These effects have been directly demonstrated in rat
tissues. The absorption of 808 nanometer light was measured through
various rat tissues, as shown in FIG. 20. Soft tissues such as skin
and fat absorb little light. Muscle, richer in mitochondria,
absorbs more light. Even bone is fairly transparent. However, as
noted above, brain tissue, as well as spinal cord tissue, absorb
808 nanometer light well.
Irradiance or Power Density
[0152] In certain embodiments, the light beam has a time-averaged
irradiance or power density at the emission surface 22 of the
output optical assembly 20 between about 10 mW/cm.sup.2 to about 10
W/cm.sup.2, between about 100 mW/cm.sup.2 to about 1000
mW/cm.sup.2, between about 500 mW/cm.sup.2 to about 1 W/cm.sup.2,
or between about 650 mW/cm.sup.2 to about 750 mW/cm.sup.2 across
the cross-sectional area of the light beam. For a pulsed light
beam, the time-averaged irradiance is averaged over a time period
long compared to the temporal pulse widths of the pulses (e.g.,
averaged over a fraction of a second longer than the temporal pulse
width, over 1 second, or over multiple seconds). For a
continuous-wave (CW) light beam with time-varying irradiance, the
time-averaged irradiance can be an average of the instantaneous
irradiance averaged over a time period longer than a characteristic
time period of fluctuations of the light beam. In certain
embodiments, a duty cycle in a range between 1% and 80%, between
10% and 30%, or about 20% can be used with a peak irradiance at the
emission surface 22 of the output optical assembly 20 between about
12.5 mW/cm.sup.2 to about 1000 W/cm.sup.2, between about 50
mW/cm.sup.2 to about 50 W/cm.sup.2, between about 500 mW/cm.sup.2
to about 5000 mW/cm.sup.2, between about 2500 mW/cm.sup.2 to about
5 W/cm.sup.2, or between about 3.25 W/cm.sup.2 to about 3.75
W/cm.sup.2 across the cross-sectional area of the light beam. In
certain embodiments, the pulsed light beam has an energy or fluence
(e.g., peak irradiance multiplied by the temporal pulsewidth) at
the emission surface 22 of the output optical assembly 20 between
about 12.5 .mu.J/cm.sup.2 to about 1 J/cm.sup.2, between about 50
.mu.J/cm.sup.2 to about 50 mJ/cm.sup.2, between about 500
.mu.J/cm.sup.2 to about 5 mJ/cm.sup.2, between about 2.5
mJ/cm.sup.2 to about 5 mJ/cm.sup.2, or between about 3.25
mJ/cm.sup.2 to about 3.75 mJ/cm.sup.2.
[0153] The cross-sectional area of the light beam of certain
embodiments (e.g., multimode beams) can be approximated using an
approximation of the beam intensity distribution. For example, as
described more fully below, measurements of the beam intensity
distribution can be approximated by a Gaussian (1/e.sup.2
measurements) or by a "top hat" distribution and a selected
perimeter of the beam intensity distribution can be used to define
a bound of the area of the light beam. In certain embodiments, the
irradiance at the emission surface 22 is selected to provide the
desired irradiances at the subdermal target tissue. The irradiance
of the light beam is preferably controllably variable so that the
emitted light energy can be adjusted to provide a selected
irradiance at the subdermal tissue being treated. In certain
embodiments, the light beam emitted from the emission surface 22 is
continuous with a total radiant power in a range of about 4 Watts
to about 6 Watts. In certain embodiments, the radiant power of the
light beam is 5 Watts.+-.20% (CW). In certain embodiments, the peak
power for pulsed light is in a range of about 10 Watts to about 30
Watts (e.g., 20 Watts). In certain embodiments, the peak power for
pulsed light multiplied by the duty cycle of the pulsed light
yields an average radiant power in a range of about 4 Watts to
about 6 Watts (e.g., 5 Watts).
[0154] In certain embodiments, the time-averaged irradiance at the
subdermal target tissue (e.g., at a depth of approximately 2
centimeters below the dura) is at least about 0.01 mW/cm.sup.2 and
up to about 1 W/cm.sup.2 at the level of the tissue. In various
embodiments, the time-averaged subsurface irradiance at the target
tissue is at least about 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 15, 20,
30, 40, 50, 60, 70, 80, or 90 mW/cm.sup.2, depending on the desired
clinical performance. In certain embodiments, the time-averaged
subsurface irradiance at the target tissue is about 0.01
mW/cm.sup.2 to about 100 mW/cm.sup.2, about 0.01 mW/cm.sup.2 to
about 50 mW/cm.sup.2, about 2 mW/cm.sup.2 to about 20 mW/cm.sup.2,
or about 5 mW/cm.sup.2 to about 25 mW/cm.sup.2. In certain
embodiments, a duty cycle in a range between 1% and 80%, between
10% and 30%, or about 20% can be used with a peak irradiance at the
target tissue of 0.05 mW/cm.sup.2 to about 500 mW/cm.sup.2, about
0.05 mW/cm.sup.2 to about 250 mW/cm.sup.2, about 10 mW/cm.sup.2 to
about 100 mW/cm.sup.2, or about 25 mW/cm.sup.2 to about 125
mW/cm.sup.2.
[0155] In certain embodiments, the irradiance of the light beam is
selected to provide a predetermined irradiance at the subdermal
target tissue (e.g., at a depth of approximately 2 centimeters from
the dura). The selection of the appropriate irradiance of the light
beam emitted from the emission surface to use to achieve a desired
subdermal irradiance preferably includes consideration of
scattering by intervening tissue. Further information regarding the
scattering of light by tissue is provided by U.S. Pat. No.
7,303,578, which is incorporated in its entirety by reference
herein, and V. Tuchin in "Tissue Optics: Light Scattering Methods
and Instruments for Medical Diagnosis," SPIE Press (2000),
Bellingham, Wash., pp. 3-11, which is incorporated in its entirety
by reference herein.
[0156] Phototherapy for the treatment of neurologic conditions
(e.g., ischemic stroke, Alzheimer's Disease, Parkinson's Disease,
depression, or TBI) is based in part on the discovery that
irradiance or power density (i.e., power per unit area or number of
photons per unit area per unit time) and energy density (i.e.,
energy per unit area or number of photons per unit area) of the
light energy applied to tissue appear to be significant factors in
determining the relative efficacy of low level phototherapy. This
discovery is particularly applicable with respect to treating and
saving surviving but endangered neurons in a zone of danger
surrounding the primary injury. Certain embodiments described
herein are based at least in part on the finding that, given a
selected wavelength of light energy, it is the irradiance and/or
the energy density of the light delivered to tissue (as opposed to
the total power or total energy delivered to the tissue) that
appears to be important factors in determining the relative
efficacy of phototherapy.
[0157] Without being bound by theory or by a specific mechanism, it
is believed that light energy delivered within a certain range of
irradiances and energy densities provides the desired
biostimulative effect on the intracellular environment, such that
proper function is returned to previously nonfunctioning or poorly
functioning mitochondria in at-risk neurons. The biostimulative
effect may include interactions with chromophores within the target
tissue, which facilitate production of ATP and/or controls,
inhibits, prevents, minimizes, or reduces apoptosis of the injured
cells which have experienced decreased blood flow (e.g., due to the
stroke or TBI). Because strokes and TBI correspond to interruptions
of blood flow to portions of the brain, it is thought that any
effects of increasing blood flow by phototherapy are of less
importance in the efficacy of phototherapy for stroke or TBI
victims. Further information regarding the role of irradiance and
exposure time is described by Hans H.F.I. van Breugel and P. R. Dop
Bar in "Power Density and Exposure Time of He--Ne Laser Irradiation
Are More Important Than Total Energy Dose in Photo-Biomodulation of
Human Fibroblasts In Vitro," Lasers in Surgery and Medicine, Volume
12, pp. 528-537 (1992), which is incorporated in its entirety by
reference herein. In addition, the significance of the irradiance
used in phototherapy with regard to the devices and methods used in
phototherapy of brain tissue, are described more fully in U.S. Pat.
No. 7,303,578, U.S. Patent Appl. Publ. Nos. 2005/0107851 A1,
2007/0179570 A1, and 2007/0179571 A1, each of which is incorporated
in its entirety by reference herein. While these previous
discussions of irradiance were primarily in conjunction with
phototherapy of stroke, they apply as well to phototherapy of TBI.
For example, in certain embodiments, to obtain a desired average
power density at the brain for treating TBI, higher total power at
the scalp or skull can be used in conjunction with a larger spot
size at the scalp or skull. Thus, by increasing the spot size at
the scalp or skull, a desired average power density at the brain
can be achieved with lower power densities at the scalp or skull
which can reduce the possibility of overheating the scalp, skull,
or brain.
[0158] In certain embodiments, delivering the neuroprotective
amount of light energy includes selecting a surface irradiance of
the light energy at the scalp or skull corresponding to the
predetermined irradiance at the target area of the brain. As
described above, light propagating through tissue is scattered and
absorbed by the tissue. Calculations of the irradiance to be
applied to the scalp or skull so as to deliver a predetermined
irradiance to the selected target area of the brain preferably take
into account the attenuation of the light energy as it propagates
through the skin and other tissues, such as bone and brain tissue.
Factors known to affect the attenuation of light propagating to the
brain from the scalp or skull include, but are not limited to, skin
pigmentation, the presence, type, and color of hair over the area
to be treated, amount of fat tissue, the presence of bruised
tissue, skull thickness, patient's age and gender, and the location
of the target area of the brain, particularly the depth of the area
relative to the surface of the scalp or skull. (For a general
discussion of the absorption of light by melanins in the body, see,
e.g., "Optical Absorption Spectra of Melanins--a Comparison of
Theoretical and Experimental Results,"
accelrys.com/references/case-studies/melanins_partII.pdf) The
higher the level of skin pigmentation, the higher the irradiance
applied to the scalp to deliver a predetermined irradiance of light
energy to a subsurface site of the brain. The target area of the
patient's brain can be previously identified such as by using
standard medical imaging techniques.
[0159] The irradiance selected to be applied to the target area of
the patient's brain depends on a number of factors, including, but
not limited to, the wavelength of the applied light, the type of
CVA (ischemic or hemorrhagic), and the patient's clinical
condition, including the extent of the affected brain area. The
irradiance or power density of light energy to be delivered to the
target area of the patient's brain may also be adjusted to be
combined with any other therapeutic agent or agents, especially
pharmaceutical neuroprotective agents, to achieve the desired
biological effect. In such embodiments, the selected irradiance can
also depend on the additional therapeutic agent or agents
chosen.
Temporal Pulsewidth, Temporal Pulseshape, Duty Cycle, Repetition
Rate, and Irradiance Per Pulse
[0160] FIG. 21A schematically illustrates a generalized temporal
profile of a pulsed light beam in accordance with certain
embodiments described herein. The temporal profile comprises a
plurality of pulses (P.sub.1, P.sub.2, . . . , P.sub.i), each pulse
having a temporal pulsewidth during which the instantaneous
intensity or irradiance I(t) of the pulse is substantially
non-zero. For example, for the pulsed light beam of FIG. 21A, pulse
P.sub.1 has a temporal pulsewidth from time t=0 to time t=T.sub.1,
pulse P.sub.2 has a temporal pulsewidth from time t=T.sub.2 to time
t=T.sub.3, and pulse P.sub.1 has a temporal pulsewidth from time
t=T.sub.i to time t=T.sub.i+1. The temporal pulsewidth can also be
referred to as the "pulse ON time." The pulses are temporally
spaced from one another by periods of time during which the
intensity or irradiance of the beam is substantially zero. For
example, pulse P.sub.1 is spaced in time from pulse P.sub.2 by a
time t=T.sub.2-T.sub.1. The time between pulses can also be
referred to as the "pulse OFF time." In certain embodiments, the
pulse ON times of the pulses are substantially equal to one
another, while in certain other embodiments, the pulse ON times
differ from one another. In certain embodiments, the pulse OFF
times between the pulses are substantially equal to one another,
while in certain other embodiments, the pulse OFF times between the
pulses differ from one another. As used herein, the term "duty
cycle" has its broadest reasonable interpretation, including but
not limited to, the pulse ON time divided by the sum of the pulse
ON time and the pulse OFF time. For a pulsed light beam, the duty
cycle is less than one. The values of the duty cycle and the
temporal pulsewidth fully define the repetition rate of the pulsed
light beam.
[0161] Each of the pulses can have a temporal pulseshape which
describes the instantaneous intensity or irradiance of the pulse
I(t) as a function of time. For example, as shown in FIG. 21A, the
temporal pulseshapes of the pulsed light beam are irregular, and
are not the same among the various pulses. In certain embodiments,
the temporal pulseshapes of the pulsed light beam are substantially
the same among the various pulses. For example, as schematically
shown in FIG. 21B, the pulses can have a square temporal
pulseshape, with each pulse having a substantially constant
instantaneous irradiance over the pulse ON time. In certain
embodiments, the peak irradiances of the pulses differ from one
another (see, e.g., FIGS. 21A and 21B), while in certain other
embodiments, the peak irradiances of the pulses are substantially
equal to one another (see, e.g., FIGS. 21C and 21D). Various other
temporal pulseshapes (e.g., triangular, trapezoidal) are also
compatible with certain embodiments described herein. FIG. 21C
schematically illustrates a plurality of trapezoidal pulses in
which each pulse has a rise time (e.g., corresponding to the time
between an instantaneous irradiance of zero and a peak irradiance
of the pulse) and a fall time (e.g., corresponding to the time
between the peak irradiance of the pulse and an instantaneous
irradiance of zero). In certain embodiments, the rise time and the
fall time can be expressed relative to a specified fraction of the
peak irradiance of the pulse (e.g., time to rise/fall to 50% of the
peak irradiance of the pulse).
[0162] As used herein, the term "peak irradiance" of a pulse
P.sub.i has its broadest reasonable interpretation, including but
not limited to, the maximum value of the instantaneous irradiance
I(t) during the temporal pulsewidth of the pulse. In certain
embodiments, the instantaneous irradiance is changing during the
temporal pulsewidth of the pulse (see, e.g., FIGS. 21A and 21C),
while in certain other embodiments, the instantaneous irradiance is
substantially constant during the temporal pulsewidth of the pulse
(see, e.g., FIGS. 21B and 21D).
[0163] As used herein, the term "pulse irradiance" I.sub.P.sub.i of
a pulse P.sub.i has its broadest reasonable interpretation,
including but not limited to, the integral of the instantaneous
irradiance I(t) of the pulse P.sub.i over the temporal pulsewidth
of the pulse:
I P i = .intg. T i T i + 1 I ( t ) t / ( T i + 1 - T i ) .
##EQU00001##
As used herein, the term "total irradiance" I.sub.TOTAL has its
broadest reasonable interpretation, including but not limited to,
the sum of the pulse irradiances of the pulses:
I TOTAL = i = 0 N I P i . ##EQU00002##
As used herein, the term "time-averaged irradiance" I.sub.AVE has
its broadest reasonable interpretation, including but not limited
to, the integral of the instantaneous irradiance I(t) over a period
of time T large compared to the temporal pulsewidths of the
pulses:
I AVE = .intg. 0 T I ( t ) t / T . ##EQU00003##
The integral
.intg. 0 T I ( t ) t ##EQU00004##
provides the energy of the pulsed light beam.
[0164] For example, for a plurality of square pulses with different
pulse irradiances I.sub.P.sub.i and different temporal pulsewidths
.DELTA.T.sub.i, the time-averaged irradiance over a time T
equals
I AVE = 1 T i I P i .DELTA. T i . ##EQU00005##
For another example, for a plurality of square pulses with equal
pulse irradiances I.sub.P, with equal temporal pulsewidths, and
equal pulse OFF times (having a duty cycle D), the time-averaged
irradiance equals I.sub.AVE=I.sub.PD. For example, as shown in FIG.
21D, the time-averaged irradiance (shown as a dashed line) is less
than the pulse irradiance of the pulses.
[0165] The pulse irradiances and the duty cycle can be selected to
provide a predetermined time-averaged irradiance. In certain
embodiments in which the time-averaged irradiance is equal to the
irradiance of a continuous-wave (CW) light beam, the pulsed light
beam and the CW light beam have the same number of photons or flux
as one another. For example, a pulsed light beam with a pulse
irradiance of 5 mW/cm.sup.2 and a duty cycle of 20% provides the
same number of photons as a CW light beam having an irradiance of 1
mW/cm.sup.2. However, in contrast to a CW light beam, the
parameters of the pulsed light beam can be selected to deliver the
photons in a manner which achieve results which are not obtainable
using CW light beams.
[0166] For example, for hair removal, tattoo removal, or wrinkle
smoothing, pulsed light beams have previously been used to achieve
selective photothermolysis in which a selected portion of the skin
is exposed to sufficiently high temperatures to damage the hair
follicles (e.g., temperatures greater than 60 degrees Celsius), to
ablate the tattoo ink (e.g., temperatures much greater than 60
degrees Celsius), or to shrink the collagen molecules (e.g.,
temperatures between 60-70 degrees Celsius), respectively, while
keeping the other portions of skin at sufficiently low temperatures
to avoid unwanted damage or discomfort. The parameters of these
pulsed light beams are selected to achieve the desired elevated
temperature at the selected portion of the skin by absorption of
the light by the selected chromophore while allowing heat to
dissipate (characterized by a thermal relaxation time) during the
pulse OFF times to keep other areas of skin at lower temperatures.
As described by J. Lepselter et al., "Biological and clinical
aspects in laser hair removal," J. Dermatological Treatment, Vol.
15, pp. 72-83 (2004), the pulse ON time for hair removal is
selected to be between the thermal relaxation time for the
epidermis (about 3-10 milliseconds) and the thermal relaxation time
for the hair follicle (about 40-100 milliseconds). In this way, the
hair follicle can be heated to sufficiently high temperatures to
damage the follicle without causing excessive damage to the
surrounding skin.
[0167] In contrast to these treatments which are based on creating
thermal damage to at least a portion of the skin, certain
embodiments described herein utilize pulse parameters which do not
create thermal damage to at least a portion of the skin. In certain
embodiments, one or more of the temporal pulsewidth, temporal
pulseshape, duty cycle, repetition rate, and pulse irradiance of
the pulsed light beam are selected such that no portion of the skin
is heated to a temperature greater than 60 degrees Celsius, greater
than 55 degrees Celsius, greater than 50 degrees Celsius, or
greater than 45 degrees Celsius. In certain embodiments, one or
more of the temporal pulsewidth, temporal pulseshape, duty cycle,
repetition rate, and pulse irradiance of the pulsed light beam are
selected such that no portion of the skin is heated to a
temperature greater than 30 degrees Celsius above its baseline
temperature, greater than 20 degrees Celsius above its baseline
temperature, or greater than 10 degrees Celsius above its baseline
temperature. In certain embodiments, one or more of the temporal
pulsewidth, temporal pulseshape, duty cycle, repetition rate, and
pulse irradiance of the pulsed light beam are selected such that no
portion of the brain is heated to a temperature greater than 5
degrees Celsius above its baseline temperature, greater than 3
degrees Celsius above its baseline temperature, or greater than 1
degree Celsius above its baseline temperature. As used herein, the
term "baseline temperature" has its broadest reasonable
interpretation, including but not limited to, the temperature at
which the tissue would have if it were not irradiated by the light.
In contrast to previous low-light level therapies, the pulsed light
beam has an average radiant power in the range of about 1 Watt to
about 6 Watts or in a range of about 4 Watt to about 6 Watts.
[0168] In certain embodiments, the pulse parameters are selected to
achieve other effects beyond those which are achievable using CW
light beams. For example, while CW irradiation of brain cells in
vivo provides an efficacious treatment of stroke, the use of CW
irradiation for the treatment of TBI is more difficult, owing in
part to the excess blood within the region of the scalp, skull, or
cranium to be irradiated (e.g. due to intercranial bleeding). This
excess blood may be between the light source and the target brain
tissue to be irradiated, resulting in higher absorption of the
light applied to the scalp or skull before it can propagate to the
target tissue. This absorption can reduce the amount of light
reaching the target tissue and can unduly heat the intervening
tissue to an undesirable level.
[0169] In certain embodiments described herein, pulsed irradiation
may provide a more efficacious treatment. The pulsed irradiation
can provide higher peak irradiances for shorter times, thereby
providing more power to propagate to the target tissue while
allowing thermal relaxation of the intervening tissue and blood
between pulses to avoid unduly heating the intervening tissue. The
time scale for the thermal relaxation is typically in the range of
a few milliseconds. For example, the thermal relaxation time
constant (e.g. the time for tissue to cool from an elevated
temperature to one-half the elevated temperature) of human skin is
about 3-10 milliseconds, while the thermal relaxation time constant
of human hair follicles is about 40-100 milliseconds. Thus,
previous applications of pulsed light to the body for hair removal
have optimized temporal pulsewidths of greater than 40 milliseconds
with time between pulses of hundreds of milliseconds.
[0170] However, while pulsed light of this time scale
advantageously reduces the heating of intervening tissue and blood,
it does not provide an optimum amount of efficaciousness as
compared to other time scales. In certain embodiments described
herein, the patient's scalp or skull is irradiated with pulsed
light having parameters which are not optimized to reduce thermal
effects, but instead are optimized to stimulate, to excite, to
induce, or to otherwise support one or more intercellular or
intracellular biological processes which are involved in the
survival, regeneration, or restoration of performance or viability
of brain cells. Thus, in certain such embodiments, the selected
temporal profile can result in temperatures of the irradiated
tissue which are higher than those resulting from other temporal
profiles, but which are more efficacious than these other temporal
profiles. In certain embodiments, the pulsing parameters are
selected to utilize the kinetics of the biological processes rather
than optimizing the thermal relaxation of the tissue. In certain
embodiments, the pulsed light beam has a temporal profile (e.g.,
peak irradiance per pulse, a temporal pulse width, and a pulse duty
cycle) selected to modulate membrane potentials in order to
enhance, restore, or promote cell survival, cell function, or both
of the irradiated brain cells following the traumatic brain injury.
For example, in certain embodiments, the pulsed light has a
temporal profile which supports one or more intercellular or
intracellular biological processes involved in the survival or
regeneration of brain cells, but does not optimize the thermal
relaxation of the irradiated tissue. In certain embodiments, the
brain cells survive longer after the irradiation as compared to
their survival if the irradiation did not occur. For example, the
light of certain embodiments can have a protective effect on the
brain cells, or can cause a regeneration process in the brain
cells.
[0171] In certain embodiments, the temporal profile (e.g., peak
irradiance, temporal pulse width, and duty cycle) are selected to
utilize the kinetics of the biological processes while maintaining
the irradiated portion of the scalp or skull at or below a
predetermined temperature. This predetermined temperature is higher
than the optimized temperature which could be achieved for other
temporal profiles (e.g., other values of the peak irradiance,
temporal pulse width, and duty cycle) which are optimized to
minimize the temperature increase of surrounding tissue due to the
irradiation. For example, a temporal profile having a peak
irradiance of 10 W/cm.sup.2 and a duty cycle of 20% has a
time-averaged irradiance of 2 W/cm.sup.2. Such a pulsed light beam
provides the same number of photons to the irradiated surface as
does a continuous-wave (CW) light beam with an irradiance of 2
W/cm.sup.2. However, because of the "dark time" between pulses, the
pulsed light beam can result in a lower temperature increase than
does the CW light beam. To minimize the temperature increase of the
irradiated portion of the scalp or skull, the temporal pulse width
and the duty cycle can be selected to allow a significant portion
of the heat generated per pulse to dissipate before the next pulse
reaches the irradiated portion. In certain embodiments described
herein, rather than optimizing the beam temporal parameters to
minimize the temperature increase, the temporal parameters are
selected to effectively correspond to or to be sufficiently close
to the timing of the biomolecular processes involved in the
absorption of the photons to provide an increased efficacy. Rather
than having a temporal pulse width on the order of hundreds of
microseconds, certain embodiments described herein utilize a
temporal pulse width which does not optimize the thermal relaxation
of the irradiated tissue (e.g., milliseconds, tens of milliseconds,
hundreds of milliseconds). Since these pulse widths are
significantly longer than the thermal relaxation time scale, the
resulting temperature increases are larger than those of smaller
pulse widths, but still less than that of CW light beams due to the
heat dissipation the time between the pulses.
[0172] A number of studies have investigated the effects of in
vitro irradiation of cells using pulsed light on various aspects of
the cells. A study of the action mechanisms of incoherent pulsed
radiation at a wavelength of 820 nanometers (pulse repetition
frequency of 10 Hz, pulse width of 20 milliseconds, dark period
between pulses of 80 milliseconds, and duty factor (pulse duration
to pulse period ratio) of 20%) on in vitro cellular adhesion has
found that pulsed infrared radiation at 820 nanometers increases
the cell-matrix attachment. (T. I. Karu et al., "Cell Attachment to
Extracellular Matrices is Modulated by Pulsed Radiation at 820 nm
and Chemicals that Modify the Activity of Enzymes in the Plasma
Membrane," Lasers in Surgery and Medicine, Vol. 29, pp. 274-281
(2001) which is incorporated in its entirety by reference herein.)
It was hypothesized in this study that the modulation of the
monovalent ion fluxes through the plasma membrane, and not the
release of arachidonic acid, is involved in the cellular signaling
pathways activated by irradiation at 820 nanometers. A study of
light-induced changes to the membrane conductance of ventral
photoreceptor cells found behavior which was dependent on the pulse
parameters, indicative of two light-induced membrane processes. (J.
E. Lisman et al., "Two Light-Induced Processes in the Photoreceptor
Cells of Limulus Ventral Eye," J. Gen. Physiology, Vol. 58, pp.
544-561 (1971), which is incorporated in its entirety by reference
herein.) Studies of laser-activated electron injection into
oxidized cytochrome c oxidase observed kinetics which establish the
reaction sequence of the proton pump mechanism and some of its
thermodynamic properties have time constants on the order of a few
milliseconds. (I. Belevich et al., "Exploring the proton pump
mechanism of cytochrome c oxidase in real time," Proc. Nat'l Acad.
Sci., Vol. 104, pp. 2685-2690 (2007); I. Belevich et al.,
"Proton-coupled electron transfer drives the proton pump of
cytochrome coxidase," Nature, Vol. 440, pp. 829-832 (2006), both of
which are incorporated in its entirety by reference herein.) An in
vivo study of neural activation based on pulsed infrared light
proposed a photo-thermal effect from transient tissue temperature
changes resulting in direct or indirect activation of transmembrane
ion channels causing propagation of the action potential. (J. Wells
et al., "Biophysical mechanisms responsible for pulsed low-level
laser excitation of neural tissue," Proc. SPIE, Vol. 6084, pp.
60840X (2006), which is incorporated in its entirety by reference
herein.)
[0173] In certain embodiments, the temporal profile of the pulsed
light beam comprises a peak irradiance, a temporal pulse width, a
temporal pulse shape, a duty cycle, and a pulse repetition rate or
frequency. In certain embodiments in which the pulsed light beam is
transmitted through a region of the scalp or skull containing an
excess amount of hemorrhagic blood due to the at least one physical
trauma (e.g. due to intercranial bleeding), at least one of the
peak irradiance, temporal pulse width, temporal pulse shape, duty
cycle, and pulse repetition rate or frequency is selected to
provide a time-averaged irradiance (averaged over a time period
including a plurality of pulses) at the emission surface 22 of the
output optical assembly 20 between about 10 mW/cm.sup.2 to about 10
W/cm.sup.2, between about 100 mW/cm.sup.2 to about 1000
mW/cm.sup.2, between about 500 mW/cm.sup.2 to about 1 W/cm.sup.2,
or between about 650 mW/cm.sup.2 to about 750 mW/cm.sup.2 across
the cross-sectional area of the light beam. In certain such
embodiments, the time-averaged irradiance at the brain cells being
treated (e.g., at a depth of approximately 2 centimeters below the
dura) is greater than 0.01 mW/cm.sup.2.
[0174] In certain embodiments, the peak irradiance per pulse across
the cross-sectional area of the light beam at the emission surface
22 of the output optical assembly 20 is in a range between about 10
mW/cm.sup.2 to about 10 W/cm.sup.2, between about 100 mW/cm.sup.2
to about 1000 mW/cm.sup.2, between about 500 mW/cm.sup.2 to about 1
W/cm.sup.2, between about 650 mW/cm.sup.2 to about 750 mW/cm.sup.2,
between about 20 mW/cm.sup.2 to about 20 W/cm.sup.2, between about
200 mW/cm.sup.2 to about 2000 mW/cm.sup.2, between about 1
W/cm.sup.2 to about 2 W/cm.sup.2, between about 1300 mW/cm.sup.2 to
about 1500 mW/cm.sup.2, between about 1 W/cm.sup.2 to about 1000
W/cm.sup.2, between about 10 W/cm.sup.2 to about 100 W/cm.sup.2,
between about 50 W/cm.sup.2 to about 100 W/cm.sup.2, or between
about 65 W/cm.sup.2 to about 75 W/cm.sup.2. In certain embodiments,
the temporal pulse shape is generally rectangular, generally
triangular, or any other shape. In certain embodiments, the pulses
have a rise time (e.g., from 10% of the peak irradiance to 90% of
the peak irradiance) less than 1% of the pulse ON time, or a fall
time (e.g., from 90% of the peak irradiance to 10% of the peak
irradiance) less than 1% of the pulse ON time.
[0175] In certain embodiments, the pulses have a temporal
pulsewidth (e.g., pulse ON time) in a range between about 0.001
millisecond and about 150 seconds, between about 0.01 millisecond
and about 10 seconds, between about 0.1 millisecond and about 1
second, between about 0.5 millisecond and about 100 milliseconds,
between about 2 milliseconds and about 20 milliseconds, or between
about 1 millisecond and about 10 milliseconds. In certain
embodiments, the pulse width is about 0.5, 1, 2, 4, 6, 8, 10, 15,
20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 140, 160, 180, 200, 220,
240, 260, 280, or 300 milliseconds. In certain embodiments, the
temporal pulsewidth is in a range between about 0.1 millisecond and
150 seconds.
[0176] In certain embodiments, the time between pulses (e.g., pulse
OFF time) is in a range between about 0.01 millisecond and about
150 seconds, between about 0.1 millisecond and about 100
millisecond, between about 4 milliseconds and about 1 second,
between about 8 milliseconds and about 500 milliseconds, between
about 8 milliseconds and about 80 milliseconds, or between about 10
milliseconds and about 200 milliseconds. In certain embodiments,
the time between pulses is about 4, 8, 10, 20, 50, 100, 200, 500,
700, or 1000 milliseconds.
[0177] In certain embodiments, the pulse duty cycle is in a range
between about 1% and about 80% or in a range between about 10% and
about 30%. In certain embodiments, the pulse duty cycle is about
10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%.
Beam Size and Beam Profile
[0178] In certain embodiments, the light beam emitted from the
output optical assembly 20 has a nominal diameter in a range of
about 10 millimeters to about 40 millimeters, in a range of about
20 millimeters to about 35 millimeters, or equal to about 30
millimeters. In certain embodiments, the cross-sectional area is
generally circular with a radius in a range of about 1 centimeter
to about 2 centimeters. In certain embodiments, the light beam
emitted from the emission surface 22 has a cross-sectional area
greater than about 2 cm.sup.2 or in a range of about 2 cm.sup.2 to
about 20 cm.sup.2 at the emission surface 22 of the optical element
23. In certain embodiments, the output optical element 23 has an
aperture diameter of less than 33 millimeters.
[0179] As used herein, the beam diameter is defined to be the
largest chord of the perimeter of the area of the scalp or skull
irradiated by the light beam at an intensity of at least 1/e.sup.2
of the maximum intensity of the light beam. The perimeter of the
light beam used to determine the diameter of the beam is defined in
certain embodiments to be those points at which the intensity of
the light beam is 1/e.sup.2 of the maximum intensity of the light
beam. The maximum-useful diameter of certain embodiments is limited
by the size of the patient's head and by the heating of the
patient's head by the irradiation. The minimum-useful diameter of
certain embodiments is limited by heating and by the total number
of treatment sites that could be practically implemented. For
example, to cover the patient's skull with a beam having a small
beam diameter would correspondingly use a large number of treatment
sites. In certain embodiments, the time of irradiation per
treatment site can be adjusted accordingly to achieve a desired
exposure dose.
[0180] Specifying the total flux inside a circular aperture with a
specified radius centered on the exit aperture ("encircled energy")
is a method of specifying the power (irradiance) distribution over
the light beam emitted from the emission surface 22. The "encircled
energy" can be used to ensure that the light beam is not too
concentrated, too large, or too small. In certain embodiments, the
light beam emitted from the emission surface has a total radiant
power, and the light beam has a total flux inside a 20-millimeter
diameter cross-sectional circle centered on the light beam at the
emission surface 22 which is no more than 75% of the total radiant
power. In certain such embodiments, the light beam has a total flux
inside a 26-millimeter diameter cross-sectional circle centered on
the light beam at the emission surface 22 which is no less than 50%
of the total radiant power.
[0181] In certain embodiments, the beam intensity profile has a
semi-Gaussian profile, while in certain other embodiments, the beam
intensity profile has a "top hat" profile. In certain embodiments,
the light beam is substantially without high flux regions or "hot
spots" in the beam intensity profile in which the local flux,
averaged over a 3 millimeter by 3 millimeter area, is more than 10%
larger than the average flux. Certain embodiments of the apparatus
10 advantageously generate a light beam substantially without hot
spots, thereby avoiding large temperature gradients at the
patient's skin which would otherwise cause discomfort to the
patient.
Divergence
[0182] In certain embodiments, the beam divergence emitted from the
emission surface 22 is significantly less than the scattering angle
of light inside the body tissue being irradiated, which is
typically several degrees. In certain embodiments, the light beam
has a divergence angle greater than zero and less than 35
degrees.
[0183] As the distance between a light source and an observer
increases, the diameter of the source becomes less relevant to
considerations of the beam divergence. For example, an end of the
optical fiber 40 providing the light has a diameter of about 1
millimeter. At a close distance, observing from a specific
location, light rays from the edges of the optical fiber end can
arrive at the observation point with significantly different
angles. However, as the observation point moves away from the light
source, this angular discrepancy is reduced and the source appears
more like a point source.
[0184] In certain embodiments, with the output optical assembly 20
mounted onto the apparatus 10, the optical distance between the
emission surface 22 and the end of the optical fiber 40 is about
82.7 millimeters. The beam divergence dictated by the numerical
aperture of the optical fiber 40 and the exit aperture of the
optical element 23 is about 23 degrees. In certain embodiments,
with the output optical assembly 20 not mounted onto the apparatus
10, the optical distance between the window 70 and the end of the
optical fiber is about 57.5 millimeters, and the beam divergence
dictated by the numerical aperture of the optical fiber 40 and the
exit aperture of the window 70 is about 16 degrees. With a source
diameter of about 1 millimeter, the angular ambiguity in the beam
divergence is about .+-.0.35 degree. Thus, the angular ambiguity is
much less than the beam divergence angle regardless of whether the
output optical assembly 20 is mounted or not onto the apparatus 10,
so the optical fiber 40 can be treated as a point source. In
certain such embodiments, the beam divergence or radiant intensity
(e.g., measured in Watts/steradian) can be calculated directly from
the beam profile or from the irradiance.
Treatment Time
[0185] In certain embodiments, the treatment per treatment site
proceeds continuously for a period of about 10 seconds to about 2
hours, for a period of about 1 to about 10 minutes, or for a period
of about 1 to 5 minutes. For example, the treatment time per
treatment site in certain embodiments is about two minutes. In
other embodiments, the light energy is delivered for at least one
treatment period of at least about five minutes, or for at least
one treatment period of at least ten minutes. The minimum treatment
time of certain embodiments is limited by the biological response
time (which is on the order of microseconds). The maximum treatment
time of certain embodiments is limited by heating and by practical
treatment times (e.g., completing treatment within about 24 hours
of stroke onset). The light energy can be pulsed during the
treatment period or the light energy can be continuously applied
during the treatment period. If the light is pulsed, the pulses can
be 2 milliseconds long and occur at a frequency of 100 Hz, although
longer pulselengths and lower frequencies can be used, or at least
about 10 nanosecond long and occur at a frequency of up to about
100 kHz.
[0186] In certain embodiments, the treatment may be terminated
after one treatment period, while in other embodiments, the
treatment may be repeated for at least two treatment periods. The
time between subsequent treatment periods can be at least about
five minutes, at least two in a 24-hour period, at least about 1 to
2 days, or at least about one week. The length of treatment time
and frequency of treatment periods can depend on several factors,
including the functional recovery of the patient and the results of
imaging analysis of the injury (e.g., infarct). In certain
embodiments, one or more treatment parameters can be adjusted in
response to a feedback signal from a device (e.g., magnetic
resonance imaging) monitoring the patient.
Cooling Parameters
[0187] In certain embodiments, the apparatus 10 comprises an output
optical element 23 in optical communication with a source of light.
The output optical element 23 comprises an emission surface 22
configured to emit a light beam in accordance with the light
parameters disclosed above. In certain embodiments the apparatus 10
further comprises a thermally conductive portion configured to be
placed in thermal communication with the irradiated portion of the
patient's scalp or skull and to remove heat from the irradiated
portion of the patient's scalp or skull. In certain embodiments,
the thermally conductive portion comprises the output optical
element 23. The thermally conductive portion of certain embodiments
is releasably coupled to the output optical element 23.
[0188] In certain embodiments, the thermally conductive portion
removes heat from the irradiated portion of the patient's scalp or
skull. This cooling of the scalp or skull can to improve the
comfort of the patient, by controlling, inhibiting, preventing,
minimizing, or reducing temperature increases at the scalp or skull
due to the irradiation. Thus, by virtue of the cooling of the
portion of the patient's scalp or skull being irradiated, the
temperature of the irradiated portion of the patient's scalp or
skull is lower than it would otherwise be if the irradiated portion
of the scalp or skull were not cooled. For example, by cooling the
irradiated portion of the patient's scalp or skull, the temperature
of the irradiated portion of the patient's scalp or skull can be
higher than the temperature of the portion of the patient's scalp
or skull if it were not irradiated, but lower than the temperature
of the portion of the patient's scalp or skull if it were
irradiated but not cooled. In addition, this cooling of the scalp
or skull can be to perform double-blind studies of the efficacy of
the phototherapy treatment by masking any heating of the scalp or
skull due to the irradiation. (See, e.g., B. Catanzaro et al.,
"Managing Tissue Heating in Laser Therapy to Enable Double-Blind
Clinical Study," Mechanisms for Low-Light Therapy, Proc. of the
SPIE, Vol. 6140, pp. 199-208 (2006).)
[0189] In certain embodiments, heat is removed from the irradiated
portion of the patient's scalp or skull by the thermally conductive
portion at a rate in a range of about 0.1 Watt to about 5 Watts or
in a range of about 1 Watt to about 3 Watts. In certain
embodiments, the thermally conductive portion is configured to
maintain the temperature of the irradiated portion of the patient's
scalp or skull to be less than 42 degrees Celsius. The thermally
conductive portion of certain embodiments is in thermal
communication with the emission surface 22 and is configured to
maintain the temperature of the emission surface to be in a range
of 18 degrees Celsius to 25 degrees Celsius under a heat load of 2
Watts. For a general description of cooling of the scalp, see,
e.g., F. E. M. Janssen et al., "Modeling of temperature and
perfusion during scalp cooling," Phys. Med. Biol., Vol. 50, pp.
4065-4073 (2005). In certain embodiments in which pulsed light is
used, the rate of heat removal can be less, or cooling may not be
utilized for certain ranges of pulsed dosimetries and timing.
Pressure Parameters
[0190] In certain embodiments, the apparatus 10 is configured to
have the thermally conductive portion move relative to a second
portion of the apparatus 10 upon a pressure being applied to the
thermally conductive portion above a predetermined threshold
pressure in a direction of movement of the thermally conductive
portion relative to the second portion of the apparatus 10. The
predetermined threshold pressure is sufficient to have the
thermally conductive portion in thermal communication with the
portion of the patient's scalp or skull. In certain such
embodiments, the apparatus 10 comprises a sensor configured to be
responsive to the movement of the thermally conductive portion
relative to the second portion by generating a signal (e.g.,
binary, analog, or digital) indicative of the movement.
[0191] In certain such embodiments, the sensor 130 in conjunction
with the trigger force spring 140 and the trigger force adjustment
mechanism 142 provides a mechanism for detecting whether the
apparatus 10 is being applied to the patient's scalp or skull with
a pressure above the predetermined threshold pressure. In certain
such embodiments, the sensor 130 detects movement between the first
portion of the apparatus 10 and the second portion of the apparatus
10 upon placing the emission surface 22 in thermal communication
with the patient's scalp or skull with sufficient pressure to
overcome the restoring force of the trigger force spring 140. Upon
applying the threshold pressure to the emission surface 22 move the
first and second portions relative to one another, the sensor 130
detects the movement and generates a corresponding signal. In
certain embodiments, the apparatus 10 further comprises a
controller operatively coupled to the light source and to the
sensor 130. The controller is configured to receive the signal from
the sensor 130 and to turn on the light source in response to the
signal being indicative of the pressure being above the
predetermined threshold pressure.
[0192] In certain embodiments, the threshold pressure is set to be
a pressure which results in blanching of the portion of the
patient's scalp to be irradiated. In certain embodiments, the
threshold pressure is 0.1 pound per square inch, while in certain
other embodiments, the threshold pressure is one pound per square
inch or about two pounds per square inch.
[0193] In certain embodiments in which pulsed light is used, the
amount of blanching can be less, or blanching may not be utilized
for certain ranges of pulsed dosimetries and timing. For example,
in certain embodiments, the patient may have a heightened
sensitivity to pressure applied to the scalp or skull (e.g., a TBI
patient). Thus, in certain embodiments, the apparatus 10 does not
apply sufficient pressure to the scalp of the patient (e.g.,
applies no pressure to the patient's scalp) to blanch the
irradiated portion of the scalp during the irradiation. In certain
other embodiments in which some amount of blanching of the
irradiated portion of the scalp is desired, the maximum pressure
used to blanch the irradiated portion of the scalp is limited by
patient comfort levels and tissue damage levels. For example, the
cranium or skull of a TBI patient may have cracks or breaks such
that the brain would be adversely affected if pressure were applied
to the scalp. The amount of pressure used, if any, is determined at
least in part, on the amount of pressure that the patient can
withstand without additional damage being done by the application
of pressure.
Irradiating Multiple Portions of the Scalp or Skull
[0194] FIGS. 22A-22C schematically illustrate an embodiment in
which the apparatus 10 is placed in thermal communication
sequentially with a plurality of treatment sites corresponding to
portions of the patient's scalp. In certain such embodiments, the
light emitted from the emission surface 22 propagates through the
scalp to the brain and disperses in a direction generally parallel
to the scalp, as shown in FIG. 22A. In certain embodiments in which
the patient is suffering from a TBI, one or more of the treatment
sites has a portion of the skull is exposed and at least a portion
of the light is applied to the exposed portion of the skull without
propagating through scalp tissue. In certain embodiments, the
treatment sites of the patient's scalp do not overlap one another.
The treatment sites (e.g., twenty treatment sites) are preferably
spaced sufficiently far apart from one another such that the light
emitted from the emission surface 22 to irradiate a treatment site
of the patient's scalp is transmitted through intervening tissue to
irradiate an area of the patient's brain which overlaps one or more
areas of the target tissue of the patient's brain irradiated by the
light emitted from the emission surface 22 when a neighboring
treatment site of the patient's scalp is irradiated. FIG. 22B
schematically illustrates this overlap as the overlap of circular
spots 160 across the target tissue at a reference depth at or below
the surface of the brain. FIG. 22C schematically illustrates this
overlap as a graph of the irradiance at the reference depth of the
brain along the line L-L of FIGS. 22A and 22B. Summing the
irradiances from the neighboring treatment sites (shown as a dashed
line in FIG. 22C) serves to provide a more uniform light
distribution at the target tissue to be treated. In such
embodiments, the summed irradiance is preferably less than a damage
threshold of the brain and above an efficacy threshold. In certain
embodiments, portions of the brain irradiated by irradiating the
treatment sites at the scalp do not overlap one another. In certain
such embodiments, the treatment sites at the scalp are positioned
so as to irradiate as much of the cortex as possible.
Example Wearable Apparatus
[0195] FIG. 23 schematically illustrates an example apparatus 500
which is wearable by a patient for treating the patient's brain.
The apparatus 500 comprises a body 510 and a plurality of
indicators 520. The body 510 is adapted to be worn over at least a
portion of the patient's scalp when the apparatus 500 is worn by
the patient. The plurality of indicators 520 correspond to a
plurality of treatment site location at the patient's scalp where
light is to be applied to irradiate at least a portion of the
patient's brain. At least one indicator 520 comprises a portion of
the body which is substantially transmissive (e.g., substantially
transparent or substantially translucent) to light emitted from the
emission surface 22 to irradiate at least a portion of the
patient's brain.
[0196] In certain embodiments, at least one of the indicators 520
denotes a position within an area of the patient's scalp
corresponding to a treatment site location. In certain such
embodiments, the position is the center of the area of the
patient's scalp. The adjacent treatment sites of certain
embodiments have areas which do not overlap one another or have
perimeters which are spaced from one another. In certain such
embodiments, the perimeters are spaced from one another by at least
10 millimeters or at least 25 millimeters.
[0197] In certain embodiments, each indicator 520 comprises an
opening or aperture through the body 510 at which the beam delivery
apparatus 10 can be placed to irradiate the portion of the
patient's scalp exposed by the hole or aperture. In certain
embodiments, the aperture has a substantially circular perimeter
and a diameter in a range between 20 millimeters and 50 millimeters
or in a range between 25 millimeters and 35 millimeters. In certain
embodiments, the aperture has a substantially elliptical perimeter
with a minor axis greater than 20 millimeters and a major axis less
than 50 millimeters. Other shapes of the aperture are also
compatible with certain embodiments described herein.
[0198] In certain embodiments, the plurality of indicators 520
comprises at least about 10 indicators 520 distributed across the
patient's scalp, while in certain other embodiments, the plurality
of indicators 520 comprises 20 indicators 520. In certain other
embodiments, the plurality of indicators 520 comprises between 15
and 25 indicators 520. In certain embodiments, the optically
transmissive portion of each indicator 520 has an area of at least
1 cm.sup.2, in a range between 1 cm.sup.2 and 20 cm.sup.2, or in a
range between 5 cm.sup.2 and 10 cm.sup.2.
[0199] In certain embodiments, the body 510 comprises a hood, while
in other embodiments, the body 510 comprises a cap or has another
configuration which is wearable on the patient's head and serves as
a support for orienting the indicators 520 on the patient's head.
In certain embodiments, the body 510 comprises a stretchable or
pliant material which generally conforms to the patient's scalp. In
certain embodiments, the body 510 comprises nylon-backed
polychloroprene or Tyvek.RTM.. In certain embodiments, the body 510
is available in different sizes (e.g. small, medium, large) to
accommodate different sizes of heads. In certain embodiments, the
body 510 is disposable after a single use to advantageously avoid
spreading infection or disease between subsequent patients.
[0200] The indicators 520 of certain embodiments are configured to
guide an operator to irradiate the patient's scalp at the
corresponding treatment site locations sequentially one at a time
in a predetermined order. In certain embodiments, the wearable
apparatus 500 further comprises a plurality of labels 522 with each
label in proximity to a corresponding indicator. The labels 522
advantageously provide one or more numbers, letters, or symbols
(e.g., bar codes) to each of the indicators 520 to distinguish the
various indicators 520 from one another. In certain such
embodiments, the labels 522 are mechanically coupled to the
corresponding indicators so as to be visible to users of the beam
delivery apparatus 10.
[0201] FIGS. 24A and 24B schematically illustrate the left-side and
right-side of an example apparatus 500, respectively, with labels
522 substantially covering the indicators 520 corresponding to the
treatment sites. In certain embodiments, the labels 522 are
advantageously used to keep track of which treatment sites have
been irradiated and which treatment sites are yet to be irradiated.
In certain such embodiments, at least a portion of each label 522
comprises a portion of the body (e.g., a pull-off tab or flap)
which is configured to be removed from the apparatus 500 when the
treatment site corresponding to the indicator 520 has been
irradiated. In certain embodiments, the labels 522 comprise
removable portions of the body 510 which cover the corresponding
indicator 520. In certain such embodiments, prior to irradiating
the treatment site location corresponding to the indicator 520, the
corresponding label 522 can be removed to allow access to the
underlying portion of the patient's scalp.
[0202] In certain embodiments, the label 522 has a code sequence
which the operator enters into the controller prior to irradiation
so as to inform the controller of which treatment site is next to
be irradiated. In certain other embodiments, each label 522
comprises a bar code or a radio-frequency identification device
(RFID) which is readable by a sensor electrically coupled to the
controller. The controller of such embodiments keeps track of which
treatment sites have been irradiated, and in certain such
embodiments, the controller only actuates the light source when the
beam delivery apparatus 10 is in optical and thermal communication
with the proper treatment site of the patient's scalp.
[0203] FIG. 24C schematically illustrates an example labeling
configuration from above a flattened view of the apparatus 500 of
FIGS. 24A and 24B. The labeling convention of FIG. 24C is
compatible with irradiation of both halves or hemispheres of the
patient's brain. Other labeling conventions are also compatible
with embodiments described herein.
[0204] In certain embodiments, the labels 522 are advantageously
used to guide an operator to irradiate the patient's brain at the
various treatment sites sequentially at each of the treatment sites
one at a time through the indicators 520 in a predetermined order
by optically and thermally coupling the beam delivery apparatus 10
to sequential treatment sites corresponding to the indicators 520.
For example, for the labeling configuration of FIG. 24C, the
operator can first irradiate treatment site "1," followed by
treatment sites "2," "3," "4," etc. to sequentially irradiate each
of the twenty treatment sites one at a time. In certain such
embodiments, the predetermined order of the treatment sites is
selected to advantageously reduce temperature increases which would
result from sequentially irradiating treatment sites in proximity
to one another.
[0205] In certain embodiments, the predetermined order comprises
irradiation of a first treatment site location on a first side of
the patient's scalp (e.g., site "2" of FIG. 24C), then irradiation
of a second treatment site location on a second side of the
patient's scalp (e.g., site "3" of FIG. 24C), then irradiation of a
third treatment site location on the first side of the patient's
scalp (e.g., site "4" of FIG. 24C). In certain such embodiments,
the predetermined order further comprises irradiation of a fourth
treatment site location on the second side of the patient's scalp
after irradiation of the third treatment site location. In certain
embodiments, two sequentially irradiated treatment site locations
are separated from one another by at least 25 millimeters.
[0206] For example, in certain embodiments, the predetermined order
comprises at least a portion of the following sequence of treatment
sites:
[0207] 1. Right anterior frontal
[0208] 2. Left lateral frontal
[0209] 3. Right anteroinferior parietal
[0210] 4. Left posterior mid-parietal
[0211] 5. Right superior parietal
[0212] 6. Right lateral frontal
[0213] 7. Left anterior frontal
[0214] 8. Left posterior superior parietal
[0215] 9. Left posteroinferior parietal
[0216] 10. Right posteroinferior parietal
[0217] 11. Right posterior superior parietal
[0218] 12. Right anterior mid-parietal
[0219] 13. Left anteroinferior parietal
[0220] 14. Left anterosuperior frontal
[0221] 15. Left superior occipital
[0222] 16. Left anterior mid-parietal
[0223] 17. Right posterior mid-parietal
[0224] 18. Right anterosuperior frontal
[0225] 19. Right superior occipital
[0226] 20. Left superior parietal
[0227] For example, the predetermined order of certain embodiments
comprises two, three, four, or more of these treatment sites in the
relative order listed above. The sequence of treatment sites of
certain embodiments comprises two, three, four, or more of these
treatment sites in a relative order which is the reverse of the
sequence listed above. While certain embodiments utilize at least a
portion the relative order listed above without irradiation at an
additional treatment site between two sequentially listed treatment
sites, certain other embodiments utilize at least a portion of the
relative order listed above with one or more additional treatment
sites between two of the sequentially listed treatment sites. In
certain embodiments, the exact anatomic locations of each treatment
site may be adjusted from those listed above to account for
variations among the sizes of the heads of the patients (e.g., very
large or very small). Thus, in certain embodiments, there is some
variability regarding the locations of the treatment sites for any
given individual.
[0228] In certain embodiments, the apparatus 500 serves as a
template for marking the patient's scalp to indicate the treatment
site locations. The apertures of the apparatus 500 can be used to
guide a user place marks on the patient's scalp, and the apparatus
500 can then be removed from the patient's scalp before the beam
delivery apparatus 10 is applied to the scalp for irradiating the
patient's brain. The marks remain on the patient's scalp to guide
the operator while the patient's brain is irradiated.
Methods of Light Delivery
[0229] FIGS. 25-28 are flow diagrams of example methods for
irradiating a surface with light. As described more fully below,
the methods are described by referring to the beam delivery
apparatus 10 and components thereof, as described herein. Other
configurations of a beam delivery apparatus are also compatible
with the methods in accordance with embodiments described
herein.
[0230] The method 610 of FIG. 25 comprises providing a beam
delivery apparatus 10 in an operational block 612. The beam
delivery apparatus 10 comprises a first portion and a second
portion mechanically coupled to the first portion and in optical
communication with the first portion, wherein the first portion and
the second portion are configured to move relative to one another,
as described more fully above. The method 610 further comprises
placing the second portion in thermal communication with the
surface in an operational block 614 (e.g., releasably operatively
coupling the second portion to the surface). The method 610 further
comprises irradiating the surface such that the light from the
first portion propagates through the second portion in an
operational block 616. The method 610 further comprises moving the
first portion and the second portion relative to one another in
response to the second portion being placed in thermal
communication with the surface in an operational block 618.
[0231] The method 620 of FIG. 26 comprises providing an optical
element 23 in an operational block 622. The optical element 23
comprises a substantially optically transmissive and substantially
thermally conductive material, and the optical element 23 has a
first surface 22 and a second surface 24, as described more fully
above. The method 620 further comprises placing the first surface
22 in thermal communication with the irradiated surface in an
operational block 624 (e.g., releasably operatively coupling the
first surface 22 to the irradiated surface). The method 620 further
comprises propagating the light along a first optical path 32
through the second surface 24 and through the first surface 22 to
the irradiated surface in an operational block 626. The method 620
further comprises detecting radiation propagating along a second
optical path 82 from at least a portion of the second surface 24,
wherein the first optical path 32 and the second optical path 82
have a non-zero angle therebetween in an operational block 628. In
certain embodiments, the first surface 22 and the second surface 24
face in generally opposite directions, and the first surface 22 is
not along the second optical path 82.
[0232] The method 630 of FIG. 27 comprises providing a
thermoelectric assembly 90 in an operational block 632. The
thermoelectric assembly 90 comprises a first surface 93 and a
second surface 94, and the thermoelectric assembly 90 generally
surrounds a first region 97, as described more fully above. The
method 630 further comprises providing an output optical assembly
20 in an operational block 633. The method 630 further comprises
releasably mechanically coupling the first surface 93 of the
thermoelectric assembly 90 to the output optical assembly 20 so
that the first surface 93 is in thermal communication with the
output optical assembly 20 in an operational block 634. The method
630 further comprises cooling the first surface 93 and heating the
second surface 94 in an operational block 636. The method 630
further comprises propagating light through the first region 97 to
impinge the irradiated surface in an operational block 638. In
certain embodiments, the first surface 22 and the second surface 24
face in generally opposite directions, and the first surface 22 is
not along the second optical path 82.
[0233] In certain embodiments, the output optical assembly 20
comprises an optical element 23 and a thermally conductive portion
25 generally surrounding a second region 28. The thermally
conductive portion 25 is in thermal communication with the optical
element 23. In certain such embodiments, releasably mechanically
coupling the first surface 93 to the output optical assembly 20
comprises releasably mechanically coupling the first surface 93 to
the thermally conductive portion 25. In certain such embodiments,
the method 630 further comprises placing the optical element 23 in
thermal communication with the irradiated surface and propagating
the light comprises transmitting the light through the first region
97, the second region 28, and the optical element 23 to impinge the
irradiated surface. In certain embodiments, the method 630 further
comprises providing a heat sink 100 in thermal communication with
the second surface 94 of the thermoelectric assembly 90. The heat
sink 100 generally surrounds a third region 107, and propagating
the light comprises transmitting the light through the third region
107, the first region 97, the second region 28, and the optical
element 23.
[0234] The method 640 of FIG. 28 comprises emitting a light beam
from an emission surface 22 of an optical element 23 in an
operational block 642. The light beam at the emission surface 22
has one or more wavelengths in a range of about 630 nanometers to
about 1064 nanometers, a cross-sectional area greater than about 2
cm.sup.2, and a time-averaged irradiance in a range of about 10
mW/cm.sup.2 to about 10 W/cm.sup.2 across the cross-sectional area,
as described more fully above. The method 640 further comprises
removing heat from the emission surface 22 at a rate in a range of
about 0.1 Watt to about 5 Watts in an operational block 644. The
method 640 further comprises impinging the irradiated surface with
the light beam in an operational block 646.
[0235] The method 640 of certain embodiments further comprises
placing the emission surface 22 in thermal communication with the
irradiated surface (e.g., using the emission surface 22 to apply
pressure to the irradiated surface by applying a force to the
emission surface 22 in a direction generally towards the irradiated
surface, the pressure greater than about 0.1 pound per square inch
or about equal to 2 pounds per square inch).
[0236] In certain embodiments, impinging the irradiated surface
with the light beam is performed for a time period of 10 seconds to
two hours, for a time period of 60 seconds to 600 seconds, or for a
time period of about 120 seconds. In certain embodiments, the steps
of the operational blocks 642, 644, and 646 are performed
concurrently. The method 640 of certain embodiments further
comprises moving the emission surface 22 from a first position at
which a first portion of the irradiated surface is impinged by the
light beam to a second position, and repeating the steps of the
operational blocks 642, 644, and 646 so as to impinge a second
portion of the irradiated surface by light emitted from the
emission surface 22. The first portion and the second portion do
not overlap one another in certain embodiments. This method can be
repeated so as to impinge twenty portions of the irradiated surface
by light emitted from the emission surface 22. In certain such
embodiments, the twenty portions of the irradiated surface do not
overlap one another. However, the portions of the patient's brain
irradiated by impinging these twenty portions of the patient's
scalp do overlap one another in certain embodiments.
[0237] The irradiated surface of certain embodiments of the methods
described above in reference to FIGS. 25-28 comprises a portion of
the patient's scalp or skull. In certain other embodiments, the
surface irradiated by the light comprises a portion of a
light-detection system configured to measure one or more parameters
of light irradiating the surface (e.g., irradiance, total power,
beam size, beam profile, beam uniformity). In certain such
embodiments, the method further comprises measuring the one or more
parameters of the light from the apparatus 10 impinging the
surface. For example, the light-detection system can comprise a
portion of the apparatus 10 configured to test the light beam
emitted from the emission surface 22 immediately prior to treatment
of the patient. In this way, the light-detection system can be used
to ensure that the light beam applied to the patient's scalp or
skull has the desired treatment parameters.
[0238] In certain embodiments, a patient is treated by identifying
a plurality of treatment sites (e.g., at least about 10) on the
patient's scalp or skull, directing a light beam to each of the
treatment sites, and irradiating each treatment site with the light
beam. As described more fully below, in certain embodiments, the
treatment sites are identified using an apparatus comprising a
plurality of indicators, each of which corresponds to a treatment
site location. In certain such embodiments, the treatment sites are
sequentially irradiated by a light beam from the emission surface.
In certain other embodiments, the treatment sites are instead
identified by other indicia. For example, each of the treatment
sites can be identified by markings made on the scalp, or by
structures placed in proximity to the scalp or skull. Each of the
treatment sites can then be irradiated. In certain embodiments,
each of the treatment sites is irradiated by a light beam from the
emission surface while the emission surface is in contact with the
scalp or skull or in contact with an intervening optically
transmissive element which contacts the scalp or skull. In certain
other embodiments, the scalp or skull is not contacted by either
the emission surface or an intervening element. In certain
embodiments, each of the treatment sites is irradiated using a
single beam delivery apparatus which is sequentially moved from one
treatment site to another. In certain other embodiments, a
plurality of beam delivery apparatuses are used to irradiate
multiple treatment sites concurrently. In certain such embodiments,
the number of beam delivery apparatuses is fewer than the number of
treatments sites, and the plurality of beam delivery apparatuses
are sequentially moved to sequentially irradiate the treatment
sites.
[0239] FIG. 29 is a flow diagram of an example method 700 for
controllably exposing at least one predetermined area of a
patient's scalp or skull to laser light to irradiate the patient's
brain. As described more fully below, the method 700 is described
by referring to the wearable apparatus 500 and the beam delivery
apparatus 10 described herein. Other configurations of a wearable
apparatus 500 and a beam delivery apparatus 10 are also compatible
with the method 700 in accordance with embodiments described
herein.
[0240] The method 700 comprises providing a beam delivery apparatus
10 in an operational block 710. In certain embodiments, the beam
delivery apparatus 10 comprises an emission surface 22 configured
to emit a light beam. Other configurations of the beam delivery
apparatus 10 besides those described above are also compatible with
certain embodiments described herein.
[0241] The method 700 further comprises placing a wearable
apparatus 500 over the patient's scalp in an operational block 720.
The apparatus 500 comprises a body 510 and a plurality of
indicators 520. In certain embodiments, each indicator 520 is
substantially transmissive to the light beam emitted from the
emission surface 22. Other configurations of the wearable apparatus
500 besides those described above are also compatible with certain
embodiments described herein.
[0242] The method 700 further comprises placing the emission
surface 22 in thermal communication with a treatment site of the
patient's scalp or skull to be irradiated in an operational block
730. The method 700 further comprises irradiating the treatment
site with light emitted by the emission surface 22 in an
operational block 740. In certain embodiments, the light beam is
transmitted through the indicator 520.
[0243] In certain embodiments, providing the light emitting
apparatus 600 in the operational block 710 comprises preparing the
beam delivery apparatus 10 for use to treat the patient. In certain
embodiments, preparing the beam delivery apparatus 10 comprises
cleaning the portion of the beam delivery apparatus 10 through
which laser light is outputted. In certain embodiments, preparing
the beam delivery apparatus 10 comprises verifying a power
calibration of laser light outputted from the beam delivery
apparatus 10. Such verification can comprise measuring the light
intensity output from the beam delivery apparatus 10 and comparing
the measured intensity to an expected intensity level.
[0244] In certain embodiments, placing the wearable apparatus 500
over the patient's scalp in the operational block 720 comprises
preparing the patient's scalp for treatment. For example, in
certain embodiments, preparing the patient's scalp for treatment
comprises removing hair from the predetermined areas of the
patient's scalp to be irradiated. Removing the hair (e.g., by
shaving) advantageously reduces heating of the patient's scalp by
hair which absorbs laser light from the beam delivery apparatus 10.
In certain embodiments, placing the wearable apparatus 500 over the
patient's scalp in the operational block 720 comprises positioning
the wearable apparatus 500 so that each indicator 520 is in
position to indicate a corresponding portion of the patient's scalp
or skull to be irradiated.
[0245] In certain embodiments, placing the emission surface 22 in
thermal communication with the treatment site in the operational
block 730 comprises pressing the emission surface 22 to the
treatment site. In certain embodiments, by pressing the emission
surface 22 against the treatment site in this way, pressure is
applied to the portion of the patient's scalp of the treatment site
so as to advantageously blanch the portion of the patient's scalp
to be irradiated.
[0246] In certain embodiments, irradiating the treatment site of
the patient's scalp or skull in the operational block 740 comprises
triggering the emission of light from the emission surface 22 by
pressing the emission surface 22 against the treatment site with a
predetermined level of pressure. In certain embodiments, the
emission of light from the emission surface 22 continues only if a
predetermined level of pressure is maintained by pressing the
emission surface 22 against the treatment site. In certain
embodiments, light is emitted from the emission surface 22 to the
treatment site for a predetermined period of time.
[0247] In certain embodiments, the method further comprises
irradiating additional treatment sites of the patient's scalp or
skull during a treatment process. For example, after irradiating a
first treatment site corresponding to a first indicator, as
described above, the emission surface 22 can be placed in contact
with a second indicator corresponding to a second treatment site
and irradiating the second treatment site with light emitted by the
emission surface 22. The various treatment sites of the patient's
scalp or skull can be irradiated sequentially to one another in a
predetermined sequence. In certain embodiments, the predetermined
sequence is represented by the indicators of the wearable apparatus
500. In certain such embodiments, the beam delivery apparatus 10
comprises an interlock system which interfaces with the indicators
of the wearable apparatus 500 to prevent the various treatment
sites from being irradiated out of the predetermined sequence.
[0248] FIG. 30 is a flow diagram of another example method 800 for
treating a patient's brain. The method 800 is described below by
referring to the wearable apparatus 500 and the beam delivery
apparatus 10 described herein. Other configurations of a wearable
apparatus 500 and a beam delivery apparatus 10 are also compatible
with the method 700 in accordance with embodiments described
herein.
[0249] The method 800 comprises noninvasively irradiating a first
area of at least 1 cm.sup.2 of the patient's scalp or skull with
laser light during a first time period in an operational block 810.
The method 800 further comprises noninvasively irradiating a second
area of at least 1 cm.sup.2 of the patient's scalp or skull with
laser light during a second time period in an operational block
820. The first area and the second area do not overlap one another,
and the first time period and the second time period do not overlap
one another. In certain embodiments, the first area and the second
area are spaced from one another by at least 10 millimeters. In
certain embodiments, the first area is over a first hemisphere of
the brain, and the second area is over a second hemisphere of the
brain.
[0250] In certain embodiments, the method 800 further comprises
identifying the first area and the second area by placing a
template over the patient's scalp. The template comprises a first
indicator of the first area and a second indicator of the second
area. For example, the first indicator can comprise a first opening
in the template and the second indicator can comprise a second
opening in the template. In certain embodiments, the method 800
further comprises placing a laser light source at a first position
to noninvasively irradiate the first area and moving the laser
light source to a second position to noninvasively irradiate the
second area.
[0251] In certain embodiments, the method 800 further comprises
increasing the transmissivity of the first area to the laser light
and increasing the transmissivity of the second area to the laser
light. Increasing the transmissivity of the first area can comprise
applying pressure to the first area to at least partially blanch
the first area, removing hair from the first area prior to
noninvasively irradiating the first area, applying an
index-matching material to the first area, or a combination of two
or more of these measures. Increasing the transmissivity of the
second area can comprise applying pressure to the second area to at
least partially blanch the second area, removing hair from the
second area prior to noninvasively irradiating the second area,
applying an index-matching material to the second area, or a
combination of two or more of these measures.
[0252] FIG. 38 is a flow diagram of an example method 900 for
treating a patient's brain in accordance with certain embodiments
described herein. The method 900 comprises providing a patient in
an operational block 910 whose brain has experienced at least one
neurologic disorder (e.g., Alzheimer's Disease, Parkinson's
Disease, depression) or physical trauma (e.g. an ischemic stroke or
a traumatic brain injury) resulting in a blood flow reduction to at
least some brain cells of the patient. The method 900 further
comprises irradiating at least a portion of the patient's scalp or
skull with a pulsed light beam comprising a plurality of pulses
transmitted through the patient's skull in an operational block
920. The pulsed light beam has a temporal profile which supports
one or more intercellular or intracellular biological processes
involved in the survival or regeneration of brain cells. For
example, the pulsed light beam of certain embodiments comprises an
average irradiance per pulse and a temporal profile comprising a
temporal pulse width and a duty cycle sufficient to penetrate the
skull to modulate membrane potentials, thereby enhancing cell
survival (e.g. to cause increased neuron survival), cell function,
or both of the irradiated brain cells.
[0253] In certain embodiments, providing the patient comprises
identifying a patient whose brain has experienced at least one
neurologic disorder or physical trauma. In certain such
embodiments, identifying the patient comprises communicating with
the patient, or with another person with knowledge regarding the
patient's health or experiences, and determining whether the
patient has experienced a neurologic disorder or a physical trauma
to the brain. In certain other embodiments, identifying the patient
comprises examining the patient's body (e.g., head or skull) for
evidence of the patient having experienced a physical trauma to the
brain. This examination in certain embodiments includes use of
invasive or non-invasive medical devices, techniques, or probes
(e.g., a magnetic resonance imaging device). In certain other
embodiments, identifying the patient comprises administering a test
of the patient's mental faculties (e.g., to determine the patient's
abilities on a neurologic function scale) for evidence indicating
that the patient has experienced a neurologic disorder or a
physical trauma to the brain. Persons skilled in the art are able
to identify the patient in accordance with various embodiments
described herein. In certain embodiments, providing the patient
comprises receiving information regarding the results of a previous
identification (e.g., communication, examination, or test
administration) of the patient as one who has experienced at least
one neurologic disorder or physical trauma.
[0254] In certain embodiments, irradiating at least a portion of
the patient's scalp or skull with a pulsed light beam comprises
generating the pulsed light beam and directing the pulsed light
beam to irradiate at least a portion of the patient's scalp or
skull. The pulsed light beam of certain embodiments has a
wavelength, time-averaged irradiance, beam size, beam profile,
divergence, temporal pulse width, duty cycle, repetition rate, and
peak irradiance per pulse, as described herein. Various light
delivery apparatuses can be used to generate the pulsed light beam
and to direct the pulsed light beam towards the patient's scalp or
skull, including but not limited to, the apparatus disclosed herein
or by U.S. Pat. Nos. 7,303,578, 6,214,035, 6,267,780, 6,273,905,
and 6,290,714, and U.S. Patent Appl. Publ. Nos. 2007/0179570 A1,
2007/0179571 A1, and 2005/0107851 A1, each of which is incorporated
in its entirety by reference herein.
[0255] In certain embodiments, irradiating at least a portion of
the patient's scalp or skull comprises identifying one or more
treatment sites (e.g., at least 10, between 2 and 100, or between
15 and 25) and sequentially irradiating the treatment sites with
the pulsed light beam. In certain embodiments, the one or more
treatment sites are identified as described herein (e.g., by an
apparatus worn by the patient and comprising one or more apertures,
by markings made on the scalp, or by structures placed in proximity
to the scalp or skull). In certain embodiments, each treatment site
is irradiated by an apparatus in contact with the scalp or skull or
not in contact with the scalp or skull as described herein. In
certain such embodiments, the irradiated portion of the scalp is
blanched during the irradiation, is not blanched during the
irradiation, is cooled during the irradiation, or is not cooled
during the irradiation.
[0256] In certain embodiments, the patient's scalp is prepared for
treatment prior to irradiation. For example, in certain
embodiments, preparing the patient's scalp for treatment comprises
removing at least a portion of the hair or substantially all the
hair from the predetermined areas of the patient's scalp to be
irradiated. Removing the hair (e.g., by shaving so that the
irradiated portion of the scalp is substantially free of hair)
advantageously reduces heating of the patient's scalp by hair which
absorbs the light from the light emitting apparatus. In certain
other embodiments, the hair is not shaved or otherwise removed
prior to irradiation. For example, irradiating the patient's scalp
can be performed using pulsed light with wavelengths, temporal
pulse widths, and duty cycles which avoid adverse heating of the
patient's scalp due to absorption of light by the hair.
[0257] In certain embodiments, the parameters of the pulsed light
beam used to irradiate the patient's scalp or skull are selected to
perform one or more of the following: (i) to cause increased neuron
survival of the brain cells following at least one physical trauma,
(ii) to support one or more intercellular or intracellular
biological processes involved in the survival or regeneration of
brain cells, or (iii) to modulate membrane potentials in order to
enhance, restore, or promote cell survival, cell function, or both
of the irradiated brain cells following a traumatic brain injury.
In one example such embodiment, the pulsed light beam at the
emission surface of the apparatus has a beam diameter in a range
between 10 millimeters and 40 millimeters, an average irradiance
per pulse in a range between 10 mW/cm.sup.2 and 10 W/cm.sup.2, one
or more wavelengths in a range between 780 nanometers and 840
nanometers, and a temporal pulsewidth in a range between 0.1
millisecond and 150 seconds or between 0.1 millisecond and 300
milliseconds. The duty cycle of certain embodiments can be in a
range between 10% and 30%. Other ranges of these parameters of the
pulsed light beam can be selected in accordance with various other
embodiments described herein.
Neurologic Function Scales
[0258] Neurologic function scales can be used to quantify or
otherwise characterize the efficacy of various embodiments
described herein. Neurologic function scales generally use a number
of levels or points, each point corresponding to an aspect of the
patient's condition. The number of points for a patient can be used
to quantify the patient's condition, and improvements in the
patient's condition can be expressed by changes of the number of
points. One example neurologic function scale is the National
Institute of Health Stroke Scale (NIHSS) which can be used for
short-term measurements of efficacy (e.g. at 24 hours). The NIHSS
is a comprehensive and objective scale which utilizes a
seven-minute physical exam, a 13 item scale, and 42 points. Zero
points corresponds to a normal exam, 42 points (the maximum)
corresponds to basically comatose, and over 15-20 points indicates
that the effects of the stroke are particularly severe. The NIHSS
has previously been used for tPA trials in the treatment of
ischemic stroke, with a 4-point change over 24 hours and an overall
score of 0 or 1 at three months indicative of a favorable outcome.
Other neurologic function scales include, but are not limited to,
modified Rankin Scale (mRS), Barthel Index (BI), Glasgow Outcome,
Glasgow Coma Scale, Canadian Neurologic Scale, and stroke impact
scales such as SIS-3 and SIS-16. In some scales, an improvement in
the patient's condition is indicated by a reduction in the number
of points. For example, the mRS has six points total, with zero
corresponding to normal functioning, and six corresponding to
death. In other scales, an improvement in the patient's condition
is indicated by an increase in the number of points. For example,
in the Glasgow Outcome which has five points, zero corresponds to
death and five corresponds to full recovery. In certain
embodiments, two or more of the neurologic function scales can be
used in combination with one another, and can provide longer-term
measurements of efficacy (e.g., at three months).
[0259] For stroke, the U.S. Food and Drug Administration (FDA) and
the neurologic community have expressed interest in clinical
patient outcomes at 90 days post stroke. Two of the most common and
accepted instruments for measuring efficacy are the NIHSS and mRS.
The FDA is flexible in the way that neurologic function scales can
be used. For example, it is acceptable to use the mRS (i) in
dichotomized fashion with success at score of 0-1 or (ii) it can be
analyzed looking at shifts in the scale showing improvement of
patients along the five-point scale.
[0260] In certain embodiments described herein, a patient
exhibiting symptoms of an ischemic stroke is treated by irradiating
a plurality of treatment sites on the patient's scalp. The
irradiation is performed utilizing irradiation parameters (e.g.,
wavelength, irradiance, time period of irradiation, etc.) which,
when applied to members of a treated group of patients, produce at
least a 2% average difference between the treated group and a
placebo group on at least one neurologic function scale analyzed in
dichotomized or any other fashion and selected from the group
consisting of: NIHSS, mRS, BI, Glasgow Outcome, Glasgow Coma Scale,
Canadian Neurologic Scale, SIS-3, and SIS-16. Certain other
embodiments produce at least a 4% average difference, at least a 6%
average difference, or at least a 10% average difference between
treated and placebo groups on at least one of the neurologic
function scales analyzed in dichotomized or any other fashion and
selected from the group consisting of: NIHSS, mRS, BI, Glasgow
Outcome, Glasgow Coma Scale, Canadian Neurologic Scale, SIS-3, and
SIS-16. In certain embodiments, the irradiation of the patient's
scalp produces a change in the patient's condition. In certain such
embodiments, the change in the patient's condition corresponds to a
change in the number of points indicative of the patient's
condition. In certain such embodiments, the irradiation produces a
change of one point, a change of two points, a change of three
points, or a change of more than three points on a neurologic
function scale.
Transmission in Human Brain
[0261] Power density (PD) measurements have been made to determine
the transmission of laser light having a wavelength of
approximately 808 nanometers through successive layers of human
brain tissue. Laser light having a wavelength of (808.+-.5)
nanometers with a maximum output of approximately 35 Watts was
applied to the surface of the cortex using a beam delivery system
which approximated the beam profile after the laser light passes
through the human skull. Peak power density measurements were taken
through sections of human brain tissue using an Ocean Optics
spectrophotometer Model USB 2000, Serial No. G1965 and beam
diameter after scattering was approximated using a Sony Model
DCR-IP220, Serial No. 132289.
[0262] A fresh human brain and spinal cord specimen (obtained
within six hours after death) was collected and placed in
physiologic Dakins solution. The pia layer, arachnoid layer, and
vasculature were intact. The brain was sectioned in the midline
sagittaly and the section was placed in a container and
measurements taken at thicknesses of 4.0 centimeters (.+-.0.5
centimeter), 2.5 centimeters (.+-.0.3 centimeter), and 1.5
centimeters (.+-.0.2 centimeter). The PD measurements are shown in
Table 1:
TABLE-US-00001 TABLE 1 Thickness PD at cortex Average PD at
thickness 4.0 cm 20 mW/cm.sup.2 4.9 .mu.W/cm.sup.2 2.5 cm 20
mW/cm.sup.2 20 .mu.W/cm.sup.2 1.5 cm 10 mW/cm.sup.2 148
.mu.W/cm.sup.2
FIG. 31 is a graph of the PD versus the depth from the dura for an
input PD of 10 mW/cm.sup.2 with the light bars corresponding to
predicted values of the PD and dark bars corresponding to an
estimated minimum working PD of is 7.5 .mu.W/cm.sup.2, as described
below.
[0263] Based upon prior animal experimentation, a conservative
estimation of the minimum known PD within the tissue of the brain
which is able to show efficacy in stroke animal models is 7.5
.mu.W/cm.sup.2. This estimated minimum working PD is drawn from an
experiment in which 10 mW was applied to the rat brain surface, and
7.5 .mu.W/cm.sup.2 PD was directly measured 1.8 centimeters from
the surface. This stroke model consistently produced significant
efficacy, including for strokes 1.8 centimeters from the laser
probe. Note that this 7.5 .mu.W/cm.sup.2 is a conservative
estimate; the same irradiance or power density at the brain surface
also consistently produces significant efficacy in a 3-centimeter
rabbit clot shower model. Note also that the power density
measurements in the human brain experiment do not factor in the
effect from the CNS-filled sulci, through which the laser energy
should be readily transmitted. However, even conservatively
assuming 7.5 .mu.W/cm.sup.2 as the minimum power density hurdle and
ignoring expected transmission benefits from the sulci, the
experiment described above confirms that approximately 10-15
mW/cm.sup.2 transmitted upon the cortex (as per an example
dosimetry in man) will be effective to at least 3.0 centimeters
from the surface of the brain.
In Vivo Thermal Measurements
[0264] In vivo thermal measurements were made to determine the
heating effect in living tissue of laser light having a wavelength
of approximately 808 nanometers. A GaAlAs laser source of
808-nanometer light was placed in direct contact with the skin of
the heads of live rabbits and rats. The laser source had an
approximately Gaussian beam profile with a beam diameter of 2.5-4.0
millimeters (1/e.sup.2). Thermocouple probes (Model Bat-12 from
Physitemp Instruments Inc. of Clifton, N.J.) were placed in the
subcutaneous tissue and below the dura and measurements were
recorded at various irradiances or power densities. The results of
these measurements are shown in Table 2:
TABLE-US-00002 TABLE 2 Exposure Temperature Animal Probe location
Dose time increase Rat Subcutaneous 15 mW/cm.sup.2 4 minutes
approximately 3.degree. C. Rat Subdural 15 mW/cm.sup.2 4 minutes
approximately 1.degree. C. Rat Subcutaneous 75 mW/cm.sup.2 4
minutes approximately 7.degree. C. Rat Subdural 75 mW/cm.sup.2 4
minutes approximately 7.degree. C. Rabbit Subcutaneous 7.5
mW/cm.sup.2 5 minutes less than 0.5.degree. C. Rabbit Subdural 7.5
mW/cm.sup.2 5 minutes less than 0.5.degree. C. Rabbit Subcutaneous
37.5 mW/cm.sup.2 5 minutes approximately 5.5.degree. C. Rabbit
Subdural 37.5 mW/cm.sup.2 5 minutes less than 0.5.degree. C.
[0265] There is minimal heating (e.g., less than 0.5.degree. C.) in
the subdural region at four times the therapeutic energy density.
The "heat sink" effect of living tissue that minimizes possible
heating in the cortex is significantly larger in humans than in
rats or rabbits, due to the larger heat sink and blood flow volume,
which further limits the undesirable effects of heating in the
region of stroke. Therefore, in certain embodiments described
herein, a therapeutic dosage of energy is delivered to the area of
a stroke without undesirable heating of the dura.
Phototherapy Example 1
[0266] One example of phototherapy (Lampl Y, Zivin J A, Fisher M,
Lew R. Welin L, Dahlof B, Borenstein P, Andersson B, Perez 1,
Caparo C, Ilic S, Oron U. Infrared laser therapy for ischemic
stroke: a new treatment strategy: Results of the NeuroThera
Effectiveness and Safety Trial-1 (NEST-1). Stroke. 2007;
38:1843-1849, incorporated in its entirety by reference herein,
suggested the safety and efficacy of transcranial light therapy
(TLT) for treatment of humans 40 to 85 years of age with ischemic
stroke within 24 hours of stroke onset in a small randomized,
controlled trial. The NeuroThera Laser System therapeutic approach
involves use of infrared laser technology and has shown significant
and sustained beneficial effects in animal models of ischemic
stroke.
[0267] The NeuroThera Laser System (NTS) used in this NEST-1 study
utilized an infrared laser technology that involves
photobiostimulation. A large and growing body of scientific
literature is available documenting the photobiostimulation effects
of infrared laser therapy both in vitro and in vivo. The biological
effects of infrared laser therapy are wavelength-specific and are
not attributable to thermal effects. Energy in this region of the
electromagnetic spectrum is nonionizing and, therefore, poses none
of the hazards associated with UV light. It has been demonstrated
that irradiation of specific infrared wavelengths is able to
penetrate deeply into the brain. This form of therapy is
distinguished from photodynamic therapy, which involves using light
energy to penetrate the body and to activate a photosensitive
drug.
[0268] Photobiostimulation involves increased adenosine
triphosphate (ATP) formation after energy absorption inside
mitochondria. A compound that absorbs energy in the spectral region
of interest is known as a chromophore. There is evidence that
suggests that a primary mitochondrial chromophore for
photobiostimulation is cytochrome c oxidase. This enzyme complex
contains 2 copper centers, CU.sub.A and CU.sub.B. The primary
chromophore for the NTS wavelength is in the CU.sub.A center which
has a broad absorption peak around 830 nm in its oxidized form. The
NTS delivers energy at 808 nm, which is within this absorption
peak, and is able to penetrate into the brain noninvasively.
Cytochrome c oxidase is a terminal enzyme in the cellular
respiratory chain and is located in the inner mitochondrial
membrane. It plays a central role in the bioenergetics of
eukaryotic cells by delivering protons across the inner membrane,
and thereby driving the formation of ATP by oxidative
phosphorylation. In addition to leading to increased ATP formation,
photobiostimulation may also initiate secondary cell-signaling
pathways. The overall result is improved energy metabolism,
enhanced cell viability, and may also involve prevention of
apoptosis in the ischemic penumbra and enhancement of neurorecovery
mechanisms.
[0269] In vivo studies have suggested that infrared laser therapy
could be beneficial for the treatment of acute myocardial
infarction, acute ischemic stroke, injured peripheral nerves and
spinal cord injury. Previous studies have shown in 2 different
animal models a positive impact of infrared laser therapy on the
experimental, ischemic stroke treatment outcomes in New Zealand
rabbits (rabbit small clot embolic stroke model [RSCEM]) and
Sprague-Dawley rats (permanent middle cerebral artery occlusion).
Lapchak has shown that laser treatment at 6 hours poststroke onset
in RSCEM improved behavioral performance and produced a durable
effect that was measurable 21 days after embolization. De Taboada
and Oron have also shown that laser treatment up to 24 hours
poststroke onset in permanent middle cerebral artery occlusion
showed significant improvement in neurological deficits which was
evident at 14, 21 and 28 days poststroke when compared with the
sham control group. Currently, the putative mechanism for infrared
laser therapy in stroke involves the stimulation of mitochondria,
which then leads to preservation of tissue in the ischemic penumbra
and enhanced neurorecovery. The exact mechanistic pathways remain
to be elucidated.
Study Design
[0270] NEST-1 was a prospective, multicenter, international,
double-blind, randomized, sham (placebo) controlled trial conducted
at 6 medical centers in 3 countries: Israel, Peru, and Sweden. The
study examined initial safety and effectiveness of infrared
wavelength laser therapy for treatment of patients within 24 hours
of ischemic stroke onset.
[0271] This study was conducted in accordance with the FDA/ICH Good
Clinical Practice guidelines and applicable local regulatory
requirements. Investigators were required to ensure that this study
was conducted in full conformity with the 1983 revision of the
Declaration of Helsinki or with the laws and current regulations in
biomedical research involving human patients of the country in
which the study was conducted, whichever afforded greater
protection to the patients. The protocol and information for
patients and healthcare providers was approved by each center's
ethics committee or Institutional Review Board. Country-specific
independent data monitoring committees conducted safety reviews
throughout the study.
[0272] Eligible patients were required to be between 40 to 85 years
of age, have a clinical diagnosis (within 24 hours of stroke onset)
of ischemic stroke causing a measurable neurological deficit (total
NIHSS score ranging from 7 to 22 at admittance to the medical
center), and to have NTS treatment initiated within 24 hours from
stroke onset. The patient or parent legal representative gave
written informed consent before enrollment into the study.
Ineligibility Criteria
[0273] Patients were excluded if: there was evidence on a CT scan
of an intracranial, subdural or subarachnoid hemorrhage, or
clinical presentation suggestive of subarachnoid hemorrhage, even
if the initial CT scan was normal; the patient was a candidate for
intravenous or intra-arterial administration of tissue-type
plasminogen activator or other thrombolytic therapy for treatment
of the acute ischemic stroke, and tissue plasminogen activator or
other thrombolytic therapy was administered; the patient had a
seizure at stroke onset; serum blood glucose was >400 mg/dL (22
mmol/L) or <40 mg/dL (2.2 mmol/L); the patient had sustained
hypertension (defined during the baseline period by 2 readings
occurring 30 minutes apart with systolic blood pressure >185
mmHg or diastolic blood pressure >110 mmHg) at time of treatment
or need for aggressive treatment for blood pressure reduction;
there was sustained hypotension (defined as systolic blood pressure
<80 mmHg, or diastolic blood pressure <50 mmHg); there was
presumed septic embolus; the patient had known hereditary or
acquired hemorrhagic diathesis, e.g., activated partial
thromboplastin time or prothrombin time greater than normal,
unsupported coagulation factor deficiency, or oral anticoagulant
therapy with the prothrombin time greater than normal; the patient
had a skin condition (i.e., hemangioma, scleroderma, psoriasis,
rash, or open wound) at the site chosen for infrared energy
application; the patient was previously enrolled in or had
participated in another investigational drug or device trial within
the preceding 4 weeks; if a new medication was started within 14
days before the screening visit; the participant had severe mental
deficit, severe neurological deficit or disorder (dementia,
multi-infarct dementia, advanced multiple sclerosis) which would
interfere with the assessment of the patient's ability for
independent functioning; there was evidence of any disorder other
than stroke that, in the opinion of the investigator, could be
considered serious or life threatening such as active serious
infections, pneumonia, pulmonary emboli, or gastrointestinal
bleeding; the patient had unstable cardiac arrhythmias or other
cardiac illness that, in the opinion of the investigator, was life
threatening; the patient was of child bearing potential; the
patient was comatose or moribund level of consciousness; or the
patient was otherwise determined by the investigator to be
medically unsuitable for participation in this study.
Study Groups, Evaluation Measures, and Baseline Factors
[0274] All patients received standard medical management therapy
for acute ischemic stroke. In addition, they all underwent an
identical NTS procedure. A randomization code that was
preprogrammed within the NeuroThera Laser System determined whether
the treatment was active or sham (placebo). Both patients and
clinicians were blinded regarding treatment arm. The National
Institutes of Health Stroke Scale (NIHSS) was assessed at the time
of screening for entry into the study and again immediately before
randomization to treatment group. Outcome measures (NIHSS, modified
Rankin Scale [mRS], Barthel Index, and Glasgow Outcome Scale) were
determined at 30, 60, and 90 days. Neurological scores and clinical
data were collected on standard case report forms at each visit by
trained investigators.
[0275] Baseline factors including patient demographics, time to
treatment, medical history, vital signs, and routine laboratory
values were collected. Factors also included age, sex, time from
stroke onset to arrival at hospital, time from stroke onset to
treatment, and a complete medical history.
[0276] After completion of the NTS procedure, patients entered the
study follow-up phase until one of the following occurred: the
patient decided to stop participation in the study; the sponsor or
ethics committee/applicable regulatory body terminated the study;
the investigator decided to discontinue the patient or site
participation in the study; or the patient had participated in the
study for 90.+-.10 days.
The NTS Treatment Device
[0277] The NTS used in the NEST-1 study was an investigational
device intended to provide noninvasive, transcranial laser
treatment to patients diagnosed with acute ischemic stroke. The
laser wavelength of 808 nm is in the near-infrared portion of the
electromagnetic spectrum, and is invisible to the naked eye. Energy
in the near-infrared spectrum is nonionizing and is not associated
with the risks of ionizing radiation. The NTS device used in the
NEST-1 study included a class IV laser system and delivers energy
via a fiber optic cable to a handheld probe that is placed on the
shaved head of the patient by a trained operator. The device is
portable and is similar in size to portable ultrasound
equipment.
[0278] The NTS is manufactured by PhotoThera, Inc. A complete
treatment regimen as defined for the NEST-1 study included removing
hair from the patient's scalp, followed by NTS application (active
treatment or sham/control treatment) on 20 predetermined locations
on the scalp for 2 minutes at each site. The predetermined sites
are identified by a cap which is placed on the patients head. The
system is designed to deliver about 1 Joule/cm.sup.2 of energy over
the entire surface of the cortex regardless of stroke location. The
sham procedure is identical to the active procedure with the
exception that no laser energy is delivered to the patient from the
device.
[0279] Based on current knowledge of the technology and risk
assessment analysis, the most significant known hazard with NTS
treatment is potential retinal damage if the beam enters through
the lens of the eye and onto the retina. Other potential hazards
include skin burns and cuts to the scalp from shaving the head.
Skin burns could occur if the device is not used as intended (eg,
repeated treatments at the same location).
Statistical Methods
[0280] Effectiveness outcomes were reported on an
intention-to-treat basis and include all 120 patients randomized to
both arms. Safety outcomes were based on the same 120 patients, who
also comprised all patients who received any treatment. Patients
were evaluated at baseline, 30, 60, and 90 days after baseline.
Analysis focused largely on the 90-day evaluations. The NIHSS was
the prospectively identified primary outcome, and the mRS, Glasgow
Outcome Scale and Barthel Index scores were secondary outcomes.
[0281] Categories of baseline values of the NIHSS score and of time
from stroke onset to treatment were entered into the analyses as
strata or covariates. The three NIHSS strata were 7 to 10, 11 to
15, and 16 to 22. The categories for time from stroke onset to
treatment were "less than 12 hours" and "12 to 24 hours." The NIHSS
scale is not an interval scale. Therefore, categories of the NIHSS
score were used to reduce potential heterogeneity.
[0282] NIHSS outcome was collapsed into a binary outcome, bNIH,
where success' could occur in either of 2 ways: as a 90-day NIHSS
score 0 to 1 or as a decrease in score (change) of 9 or more points
from baseline to 90 days.
[0283] The mRS 90-day outcome took 2 forms. The 7-category ordinal
variable form, analyzed across the whole distribution of scores on
the 0 to 6 mRS scale (full mRS), and a binary mRS that makes scores
of 0 to 2 as positive (success) and scores of 3 to 6 as negative
(failure).
[0284] The full mRS ("shift in Rankin"), binary mRS and bNIH
outcomes were tested using a stratified Cochran-Mantel-Haentzel
(CMH) test: namely, the van Elteren test. The test uses the
modified ridit score and thereby is a direct extension of the
2-sample Wilcoxon test. For the bNIH and the binary mRS outcomes,
logistic regression analyses were used to explore the effects of
covariates and the random effect of site: in particular, to assess
how adding these factors altered the estimate of treatment
effect.
[0285] The analyses were carried out in SAS version 9 using PROC
FREQ to obtain the results for the van Elteren CMH test, and using
PROC GENMOD and PROC LOGISIC to obtain results for logistic
regression analyses with and without medical center as a random
effect. Prevalence odds ratios were obtained from PROC GENMOD.
[0286] This study was an exploratory trial rather than a
confirmatory trial, in the sense of FDA/ICH E8 Guidance on General
Considerations for Clinical Trials and FDA/ICH E9 Guidance on
Statistical Principles for Clinical Trials. Primary safety and
effectiveness outcome measures and their analysis were identified
prospectively. Multiple secondary and exploratory analyses were
defined in the protocol or were designed and performed after study
completion and unblinding. No corrections were made for multiple
comparisons.
[0287] The study enrolled 122 eligible adult patients, between ages
40 and 85 of any ethnic background diagnosed with acute ischemic
stroke within 24 hours of onset who provided their written informed
consent. Two patients withdrew before randomization and are not
included in any analyses, leaving 120 patients in the effectiveness
analysis. Of the 120 patients, 79 were randomized to the active
treatment group and 41 were randomized to the sham control group.
FIG. 32 shows the disposition of patients in the study. There was
only patient lost to follow-up (0.8%). No significant differences
in baseline characteristics were observed (see Table 3; baseline
demographics and other baseline characteristics). Study data were
reviewed by independent data monitoring committees in each country;
there were no serious device-related adverse effects reported.
TABLE-US-00003 TABLE 3 Active Treatment (NTS), Placebo (sham
Characteristic n = 79 control), n = 41 Mean age, y 70.2 68.5
Female, No. (%) 36 (45.6%) 15 (36.6%) Ethnicity, No. (%) White 29
(36.7%) 17 (41.5%) Black 2 (2.5%) 0 (0.0%) Hispanic 4 (5.1%) 2
(4.9%0 Other (largely Mestizo and 44 (55.7%) 22 (53.7%) Native
American Indians) Median time to treatment, h 18 17 Mean time to
treatment, 16:56 16:20 hr:min Mimimum, hr:min 02:00 04:05 Maximum,
hr:min 23:56 23:22 Median NIHSS score at entry 11 10 First quartile
9 9 Third quartile 15 14 History, No. (%) Hypertension 44 (55.7%)
20 (48.8%) Previous stroke 17 (21.5%) 12 (29.3%) Diabetes mellitus
20 (25.3%) 9 (22.0%)
Effectiveness Analysis
[0288] The proportion of patients who received active treatment and
had a positive bNIH outcome was 70%, which is greater than the
proportion who received sham control treatment with a positive bNIH
outcome (51%; CMH test P=0.035 stratified by severity and time from
stroke onset to treatment; P=0.048 stratified only by severity).
The treatment effect remained significant with other choices of
strata for the CMH analysis. Logistic regression analyses confirmed
that the results held controlling for both fixed covariates (eg,
age, sex, time-to-treatment, baseline severity, previous stroke)
and the random effects of medical site. Controlling only for
baseline severity the logistic regression gave a prevalence odds
ratio favoring treatment of 1.40 (95% CI, 1.01 to 1.93). Among the
79 treated patients, 38% achieved both a final NIHSS score of 0 to
1 and improved by .gtoreq.9 points, 20% had only a .gtoreq.9-point
improvement, 11% obtained a final score of 0 to 1 without improving
by .gtoreq.9, and 30% achieved neither end point. Among the 41
control patients the corresponding proportions were 29%, 7%, 15%,
and 49%.
[0289] Differences in mean NIHSS scores between the treatment
groups appeared soon after treatment and were apparent throughout
the 90-day study period. FIG. 33 shows the mean NIHSS over time for
each treatment group. Patients in the active treatment group showed
greater improvement in the change in NIHSS scores from baseline to
day 90, as compared with the sham control group (P=0.021, CMH test
stratified by time to treatment).
[0290] For the binary mRS outcome (0 to 2 versus 3 to 6), a similar
pattern of significance held. The proportion of patients who
received active treatment and had a positive binary mRS outcome was
60%, which is greater than the proportion who received sham control
treatment with a positive binary mRS outcome (44%; CM test P=0.034
stratified by severity and time to treatment; P=0.043 stratified
only by severity). Only the CMH test without strata was not
significant (P<0.11 X.sup.2 test). The rate of positive results
markedly varies across the baseline severity strata. Controlling
only for baseline severity, logistic regression gave prevalence
odds ratios favoring treatment of 1.38 (95% CI, 1.03 to 1.83) for
the binary mRS outcome.
[0291] The effect of the NTS when compared with sham treatments
with respect to the score on the full mRS at 90 days or the last
rating, analyzed across the whole distribution of scores on the 0
to 6 mRS scale was significant, with the use of the
Cochran-Mantel-Haentzel nonparametric rank test, stratified by
categories of (1) baseline NIHSS score and time to treatment
(P=0.020) and (2) baseline NIHSS score only (P=0.026; see Table
4).
TABLE-US-00004 TABLE 4 mRS 0 1 2 3 4 5 6 Total Control 5 10 3 8 6 5
4 41 % 12.20 24.39 7.32 19.51 14.63 12.20 9.75 Active 12 25 10 11
12 2 7 79 % 15.19 31.65 12.66 13.92 15.19 2.53 8.86 Total 17 35 13
19 18 7 11 120
[0292] FIG. 34 shows the mean nRS over time for each treatment
group. Stratification by baseline severity gave similar results for
the 3 outcomes (bNIH, binary mRS, and full mRS); all 3 outcomes had
significance levels <0.05 (see Table 5; 2-sided significance
levels for the van Elteren CMH test). When also controlled for
time-to-treatment (0 to 12 hours versus 12 to 24 hours) little
significance is gained. However, in a trial with a larger sample
size, time-to-treatment would be expected to have a stronger
association with outcome.
TABLE-US-00005 TABLE 5 Stratified Stratified by Severity and
Outcome by Severity Only Time-to-Treatment Binary_NIH 0.048 0.035
Binary_mRS 0.043 0.034 Full mRS 0.026 0.020
[0293] Results of analyses using the Glasgow Outcome Scale and
Barthel Index are similar to those for the NIHSS and the mRS.
Patients who received active treatment had better outcomes than
patients who received sham control treatment as measured on the
Glasgow Outcome Scale (CMH test P=0.056), and the Barthel Index
scale (CMH test P=0.035), stratified by baseline NIHSS score and
time to treatment.
[0294] The logistic regression analyses indicated the negligible
effects of covariate adjustment on the logistic regression
coefficient for treatment. The results in Table 6 indicate that
treatment effect is stable across 2 binary outcomes and across 3
different nested sets of covariates. In fact, the treatment effect
tends to increase as covariates are added. Furthermore, treating
hospital site as a repeated measures effect virtually does not
alter the logistic regression coefficient for treatment. The 95%
CIs are not shown to focus on the consistency of the regression
coefficients. In all but one model the probability value for
treatment is <5%. In the models with the bNIH outcome, the
covariate `severity` is not significant and time-to-treatment is
significant only once with P=0.0496. However with the binary mRS
outcome, the covariate `severity` is significant with P<0.001 in
all 3 models. This indicates that the 9-point decrease in the NIHSS
score captures the variation of treatment effect across the
baseline severity categories. We explored many sets of additional
covariates and found that after including the covariates `gender`,
`age`, and `prior stroke` all other covariates had negligible
predictive value. Gender was significant in the binary mRS model
with P<0.01. Otherwise, these factors did not achieve
statistical significance. See Table 6 which shows the results for
two sets of nested multivariate models, 1 set of models with
outcome bNIH and 1 set of models with outcome binary mRS
(time-to-tx indicates time from onset to treatment). The first P
value is for simple logistic regression and the second P value is
for logistic regression with the factor `site` included as a
repeated measures effect.
TABLE-US-00006 TABLE 6 Covariates in Model Outcome bNIH Outcome
Binary mRS TX Coefficient and TX Coefficient (P values)* and (P
values)* Severity 0.82 (0.046) 0.92 (0.044) 0.82 (0.007) 0.92
(0.095) Severity and time-to-tx 0.89 (0.034) 1.03 (0.027) 0.89
(0.009) 1.03 (0.032) Severity, time-to-tx, 0.99 (0.027) 1.43
(0.009) gender, age, prior stroke 0.99 (0.010) 1.44 (0.020)
Safety Analysis
[0295] Table 7 (mortality rates and SAEs) shows the mortality rates
and SAEs by treatment group and totals. No significant difference
in mortality between the active treatment group and the sham
control group is evident. Table.sub.--7 shows the number of
patients with serious adverse events, worsening of underlying
disease, cardiovascular SAEs, infection, or central nervous system
SAEs, in total and by treatment group. These data indicate that
there were no significant differences between the treatment groups
with respect to these measures. Where there was a trend toward
differences between the treatment groups, such as in rates of
infection or rates of central nervous system SAEs, the patients
receiving active treatment appear to have had better outcomes than
patients receiving sham control treatment.
TABLE-US-00007 TABLE 7 No. of Total, Fisher Patients n = Active,
Sham, Exact, P with: 120 Percent n = 79 Percent n = 41 Percent
Value Mor- 11 9.2% 7 8.9% 4 9.8 0.87 tality (all sites) SAEs 35
29.2% 20 25.3% 15 36.6 0.211 Wors- 8 6.7% 3 3.8% 5 12.2 0.120 ening
of under- lying disease CVS 8 6.7% 5 6.3% 3 7.3 1.000 Infec- 13
10.8% 5 6.3% 8 19.5 0.059 tion CNS 14 11.7% 6 7.6% 8 19.5 0.072
Discussion
[0296] The NEST-1 trial provides initial evidence on the safety and
effectiveness of infrared laser therapy for the treatment of
ischemic stroke in humans within 24 hours of stroke onset. The
outcome variable scales used in the NEST-1 study had excellent
correlation: R=0.79 to 0.92. The correlation coefficients for the
NEST-1 trial are essentially the same as those reported in the
article by Lyden and colleagues reviewing tissue plasminogen
activator data. That is, the outcome variables have correlation
coefficients with each other of about 0.8 (absolute value) or
higher. This concordance with prior studies is evidence that the
outcomes are being measured appropriately and consistently.
[0297] The results suggest that infrared laser therapy may benefit
a broad spectrum of stroke patients without increasing the rate of
adverse events. Furthermore, the relatively large magnitude of the
effect implies that a phase III trial should not require a
substantial number of subjects.
[0298] Patients receiving active treatment had a higher proportion
of positive NIHSS outcomes than did patients receiving sham control
treatment. Results were similar using the other neurological
outcome scales. No significant differences between the treatment
groups were observed in rates of mortality or SAEs, but the sample
size (n=120) gives low power to detect small differences. Where
there is a trend toward differences between the treatment groups,
patients receiving active treatment appeared to have had fewer SAEs
than did patients receiving sham control treatment. The safety
profile of the NTS treatment as demonstrated in this study was
clear. There were no adverse outcomes that can be attributed to the
laser therapeutic procedure.
[0299] The bNIH outcome with the 9-point change incorporates
variation in baseline severity (from NIHSS score 7 to 22) and
suggests a global potential benefit. In contrast, the binary mRS
outcome does not account for change from baseline. Thus, once the
analysis controlled for baseline severity, the results based on the
2 binary outcomes closely agreed. Controlling for baseline
severity, the analyses by the CMH test and by logistic regression
gave prevalence odds ratios favoring treatment exceeding 1.40 for
the bNIH outcome and exceeding 1.38 for the binary mRS outcome.
[0300] This global potential benefit is also demonstrated through
the full mRS, analyzed across the entire distribution of Rankin
scores, from 0 to 6. The mRS is a simple and reliable outcome
measure when consistently implemented by trained clinicians. The
full mRS analysis takes into consideration the entire spectrum of
the patient outcomes. As a result, the full mRS is increasingly
considered as a primary outcome measure for ischemic stroke trials
involving neuroprotective technologies.
[0301] This exploratory study had a prespecified analytic plan with
a primary outcome of bNIH, the binary form of NIHSS that regards a
final score of 0 to 1 or a 9-point decrease as a success. But our
presentation of several analytic approaches raises the concern of
type 1 error. We described several approaches to the same
hypothesis: some having an mRS outcome, some having an NIHSS
outcome, and some using logistic regression to confirm the
nonparametric results. These results showed the substantial
concordance among these outcomes and methods. Also, they showed
that after control for NIHSS baseline severity, other factors had
little or no effect on the magnitude of the treatment effect.
Hence, we did not present a multiple comparisons correction such as
the Bonferroni correction because, in particular, the Bonferroni
correction assumes that the hypotheses are independent of one
another. Another reason for the various analyses was to associate
an effect size with the results of the primary analysis by the
nonparametric CMH test. Simple estimates of effect size were
obtained from the other tests and both the simple proportions of
success for the binary outcomes and prevalence odds ratios obtained
from logistic regression were reported.
[0302] An extended treatment window of up to 24 hours after stroke
onset will have a number of implications. Thrombolytics have a
proven treatment window of 3 hours, although it may be that
effectiveness for this form of therapy extends out somewhat
further. The first neuroprotective trial to show efficacy was the
study of NXY-059. That study had a 6-hour treatment window, but a
majority of patients were treated within 4 hours. It is a
reasonable contention that the reason the NXY-059 study was
successful, whereas all the previous neuroprotective therapies were
not, was that the average time to treatment was kept so low. NEST-1
had a 24-hour treatment window and a much longer time to treatment
(median 18 hours) than nearly all other clinical trials to date for
the treatment of acute ischemic stroke. A major problem for
treatment of strokes has been that large numbers of patients
present after 6 hours. Therefore, an expanded treatment window of
24 hours would make it possible to treat many more ischemic stroke
victims.
[0303] Although the mechanism of action of infrared laser therapy
for stroke is not completely understood, a number of effects of
this type of irradiation have been documented. Infrared laser
therapy is a physical process that can produce biochemical changes
at the tissue level. The putative mechanism for NTS treatment
involves stimulation of ATP formation by mitochondria and may also
involve prevention of apoptosis in the ischemic penumbra and
enhancement of neurorecovery mechanisms. An example of another
physical process that reduces neurological damage is hypothermia.
In animal model studies, there are few, if any therapies that have
been shown as consistently to reduce stroke-related damage as
hypothermia. What is clear is that infrared irradiation is probably
delivering its effect independent of restoration of blood flow and
the mechanism is probably related to an improved energy metabolism
and enhanced cell viability.
[0304] Other advantages of this form of therapy are that treatment
can be started rapidly, without any need for preliminary laboratory
testing, invasive procedures, or extensive training of the
clinicians who administer the treatment. Furthermore, it is not
necessary to know the location of the vascular occlusion to
administer the NTS treatment. Thus, this form of therapy is likely
to require much less infrastructure than virtually all other types
of devices and medical therapies available to date for acute stroke
treatment or clot removal.
[0305] Although the NEST-1 study results are encouraging, and may
indicate that infrared laser therapy has potential to become a
treatment of ischemic stroke in humans when initiated within 24
hours of stroke onset, a larger confirmatory trial to demonstrate
safety and effectiveness is warranted.
Phototherapy Example 2
[0306] Another example of phototherapy (NeuroThera Effectiveness
and Safety Trial-2 (NEST-2) was nearly identical to the trial study
discussed above, but was larger and included patients 40 to 90
years of age. NEST-2 was a double blind, placebo (sham) controlled
trial in which 660 patients were enrolled at 57 centers in 4
countries. Patients were eligible for inclusion in the study if
they were 40 to 90 years of age, had a baseline NIHSS score between
7 to 22, had a clinical diagnosis of ischemic stroke, no evidence
of hemorrhagic infarct by CT scan or MRI, and had not received tPA.
Initiation of treatment had to occur within 24 hours after stroke
onset. The inclusion and exclusion criteria are summarized in Table
8.
TABLE-US-00008 TABLE 8 Major Inclusion and Exclusion Criteria
Inclusion 40-90 years of age Diagnosis of acute ischemic stroke
within 24 hours of onset NIHSS .gtoreq.7-.ltoreq.22 Informed
consent Exclusion Evidence of intracranial, subdural, or
subarachnoid hemorrhage Prestroke .gtoreq.3 mRS Clinical diagnosis
of a brain stem or cerebellar stroke Seizure at onset Blood glucose
>400 or <60 Sustained systolic BP >220 mmHg, <80 mmHg
or diastolic >140 mmHg, <50 mmHg Suspected septic embolus CNS
tumor (except asymptomatic meningioma) Dermatologic conditions
(eg., psoriasis) of the scalp Thrombolytic stroke therapy Head
implant (eg, clipped aneurysm, Hakim valve) Photodynamic therapy
within 14 days (eg, Visudyne) Pregnancy Severe comorbidities CNS
indicates central nervous system.
Randomization and Treatment
[0307] Patients were randomly assigned in a 1:1 ratio to receive
either TLT or sham. All patients underwent the identical TLT
procedure that involves removal of the patient's scalp hair
followed by application of a laser probe to the patient's head. The
total procedure time was approximately 2 hours. After consent, an
interactive voice randomization system was used, with dynamic
randomization at centers to ensure balanced distribution of
treatment assignments. All patients received standard of care
medical management throughout the course of trial.
Study Management
[0308] This study was conducted in accordance with the FDA/ICH Good
Clinical Practice (GCP) guidelines and applicable local regulatory
requirements. The trial was also conducted in full conformity with
the 1983 revision of the Declaration of Helsinki or with the laws
and current regulations in biomedical research involving human
patients of the country in which the study was conducted, whichever
afforded greater protection to the patients. The study was designed
and overseen by the steering committee. Each center's ethics
committee or Institutional Review Board and an independent data
monitoring committee (DMC) conducted safety reviews. The DMC
periodically reviewed serious adverse event data between the
groups. The patient or legal representative gave written informed
consent before enrollment into the study. Data management and
statistical analysis was conducted by an independent contract
research organization (Parexel, Waltham, Mass), as well as by the
study sponsor (PhotoThera, Carlsbad, Calif.). After database lock
and independent review by the DMC, the steering committee had
complete access to the trial data and assumed responsibility for
the analysis and interpretation of the results.
Treatment
[0309] The NeuroThera Laser System (NTS) included an apparatus with
a hand-held housing to apply the light to selected portions of the
patient's scalp. The NTS uses energy at a wavelength of 808 nm
which is near-infrared, nonionizing, and is invisible to the naked
eye. Experiments using cadavers indicated that optimal amounts of
laser energy are able to penetrate the brain to a depth of
approximately 2 cm. The dosimetry of wavelength and irradiance
levels was based on transmission experiments with fresh human
cadavers without fixation. Monte Carlo simulations, in vitro
experiments, review of the literature using infrared therapy for
other indications and the results of preclinical studies in
validated stroke animal models. A complete treatment regimen as
defined for the NEST-2 study included applying the hand-held probe
on 20 predetermined locations on the shaved scalp for 2 minutes at
each site. The predetermined sites, which are irrespective of
stroke location, are identified by a cap which is placed on the
patient's head. The sham procedure is identical to the TLT
procedure with the exception that no laser energy is delivered to
the patient from the device.
[0310] Based on current knowledge of the technology and risk
assessment analysis, the most significant known hazard with TLT is
potential retinal damage if the beam enters the eye and is focused
onto the retina which could result in permanent eye injury. Because
of this potential ocular hazard, the procedure must be conducted in
a laser safe environment by a trained user.
Clinical Assessments
[0311] Patients were assessed by examiners who were unaware of the
treatment group. All examiners were trained on the NIHSS and
certified on the mRS. The NIHSS is a neurological function scale
that ranges from 0 to 42: scores between 7 and 22 are considered to
represent moderate to severe neurological impairment. The mRS is a
disability index ranging from 0 (no symptoms) to 6 (death). Outcome
measures (mRS and NIHSS) were assessed, in addition to baseline, at
5, 30, 60, and 90 days. Baseline data were collected including age,
sex, patient demographics, time from stroke onset to arrival at
hospital, time to treatment, prestrike mRS, vital signs, and a
complete medical history.
Safety Assessments
[0312] Vital signs, neurological scores, concomitant medications,
adverse events, and serious adverse events were recorded from study
entry to day 90. Unresolved serious adverse events were followed
for an additional 30 days.
[0313] The Data Monitoring Committee examined rates of death,
adverse events, and serious adverse events, as well as anticipated
and unanticipated device effects during the study. After evaluating
the safety data for the first 100 and 400 patients, they did not
stop further recruitment into the trial. Because of rapid
enrollment, there was no efficacy interim analysis. Neuroimaging
assessments were completed at baseline and at 5-day follow-up .+-.2
days.
Statistical Analysis
[0314] All analyses are intention-to-treat. Outcomes were obtained
from two 90-day scores, the mRS and the NIHSS score. Patients
without a day-90 visit had their last observation carried forward.
The mRS outcomes were: (1) the entire ordinal scale 0 to 6; and (2)
a dichotomous outcome with a favorable outcome (success) defined as
a 0 to 2 score and an unfavorable outcome (failure) defined as a 3
to 6 score. The NIHSS outcomes were: (1) the change in score from
baseline to 90 days using the entire ordinal scale 0 to 42 (with
death scored as 42); and (2) a dichotomous outcome for which
success could be achieved in 2 ways, either as a 90-day score of 0
to 1 or as a beneficial change from baseline to 90 days of 9 or
more points.
[0315] The primary efficacy outcome measure was the dichotomous mRS
90-day end point with success (mRS 0 to 2) and failure (mRS 3 to
6). The null hypothesis that the proportion of successes did not
differ by treatment, was tested using multiple logistic regression
with 2 prespecified covariates: (1) stroke severity at baseline;
and (2) time from stroke onset to time of randomization (TFSO).
Baseline stroke severity had 3 levels: NIHSS score 7 to 10
(moderate), 11 to 15 (moderately severe), and 16 to 22 (severe).
TFSO had 2 levels: 0 to 12 hours and 12 to 24 hours. The
randomization procedure was balanced on (but not stratified by)
these factors.
[0316] Secondary analyses determined the sensitivity of results to
the choice of covariates, of test procedure, and of analytic
outcome. The logistic model was run on the primary outcome measure
with additional covariates: age, sex, history of CAD, history of
diabetes, and history of stroke. To vary the test procedure of
logistic regression with a dichotomous outcome, we used the
nonparametric `shift` test with the same dichotomous outcome.
Explicitly, the shift test is the Cochran-Mantel-Haenszel (CMH)
test with ridit scores to rank outcomes (the van Elteren test,
Stokes M E, Davis C S, Koch G C. Categorical Data Analysis Using
the SAS System II Cary, N.C.: SAS Institute, Inc; 2000). In the
stroke literature, the shift test has often only been applied to
the full range of the mRS scale, but we applied it to all
dichotomous and ordinal outcomes. Logistic regression as run using
only dichotomous outcomes. All tests were adjusted for baseline
severity and TFSO. Odds ratios were calculated only for tests with
dichotomous outcomes.
[0317] The safety end points included mortality and all adverse
events including procedure and device related adverse events.
Safety end points were summarized with descriptive statistics and
tables. The analyses were carried out in SAS version 9.1.
Results
Baseline Characteristics
[0318] The patients were enrolled between January 2007 and April
2008. A total 660 patients were randomized (331 TLT and 327 sham),
as shown in FIG. 35. There were 7 (1.1%) patients lost to
follow-up. The groups were balanced with respect to baseline
characteristics (see Table 9). Mean time from stroke onset was
14.6.+-.5.9 hours (range 2.7 to 23.9) for the TLT group and
14.7.+-.6.1 hours (range 2.5 to 23.9) for the shame group; median
times were 15 hours and 16 hours for the TLT group and sham group,
respectively. Baseline NIHSS means scores were 13.1 (range 7 to 22)
for TLT and 13.2 (range 7 to 23) for shame. Median NIHSS scores
were 12 for TLT and 13 for sham.
TABLE-US-00009 TABLE 9 Demographic and Baseline Characteristics TLT
Sham (n = 331) (n = 327) Age (v) 70.4 .+-. 12.6 70.0 .+-. 11.9 Male
(%) 55.3 57.8 Ethnicity (No., %) White 256 (77%) 254 (77%) Black 37
(11%) 38 (12%) Hispanic 7 (2%) 8 (2%) Other (Largely Mestizo and
Native American 31 (10%) 29 (9%) Indians) Time Median time to
treatment (h) 15 16 Mean time to treatment (h:min) 14.38 .+-. 5:55
14:43 .+-. 6:12 Min (h:min) 2:42 2:30 Max (h:min) 23:54 23:54 Range
(h) 3-24 3-24 Time to treatment strata (%) <12 h 35.6 35.6 12-24
h 64.4 64.4 NIHSS score Mean 13.1 .+-. 4.7 13.2 .+-. 4.6 NIHSS
score strata (%) Median 12 13 Mean 7-10 39.0 38.9 11-15 26.6 27.1
16-22 34.4 34.0 History (%) Diabetes 31.4 33.4 Hypertension 82.2
83.9 Atrial fibrillation 38.7 35.6 Current smoker 20.2 19.1
Clinical Outcomes
[0319] Of the 660 patients: 331 received TLT and 327 received sham;
120 (36.3%) in the TLT group achieved successful outcome versus 101
(30.9%), in the sham group (P=0.094). OR 1.38 (95% CI, 0.95 to
2.00) as shown in FIG. 36, which shows the distribution of scores
on the mRS. The scores on the mRS indicate the following: 0, no
symptoms at all; 1, no significant disability despite symptoms; 2,
slight; 3, moderate; 4, moderately severe; 5, severe disability; 6,
death.
[0320] The results of secondary analyses on the prespecified
outcomes were consistent with those of the primary analysis. The
logistic regression analysis of the primary outcome measure with
additional covariates yielded a treatment OR-1.34 (95% CI, 0.94 to
2.03). All covariates except sex and history of stroke were
significant. With success defined as a mRS score of 0 to 2, the
shift test, stratified for the baseline severity and TFSO, showed a
nonsignficiant trend toward better outcomes with TLT having a
probability value of 0.091 and had an OR 1.38 (95% Cl, 0.95 to
2.01). For the ordinal mRS, scores 0 to 6, the shift test trended
in favor of TLT with a probability value of 0.113. The odds ratios
for all subsets on the primary outcome measure showed a
nonsignificant trend favoring TLT treatment except for those
patients with severe strokes at baseline. FIG. 37 shows the effects
of TLT on the primary end point, on subsets of the data defined by
categories of selected baseline characteristics. The simple
unadjusted odds ratios for TLT as compared to sham for the 90-day
outcome of success defined as mRS score 0 to 2. The horizontal
lines indicate 95% CIs. Also shown are the probability values for
the test of an interaction between the primary end point and the
categories within each subgroup. CAD indicates coronary artery
disease; TFSO, time from stroke onset to randomization.
[0321] For the dichotomous NIHSS score outcome for which success
had 2 components, the shift test trended in favor of TLT with an OR
of 1.23 (95% CI, 0.88 to 1.73) and a probability value of 0.23.
Logistic regression analysis results for the dichotomous NIHSS
outcome were very similar. An extension of the logistic regression
analysis separated the 2 components of success, a 90-day NIHSS
score of 0 to 1 and change from baseline .gtoreq.9 points, and
treated them as correlated outcomes: the OR was 1.30 (95% CI, 0.96
to 1.75) in favor of TLT with a probability value of 0.086. For the
ordinal outcome of change in NIHSS score from baseline to 90 days,
controlling for baseline severity and TFSO, the shift test
indicated the TLT group and had a probability value of 0.65. In
patients with deep infarcts, there was no difference in response
between the active and sham groups.
Safety Analysis
[0322] Mortality rates, serious adverse event (SAE) rates, and
adverse event (AE) rates were virtually the same. The TLT and sham
groups, respectively, had 58 (17.5%) and 57 (17.4%) deaths, 125
(37.8%) and 137 (41.8%) SAEs; and 92.7% and 93.6% of subjects,
respectively, had at least 1 adverse event. No SAEs were directly
attributable to TLT. The proportion of patients in the TLT group
showing hemorrhagic transformation at day 5 was 49 (14.8%) in the
TLT group and 56 (17.1%) in the sham group. There was no difference
in the safety outcomes between the 2 groups.
Discussion
[0323] The NEST-2 trial results presented here provide evidence of
the safety of TLT. The effective size of TLT for the treatment of
ischemic stroke in humans within 24 hours of stroke onset was
inadequate to meet conventional levels of statistical significance
for efficacy, even when the corrections were made for the baseline
imbalances in stroke severity and time to treatment, but showed a
consistent signal toward better outcomes associated with TLT.
[0324] Baseline severity was prespecified into 3 categories:
NIHSS=7 to 10, 11 to 15, and 16 to 22. Patients with baseline NIHSS
16 to 22 had a combined dichotomous mRS success rate of 8% (n=224;
TLT 7.0%, sham 9.1%). When restricted to the 434 patients with
moderate and moderately severe stroke (baseline NIHSS score 7 to
15), a post hoc analysis found a significant beneficial effect
(P=0.044). For these 434 patients, the dichotomous mRS success rate
of TLT showed an absolute improvement rate of 9.7% (TLT 51.6%, sham
41.9%). A similar beneficial effect was also found in the NEST-1
trial on the dichotomous mRS.
[0325] The failure to initially find the optimal treatment
population is reminiscent of the tPA development program. When tPA
therapy was administered between 0 and 6 hours, the treated groups
improved compared to the placebo ones on all measures, but the
final results did not achieve statistical significance. Again,
there was a strong signal of efficacy, but it was not until several
trials were completed and the proper inclusion and exclusion
criteria were established that the clinical benefit of tPA was
unequivocally established. TLT may take additional trial(s) to find
the treatment groups that are indisputably helped by the therapy.
In contrast to tPA, however, TLT has no untoward side effects, so
the barrier to treatment should be much lower.
[0326] There are potential weaknesses in the study that should be
noted. There was insufficient previous human experience with TLT to
be able to correctly power the study. The trial could have excluded
patients with severe strokes at baseline and a prestroke mRS
.gtoreq.2. Also, it is possible that TLT does not have nay effect
on stroke recovery. However, substantial preclinical studies, the
NEST-1 study, and the trends in the current trial argue against
this unfavorable interpretation.
[0327] A new feature of this form of therapy is that that
electromagnetic energy, in the past, has been used almost
exclusively for its destructive actions, such as burning out parts
of solid tumors with a gamma knife, various types of radiation
therapy, and both skin lesion removal and assorted opthalmologic
uses. Preclinical studies have demonstrated that infrared energy
produces potential beneficial action by alteration of biochemical
pathways, and that these changes are not due to thermal effects.
Based on preclinical studies, it is thought that when administering
TLT, the temperature of the brain is insignificantly elevated, and
the energy is producing its beneficial actions by alteration of
some biochemical reactions. It is known that infrared energy can
stimulate mitochondria, increase ATP formation, mitigate apoptosis,
and possibly enhance neurorecovery mechanisms. The precise nature
and balance of these reactions in the ischemic human brain are not
fully understood; therefore, the exact mechanism of action of TLT
remains unknown. Nevertheless, this docs open up a whole new range
of potential photobiology therapies for a variety of disorders.
Brain trauma and hemorrhagic stroke are obvious extensions; it is
also possible that TLT will be useful for a range of
neurodegenerative diseases that may involve mitochondrial
dysfunction.
Conclusion
[0328] The NEST-2 included a very broad range of stroke patients
with respect to baseline severity, prestrike disability, and time
to treatment. In this overall population, TLT was safe but did not
significantly improve patient outcomes as measured by both mRS and
NIHSS; however, both outcome measures showed positive trends in
that direction. Post hoc analyses suggest a meaningful beneficial
effect in patients with moderate to moderately severe ischemic
stroke within 24 hours of onset. Further clinical studies in this
redefined population should be considered.
Phototherapy Example 3
[0329] Another example examined the efficacy of low-level laser
therapy (LLLT) for TBI using the mouse closed-head injury (CHI)
model by studying the neurobehavioral and histological outcome of
the traumatized mice (A. Oron et al., "Low-Level Laser Therapy
Applied Transcranially to Mice following Traumatic Brain Injury
Significantly Reduces Long-Term Neurological Deficits," Journal of
Neurotrauma, Volume 24, Number 4, 2007 which is incorporated in its
entirety by reference herein.)
[0330] A total of 24 male Sabra mice (Hebrew University strain)
weighing 25-35 g were studied. The mice were housed in groups of
six per cage, under a 12 h/12 h, light/dark, reversed light cycle.
Food and water were provided ad libitum. The study was performed
according to the guidelines of the Institutional Animal Care
Committee in The Hebrew University, Jerusalem, Israel. CHI trauma
was induced to the heads of the mice under ether anesthesia using a
weight-drop device. Following sagittal scalp incision, mice were
manually immobilized, and a tipped Teflon cone was placed 3 mm
lateral to the midline and 1 mm caudal to the left coronal suture.
Following this, a metal rod (94 g) was allowed a free fall on the
cone from a precalibrated height.
[0331] Motor function and reflexes of the traumatized mice were
evaluated at different time intervals after CHI using a
neurological severity score (NSS), which was modified to emphasize
the motor functions. The neurological tests were based on the
ability of the mice to perform 10 different tasks (listed in Table
10) that evaluate the motor ability, balancing, and alertness of
the mouse. One point is given for failing to perform each of the
tasks: thus, a normal, uninjured mouse scores zero. The severity of
injury is defined by the initial NSS, evaluated one hour post-CHI
and referred to as NSS1, and is a reliable predictor of the late
outcome. Thus, fatal or near-fatal injury is defined in mice as
having a NSS1 of 9-10; severe injury in mice with an NSS1 of 7-8;
moderate injury with NSS1 of 5-6, and mild injury in mice with an
NSS1 of less than 4.
TABLE-US-00010 TABLE 10 Task NSS Presence of mono- or hemiparesis 1
Inability to walk on a 3-cm-wide beam 1 Inability to walk on a
2-cm-wide beam 1 Inability to walk on a 1-cm-wide beam 1 Inability
to balance on a 1-cm-wide beam 1 Inability to balance on a round
stick (0.5 cm wide 1 Failure to exit a 30-cm-diameter circle (for 2
min) 1 Inability to walk straight 1 Loss of startle behavior 1 Loss
of seeking behavior 1 Maximum total 10
[0332] All mice were subjected to CHI and assigned to control or
laser-treated groups as described below. Assessment of neurological
score (NSS) was performed one hour post-injury. The NSS scores of
the mice ranged from 4 to 6, indicating a mild-moderate trauma. The
mice were then divided into three groups of eight mice per group,
so that the means NSS in each group were similar, to ensure similar
average severity of injury in all groups. Two groups of mice
received low level light therapy at two different doses (10 and 20
mW/cm.sup.2) while the third group, serving as a sham-operated
control, underwent the same procedures as the laser-treated group,
but did not receive actual laser irradiation.
[0333] Laser treatment was performed four hours post-CHI using an
experimental laser 808-nm wavelength and maximal power output of
200 mW equipped with metal-backed glass fiber optic (3 mm
diameter). Laser light was applied by placing the distal tip of the
fiber optics in full contact with a point in the midline of the
skull (sagittal suture) located 4 mm caudal to the coronal suture
of the skull after skin had been removed in that region by a small
longitudinal incision. In preliminary experiments, the entire
cortical part of a fresh skull was excised from a mouse four hours
post-TBI. The distal tip of the fiber optic of the laser was placed
on the same location of the skull as described above. The power
density of the laser post-transmission through the skull was
measured by placing the probe of the laser power meter (Ophir Inc.,
Jerusalem, Israel) under the dura. Based upon prior measurements,
this location was chosen as being sufficient to illuminate the
entire brain (1.2-cm beam diameter measured by an infrared viewer)
due to dispersion of the laser beam by the skull. The laser
irradiation power density at the tip of the fiber optic was set to
deliver a power density of 10 and 20 mW/cm.sup.2 to the brain. The
duration of laser irradiation was two minutes (energy densities of
1.2-2.4 J/cm.sup.2). The power and energy densities of the laser
were chosen based on preliminary experiments with various
parameters of the laser applied in the CHI mouse model. Similar
laser parameters were used in previous rat and rabbit stroke models
where beneficial effects were evident.
[0334] Cortical tissue loss was evaluated in the traumatized
cross-section of the brain. At the end of the four-week follow-up
period, mice were anesthetized, and their brains were perfused with
4% paraformaldehyde solution and fixed in the same formaldehyde
solution for three days. Two-mm-thick cross-sections were taken
from the trauma region of the brain. This region was located in the
parietal part of the brain by means of a small blood clot that
served as a good marker of the spot where the skull was hit during
the induction of the trauma. The 2-mm-thick brain slices were
processed in an automated tissue processor and embedded in
paraffin. Eight-micron-thick coronal sections were prepared from
each 2-mm-thick brain slice block. Three random sections from each
brain slice were stained with hematoxylin and eosin, and processed
for quantitative measurements of the lesion area in each section.
The brain area was calculated in each hemisphere [ipsilateral
(traumatized) and contralateral] separately in each histological
section. The brain hemisphere volume was then calculated by
multiplying average area of the three sections by 2-mm thickness of
the slice. Lesion volume (in percentage) was expressed as
contralateral (nontraumatized hemisphere) volume minus ipsilateral
hemisphere volume divided by the contralateral hemisphere
volume.
[0335] Data were analyzed using one-way analysis of variance
(ANOVA) followed by Student Neuman Keuls test (using Sigma Stat
software from Sigma of St. Louis, Mo.). Data from this study
discussed below are presented as mean.+-.standard deviation. A
value of p less than 0.05 was regarded as statistically
significant.
[0336] The severity of injury as reflected by the NSS at one hour
was similar in both groups of mice: 4.8.+-.0.7 and 4.8.+-.0.8 in
the control (n=8) and laser-treated (n=16) mice, respectively.
Since during the whole follow-up period there was no statistical
difference between the mice that received 10 and 20 mW/cm.sup.2,
the results were pooled together to one laser-treated group of
mice.
[0337] FIG. 39 is a plot of the neurologic score (NSS) of
control/non-laser-treated (light columns) and laser-irradiated
(dark columns) mice at different time intervals after induction of
brain trauma. At 24 hours post-CHI, no significant differences
between sham-laser, control, traumatized mice, and those that had
received active laser treatment four hours post-trauma were
observed. Yet, from 48 hours on, there was a significant
improvement in the NSS in the laser-treated mice as compared to
control non-treated mice (p=0.05 at 48 hours and p<0.05 at later
times). The significant differences between the laser-treated and
control mice were sustained thereafter up to 28 days post-TBI,
where the neurobehavioral scores were 26-27% lower in the
laser-treated versus control mice.
[0338] FIG. 40 shows representative micrographs of cross-sections
of control non-laser-treated and laser-treated mice brains. Note
lesion (L) in the control micrograph as compared to minimal lesion
(arrow) in the laser-treated brain. FIG. 41 is a plot of lesion
volume of control (light column) and laser-treated (dark column)
traumatized brains 28 days after induction of head trauma (scale
bar=5 mm; **p<0.01). At 28 days post-TBI, the sham control mice
showed a significantly higher (p<0.001) loss (12.1.+-.1.3%) of
cortical tissue at the injured site compared to the laser-treated
mice (1.4.+-.0.1%).
[0339] The results of the study demonstrate for the first time that
low level laser therapy given at four hours post-TBI improves
short- and long-term (28 days) neurobehavioral function and reduces
brain tissue loss. The results corroborate previous findings in rat
and rabbit models that low-level light therapy applied
transcranially improves functional outcome after stroke. This
result is consistent with the hypothesis that pathophysiologies and
secondary mechanisms of damage of cerebral ischemia and head trauma
are quite similar, with a common involvement of both harmful and
protective mechanisms. In recent studies on the effect of low-level
light therapy on stroked rats, however, it was shown that the
beneficial effect on neurological function was evident only 2-3
weeks after laser application, whereas in the study described
herein, the marked improvement in overall neurological function of
the laser-treated mice over sham control mice was significant
already at five days post-trauma. This phenomenon can be partially
explained based on rapid elevation of ATP content after laser
irradiation in the ischemic heart. Furthermore, increase in total
antioxidants, angiogenesis, heat shock proteins content, and
antiapoptotic activity following low-level laser therapy were found
previously for the ischemic heart and skeletal muscles and can be
suggested as possible processes that are attenuated by the
low-level laser therapy also in the ischemic/traumatic brain. The
modulation of the above processes by low-level laser therapy could
contribute in concert to the better neurological outcome and the
significant reduction in lesion size of the laser-treated mice as
compared to controls as found in this study. Another mechanism that
may contribute to the long-term improvement of neurological
function in the laser-irradiated mice post-TBI include neurogenesis
that is induced by low-level laser therapy as found recently in
laser-treated stroked rats. Neurogenesis may also partially account
for the significant reduction in lesion size in the laser-treated
mice compared to control as demonstrated in this study.
[0340] The results of this study may also have clinical relevance.
The laser treatment was given four hours post-trauma, which is a
reasonable time for a patient to arrive at the hospital emergency
room. In a prior safety study in rats, no alterations in functional
neurological outcome or brain histopathology that had been treated
with low-level laser therapy at different intensities or
frequencies over both short- and long-term time intervals were
shown.
Phototherapy Example 4
[0341] Photobiostimulation effects of near-infrared light therapy
(NILT) have been documented for both in vitro and in vivo models
(see, e.g. L. DeTaboada et al., "Transcranial application of
low-energy laser irradiation improves neurological deficits in rats
following acute stroke," Lasers Surg. Med. 38:70-73 (2006)). Energy
in the infrared region of the electromagnetic spectrum is
non-ionizing and poses none of the hazards associated with
ultraviolet light. Irradiation of brain tissue with specific
infrared wavelengths (e.g. 808-810 nanometers) can penetrate the
brain (see, e.g. S. Ilic et al., "Effects of power densities,
continuous and pulse frequencies, and number of sessions of
low-level laser therapy on intact rat brain," Photomed. Laser Surg.
24:458-466 (2006)) and can stimulate adenosine triphiosphate (ATP)
formation by mitochondria (see, e.g. A. P. Sommer et al., "Stressed
cells survive better with light," J. Proteome. Res. 1:475 (2002)).
A primary mitochondrial chromophore responsible for
photobiostimulation is the enzyme cytochrome c oxidase, a terminal
enzyme in the cellular respiratory chain located in the inner
mitochondrial membrane. Cytochrome c oxidase (COX) plays a central
role in the bioenergetics of cells by delivering protons across the
inner membrane, thereby driving the formation of ATP by oxidative
phosphorylation. The overall result of NILT is improved energy
metabolism and potentially, enhanced cell viability (both neuronal
and non-neuronal).
[0342] NILT or photon energy delivered using a gallium-arsenic
diode near-infrared laser may be useful to treat the behavioral
function deficits associated with embolic stroke (see, e.g. P. A.
Lapchak et al., "Transcranial infrared laser therapy improves
clinical rating scores after embolic strokes in rabbits," Stroke
35:1985-1988 (2004)). These rabbit small clot embolic stroke model
(RSCEM) studies showed that NILT used in administered within 6
hours of an embolic stroke using a continuous wave (CW) of light at
high power densities (25 mW/cm.sup.2) and long treatment durations
(10 min.) significantly improved behavioral function. Lower
energies and shorter treatment durations improve behavior when
applied early on following embolization. However, there were also
documented unwanted side-effects related to using high power
densities, such as an increase in both skin and brain temperature
directly under the laser probe. Studies by Michael Chopp and
colleagues (see, e.g. A. Oron et al., "Low-level laser therapy
applied transcranially to rats after induction of stroke
significantly reduces long-term neurological deficits," Stroke
37:2620-2624 (2006)) using the rat middle cerebral artery occusion
(MCAO) model have also shown that laser therapy can significantly
improve behavioral deficits using a rodent stroke model. In
contrast to the RSCEM study cited above in which laser light
therapy was neuroprotective when applied within 6 hours
postembolization in the MCAO model, laser treatment was only
effective when applied 24 hours, but not 4 hours after stroke
onset. Laser light therapy in the MCAO model also attenuated
neurological deficits without a corresponding reduction in stroke
lesion volume. Since there was no apparent statistically
significant difference in the infarct area of control and
laser-irradiated rats with ischemic strokes, the authors
hypothesized that noninvasive laser light therapy intervention may
improve function by inducing neurogenesis. This was demonstrated
using immunocytochemistry for doublecortin, a marker of migrating
cells and bromodeoxyuridine, a marker of proliferating cells.
Nevertheless, in both models, laser light therapy using CW energy
delivery resulted in behavioral improvement.
[0343] CW NILT delivery and pulse wave (PW) frequency settings have
been examined to determine the most efficacious treatment regimen
using the RSCEM, which is produced by injection of blood clots into
the cerebral vasculature. The RSCEM, which is a model of multiple
infarct ischemia, results in ischemia-induced behavioral deficits
that can be measured quantitatively with a dichotomous clinical
rating scale. The use of clinical rating scores (a behavioral
endpoint) in rabbits parallels the use of the National Institutes
of Health Stroke Scale (NIHSS) in stroke patients, the primary
behavioral endpoint for all clinical trials of new treatment for
acute ischemic stroke. For the study described below, three
different treatment regimens were used: 1) CW power density of 7.5
mW/cm.sup.2; 2) pulse mode 1 (P1) using a frequency of 300 Us pulse
at 1 kHz; or 3) pulse mode 2 (P2) using a frequency of 2 ms pulse
at 100 Hz. The lower power density settings and pulse mode
frequencies have been shown to be safer methods of irradiation
because they produce little heating or no tissue damage.
[0344] Experimental Procedures
[0345] Surgery
[0346] The Department of Veterans Affairs approved the surgical and
treatment procedures used in this study. Surgery was done in a
sterile controlled environment with a room temperature between
22.8-23.2.degree. C. Using the RSCEM, male New Zealand White
rabbits weighing 2.2-2.5 kg were anesthetized with
halothane/O.sub.2 and a catheter was inserted into the common
carotid artery (CCA) through which microclots were injected.
Briefly, the bifurcation of one CCA was exposed and the external
carotid was ligated just distal to the bifurcation. A catheter was
inserted into the CCA and secured with ligatures. The incision was
closed around the catheter so that the distal end was accessible
outside the animal's neck. The catheter line was then filled with
heparinized saline and plugged with an injection cap. Rabbits were
allowed to recover from anesthesia until they were fully awake and
behaving normally. Once rabbits had recovered from anesthesia, they
were self-maintaining and have a normal core body temperature (see,
e.g. P. A. Lapchak et al., "Neuroprotective effects of the spin
trap agent
disodium-[(tert-butylimino)methyl]benzene-1,3-disulfonate N-oxide
(generic NXY-059) in a rabbit small clot embolic stroke model:
combination studies with the thrombolytic tissue plasminogen
activator," Stroke 33:1411-1415 (2002)).
[0347] Rabbit Embolic Strokes
[0348] For the RSCEM, blood was drawn from one or more donor
rabbits and allowed to clot for 3 hours at 37.degree. C. The large
blood clots were then suspended in phosphate-buffered saline (PBS)
containing 0.1% bovine serum albumin (BSA) and Polytron-generated
fragments were sequentially passed through metal screens and nylon
filters. The resulting small clot suspension was then labeled with
"cobalt containing NEN-Trac microspheres, so that the quantity of
clots that becomes lodged in the brain following embolization could
be calculated. For embolization, rabbits were placed in Plexiglas
restrainers, the catheter injection cap was removed and the
heparinized saline was cleared from the carotid catheter system.
Then, the clot particle suspension was rapidly injected through the
catheter into the brain, which was then flushed with sterile
saline. Rabbits were fully awake during the embolization procedure
and they were self-maintaining (i.e., they did not require
artificial respiration or other external support). The use of awake
non-anesthetized rabbits allows for immediate observation of the
effects of embolization on behavior at the time of clot injection
and thereafter.
[0349] Clinical Rating Score Behavioral and Quantal Dose--Response
Analysis
[0350] The use of clinical rating scores and quantal analysis is a
sophisticated method to determine how a large population of stroke
"patients" in this case, rabbits, will respond to a treatment and
is an appropriate primary endpoint to use when a treatment is being
developed to support a clinical trial. To evaluate the quantitative
relationship between clot dose and behavioral deficits, logistic
sigmoidal quantal analysis curves were fitted to the dose. A wide
range of clot doses was used to produce behaviorally normal or
abnormal animals. In the absence of a neuroprotective treatment
regimen, small number of microclots causes no grossly apparent
neurologic dysfunction. However, when large numbers of microclots
are injected, they invariably caused encephalopathy and sometimes
death. Abnormal rabbits with encephalopathy include those with one
or more of the following symptoms: ataxia, leaning, circling,
lethargy, nystagmus, loss of balance, loss of limb/facial sensation
and occasionally, paraplegia. Using a simple dichotomous rating
system, with a reproducible composite result and low inter-rater
variability (<5%), each animal was rated as either behaviorally
normal or abnormal by a naive observer. Using quantal analysis,
behavioral changes was detected following pharmacological
intervention. With this simple rating system, the composite result
for a group of animals is quite reproducible. A separate curve was
generated for each treatment condition and a statistically
significant increase in the value or the amount of microclots that
produce neurologic dysfunction in 50% of a group of animals
compared with control is indicative of a behavioral
improvement.
[0351] Power and Statistical Analysis
[0352] Power analysis of the quantal dose-response curves indicates
that, assuming .alpha.=0.05 and .beta.3=0.90, a coefficient of
variation of 15% and a difference between means of 20%, a sample
size of 14 animals is required per group. However, previous
experience shows that approximately 20 animals per group are needed
due to premature losses caused by preparation difficulties or death
after ischemia, but before treatment can be fully administered. The
behavioral data are presented as P.sub.50 (mean.+-.S.E.M.) in mg
clots for the number of rabbits in each group (n). P.sub.50 values
are analyzed using the t-test, which included the Bonferroni
correction where appropriate. In the study, a full sham-treated
control group was run parallel to the treatment groups and all
groups are rated using the same behavioral analysis scale described
above to remove any experimental or investigator bias. NILT-treated
groups are directly compared with control groups. For all animals
in this study, prior to carotid catheter placement, the fur on the
scalp covering the skull of all rabbits used in this study was
shaved to reveal pink skin. All shaved rabbits were randomly
allocated into treatment groups before the embolization procedure,
with concealment of the randomization until all postmortem analyses
were complete.
[0353] Near Infrared Light Therapy
[0354] Rabbits were placed in a Plexiglas restrainer for the
duration of the treatment. The probe was placed in direct contact
with the skin, since the fur over the head region was shaved prior
to juxtaposing the laser probe to the skin. An Acculaser low energy
laser fitted with a fiberoptic cable and laser probe (OZ Optics
Ltd., Berkeley, Calif.) measuring 2 centimeters in diameter was
used (wavelength of 808.+-.5 nanometers). Instrument design studies
showed that these specifications would allow for light penetration
of the rabbit skull and brain to a depth of approximately 2.5-3
centimeters and that the light beam would encompass the majority of
the brain if placed on the skin surface posterior to bregma on the
midline. Using the specifications described above, the average
energy delivered to the brain was 0.9-1.2 joules. Table 11 provides
details for the three different treatment regimens used in the
present studies: 1) CW; 2) P1 using a frequency of 300 .mu.s pulse
at 1 kHz; and 3) P2 using a frequency of 2 ms pulse at 100 Hz.
TABLE-US-00011 TABLE 11 NILT treatment regimens Power On Treatment
Time post- Treatment density Pulse time Duty duration embolization
regimen (mW/cm.sup.2) (Hz) (ms) cycle (min.) (hours) CW 7.5 NA NA
100% 2 6 and 12 PW mode 1 7.5 1000 0.3 30% 2 6 and 12 PW mode 2 7.5
100 2 20% 2 6
[0355] Results
[0356] Transcranial NILT Improves Clinical Rating Scores: 6-Hour
Treatment
[0357] In this study, 48 hours has been used as the time to measure
the behavioral endpoint in determining the relative effectiveness
of different treatment regimens. In the previous 2004 Lapchak study
cited above, the improvements in behavior that were present at 48
hours were also measurable up to 21 days following treatment. In
this study, a power density setting of 7.5 mW/cm.sup.2 and
treatment duration of 2 min. were used. In order to define
effective regimens of NILT, the three different modes: CW, P1 and
P2 described above were used. The main difference between the three
modes is the duty cycle and the overall time for brain
irradiation.
[0358] FIGS. 42A-42C show the results of the studies using a 6-hour
post-embolization time of treatment. The dark circles represent the
raw data for the control group and the triangles represent the raw
data for the laser-treated group. A normal animal for a specific
clot weight is represented by a symbol plotted at 0 on the y axis,
whereas an abnormal animal for a specific clot weight is
represented by a symbol plotted at 100 on the y axis. NILT
initiated 6 hours following embolization using all three treatment
regimens improved behavioral performance compared with a cumulative
control group when behavior was measured 24 hours following
treatment. As shown in FIG. 42A, NILT given as a CW increased the
P.sub.50 value to 2.06.+-.0.59 mg (n=29, P=0.099) and this was not
statistically different from the control group P.sub.50 value which
was 1.01.+-.0.25 mg (n=31). However, when NILT was given either as
a PW using the P1 or P2 modes, there were significant increases in
behavioral function that were reflected by significantly increased
P.sub.50 values for the groups. The P.sub.50 values were
1.89.+-.0.29 mg (n=25, P=0.0248) and 1.92.+-.0.15 mg (n=33,
P=0.0024), respectively for the P1 and P2 groups, as shown in FIGS.
42B and 42C.
[0359] Effect of Transcranial NILT on Behavioral Function when
Initiated 12 Hours Following Embolization
[0360] In the previous 2004 Lapchak study cited above, laser
therapy was reported to not be effective when applied to rabbits 24
hours following a stroke. Thus, it is important to determine if the
therapeutic window for NILT could be extended past 6 hours using
either the CW or P1 regimens. For this series of experiments, the
cumulative control group described above was used, a group that had
a P.sub.50 value of 1.01+0.25 mg (n=31). This cumulative control
group was used because it has been previously documented that the
baseline P.sub.50 value for embolized rabbits is quite consistent
over time and between experiments. The use of a cumulative control
group in this study allows the reduction of the utilization of
large number of rabbits.
[0361] FIGS. 43A and 43B show the results of the studies using a
12-hour post-embolization time of treatment. The dark circles
represent the raw data for the control group and the triangles
represent the raw data for the laser-treated group. A normal animal
for a specific clot weight is represented by a symbol plotted at 0
on the y axis, whereas an abnormal animal for a specific clot
weight is represented by a symbol plotted at 100 on the y axis.
NILT initiated 12 hours following embolization using either of the
treatment regimens did not significantly improve behavioral
performance compared with the cumulative control group when
behavior was measured 24 hours following treatment. As shown in
FIG. 43A, NILT given as a CW increased the P.sub.50 value to
2.89.+-.1.76 mg (n=29, P=0.279). In addition, as shown in FIG. 43B,
when NILT was given as a PW using the P1 the P.sub.50 value was
2.40.+-.0.99 mg (n=24, P=0.134). Thus, although there was an
increase in the mean P.sub.50 values, inherent variability in the
response of the treatment group to NILT prevented the treatment
from showing statistically significant differences. Since neither
the CW nor the P1 regimens were effective at significantly
increasing the P.sub.50 value, the effects of NILT using the P2
regimen at the 12-hour post-embolization time point were not
investigated.
[0362] Discussion
[0363] The principal aim of this study was to directly evaluate the
effects of CW and PW transcranial NILT on clinical rating scores
following multiple small clot embolic strokes in rabbits. To
achieve this goal, the RSCEM was used which is a useful preclinical
animal model to develop new stroke treatments that can be quickly
translated to the clinic. In the model, the primary cause of stroke
or ischemic injury is related to the administration of blood clots
that lodge in cerebral vessels, the same cause of many strokes in
humans responsible for the interruption of cerebral blood flow and
clinical deficits. Moreover, RSCEM is a model of multiple infarct
ischemia that uses a scale of clinical rating scores as the primary
endpoint which is based upon motor function components of the NIHSS
for stroke in humans.
[0364] In the present study that used the RSCEM, transcranial NILT
applied to embolized rabbits 6 hours following an embolic stroke
improved behavioral rating scores, specifically motor function.
There are three important aspects of these findings. First, the low
power density (7.5 mW/cm.sup.2) with an estimated brain penetration
of 2.5-3 cm as used in the study was effective at improving
clinical rating scores (e.g. motor function). Second, a 2 minute
application of near infrared light promoted recovery of function
when NILT is applied transcranially to embolized rabbits. Third,
the studies were designed to determine if there were differential
effects of CW therapy compared with pulse therapy. Interestingly,
although the CW regimen increased the P.sub.50 value of group, the
resulting increase was not statistically significant from control.
However, both pulse treatment regimens significantly increased the
P.sub.50 value of group compared with control.
[0365] In the previous 2004 Lapchak study, laser therapy at a high
power density (25 mW/cm.sup.2) was tested using a CW regimen for a
10 minute duration to show that laser therapy could improve
behavioral function following embolic strokes in rabbits. The
present study shows that the power density of 7.5 mW/cm.sup.2 and
treatment time of 2 minutes can produce beneficial effects at the
extended 6 hour post-embolization treatment time. However, in this
study, CW therapy did not result in "statistically" significant
behavioral improvements. But pulse therapy, whether using the P1 or
P2 regimen, did result in a statistically significant increase in
behavior. These results with low power density NILT can be compared
with those previously reported using a rodent stroke model which
had some significant differences with the present study. Using a
rodent filament model to produce permanent cerebral ischemia, the
rat MCAO study cited above was unable to show a benefit to early
laser treatment (i.e., 4 hours following filament placement).
However, this study found that laser treatment was effective when
applied 24 hours following the stroke and that the beneficial
effect was observable up to 21 days following the stroke. The
durability of the laser therapy effects is comparable to that
previously published. However, even though the previous 2004
Lapchak study indicated that NILT attenuates stroke-induced
behavioral deficits in rabbits when applied up to 6 hours following
the stroke, it did not show any "statistically" significant
behavioral improvement when NILT was started 12 hours following the
stroke.
[0366] There appears to be an important difference between the
results published for the rodent filament permanent occlusion model
cited above and the rabbit embolic stroke model 2004 study of
Lapchak cited above. The difference may simply be associated with
the cause of stroke in the two models. The placement of a plastic
filament into the middle cerebral artery of a rodent that is being
maintained on 1-2% halothane, 70% N.sub.2O and 30% O.sub.2 and
being monitored for blood glucose, pH, pO.sub.2 and pCO.sub.2 and
rectal temperature (all levels were undefined in the publication)
may represent an extreme artificial model of cerebral stroke. The
rabbit embolic stroke model as described within used rabbits 3
hours following the placement of a carotid catheter to ensure that
the rabbits were self-maintaining and free of anesthetic when they
were embolized. Moreover, fresh blood clots prepared from donor
rabbits were used for the embolization procedure on the day that
the blood was drawn and allowed to clot free of additives. Thus,
during the embolization procedure, rabbits were awake and behaving
normally. With such differences between the two models,
differential effects of NILT are not too surprising. The different
findings resulting from the use of two diverse animal models of
ischemic stroke may shed some light on the mechanism(s) of action
of NILT in the ischemic brain. In the rat model study, it was
proposed that laser therapy may be inducing neurogenesis in the
brain primarily because the effect of laser was only evident when
applied 24 hours following ischemia and that there was coincident
elevation of neurogenesis-associated markers in the brain.
Moreover, the physiological effects of laser therapy were not
observed immediately, but required a delay of between 2 and 4 weeks
post-treatment. The authors suggested that laser therapy induces
the formation of new neurons and neuron-supporting cells in the
brain and that the behavioral improvement following laser therapy
requires the new neuronal pathways. Unfortunately, this idea was
not supported by measures of hematoxylin and eosin (H&E)
staining for lesion volume in the laser-treated and control
ischemic brains as there was no statistically significant
difference in lesion volume between the two groups, suggesting that
there was little or no neurogenesis in brain.
[0367] In contrast to the delayed effects of laser therapy in the
rat model, in the rabbit model, NILT is effective when applied 5
minutes to 6 hours following embolization and the behavioral
effects of NILT are measurable within 24-48 hours of embolization.
The results described herein suggest that NILT is inducing a rapid
response element (RRE) in brain following embolization that results
in behavioral improvement, an effect that appears to be independent
of neurogenesis or long-term remodeling. Moreover, in stroke
patients entered in the NeuroThera Effectiveness and Safety Trial
(NEST-1), a significant reduction in NIHSS score was evident by day
5 after laser treatment, which continued to improve up to 90 days
after treatment (see, Y. Lampl et al., "Infrared laser therapy for
ischemic stroke: a new treatment strategy: results of the
NeuroThera Effectiveness and Safety Trial-1 (NEST-1)," Stroke
38(6): 1843-1849 (2007)). This finding is in agreement with the
results of the rabbit embolic stroke model studies, which show that
laser therapy induces rapid recovery of function following an
ischemic stroke.
[0368] Taken together, results from experimental stroke studies
suggest that NILT may be useful in patients if applied between 6
and 24 hours of a stroke. Based upon correlative preclinical and
clinical acute ischemic stroke studies, the 6 hour therapeutic
window observed in the RSCEM may be equivalent to approximately 18
hours in a stroke patient because of the following. In the RSCEM,
tPA effectively improves behavior when given 1-1.5 hours following
embolization and in patients the therapeutic window for tPA is
along the order of 3-6 hours after stroke onset. Thus, there
appears to be a two- to threefold differential between the observed
times for effectiveness of tPA in the rabbit model compared with
patients. Consistent with this hypothesis, in the RSCEM, NILT was
effective up to 6 hours following a stroke and as predicted by the
model, the NEST-1 clinical trial cited above found that laser
therapy improved clinical rating scores if patients were treated
within 24 hours following a stroke. Thus, because of the time
differential discussed above, it would appear that NILT may be
useful in patients if they present at a clinic within 24 hours of a
stroke.
[0369] The NEST-1 study cited above has provided some indication
that transcranial laser therapy may be a useful approach to
treating motor function deficits resulting from ischemic strokes
when laser therapy is initiated 2-24 hours following the stroke.
The first trial of the NEST-1 study used a CW mode with a power
density of 10 mW/cm.sup.2 at 20 pre-determined locations on the
scalp for 2 minutes of irradiation at each site. The preliminary
results from the NEST-1 trial of 120 Acute ischemic stroke patients
(biased design of 1:2) with a treatment time between 2 and 24 hours
(median time 18 hours) showed that more patients in the
laser-treated group had successful outcomes from baseline to 90
days, measured by a mean change in the NIHSS and mRS scores, than
did controls (P=0.035, stratified by severity and time to
treatment; P=0.048, stratified by severity alone). The positive
results documented for NILT in animal models of stroke prompted the
initiation of the NEST-II trial, a prospective, intent-to-treat,
multi-center, international, double-blinded trial sponsored by the
manufacturers of the transcranial laser device, PhotoThera Inc.
[0370] NILT effectively improves behavioral deficits when applied
to the rabbit brain using PWs of light energy. While the data do
support the use of NILT therapy in a clinical setting, the
differential results obtained using CW and PW treatment suggest
that the use of a PW mode of NILT will be most beneficial to
patients to improve motor function and overall well-being.
Phototherapy Example 5
[0371] In another example study performed under contract with
Neurological Testing Service, Inc. (Charleston, S.C.), Infrared
Transcranial Laser Therapy (TLT) was tested for efficacy in an
amyloid precursor peptide (APP) transgenic mouse model of
Alzheimer's Disease (AD). Laser light therapy was administered
three times per week at various doses for 26 weeks, starting at 3
months of age, and the results were compared to no laser (control
group). Animals were examined for amyloid load, inflammatory
markers, brain A.beta. levels, plasma A.beta. levels, CSF A levels,
sAPP levels, and NS behavioral changes. The number of A.beta.
plaques was significantly reduced in the brain with administration
of laser therapy in a dose dependent fashion. Administration of
laser therapy demonstrated a dose dependent reduction in amyloid
load. All therapies were effective in the reduction in amyloid
deposition. All laser treatments reduced the behavioral effects
seen with advanced amyloid deposition. Overall, laser therapy was
effective at limiting the extent of A.beta. amyloid in the brain
and reversing the effect of deposition of A.beta. peptide and
behavioral deficits in the mouse. In addition, the laser therapies
were able to reduce the expression of inflammatory markers in the
APP transgenic mice. A.beta. peptide levels were significantly
changed in the brains of the APP transgenic mice, and there was no
detectable difference in plasma A.beta. peptide levels. Studies
showed an increase in sAPP.beta. and a decrease in sAPP.beta.
levels consistent with inhibition of the .beta.-secretase activity.
These studies suggest that laser light therapy is a potential
candidate for therapeutic treatment of AD.
[0372] As used herein, the following abbreviations have the
following definitions:
TABLE-US-00012 Abbreviations Definition A.beta. Abeta APP Amyloid
Precursor Protein A.beta. peptide Abeta peptide AD Alzheimer's
Disease 4G8 Abeta peptide antibody PTI PhotoThera, Inc. NTS
Neurological Testing Service, Inc. BACE Beta secretase IL-1
Interleukin-1 TNF-.alpha. Tumor necrosis factor-.alpha. TGF-.beta.
Transforming growth factor-.beta.
[0373] A.beta. containing senile plaques are one of the
neuropathological hallmarks of Alzheimer's Disease (AD) and a
considerable effort has been expended in understanding the
relationship of A.beta. and A.beta.-containing senile plaques to
AD. Much of this work has focused on the biosynthesis of A.beta.
and factors that influence its deposition. The A.beta. peptides are
primarily two peptides of either 40 or 42 amino acids generated via
internal proteolysis of its precursor, the amyloid precursor
protein (APP). In addition to A.beta.-containing senile plaques, a
variety of neuronal cytoskeletal alterations are prominent features
of AD neuropathology. These include phospho-tau containing
neurofibrillary tangles, free-lying dystrophic neurites and those
present in neuritic senile plaques, and synapse loss. Whether these
abnormal features are the result of or cause of neuronal loss is
still controversial. Regardless of the precise mechanism, this
neuronal and synaptic loss leads to cognitive decline. Early onset
autosomal dominant AD is directly linked to mutations in one of
several genes: APP, presenilin 1 (PS1), or presenilin 2 (PS2). In
addition several risk factor genes, most notably the APOE4 allele,
alter risk for later onset AD, and it is clear that mutations or
polymorphisms in several other genes can lead to similar AD
phenotypes.
[0374] The A.beta. peptide is derived from APP, which is cleaved by
the sequential action of the .beta.- and .gamma.-secretases. The
.beta.-site APP cleavage enzyme (BACE) is a member of the membrane
bound aspartyl proteases which results in the cleavage of APP on
the extracellular side of the membrane releasing the soluble
APP-.beta. (sAPP.beta.) fragment. In addition, the
.gamma.-secretase enzyme (a complex of PS-1 and PS-2) cleaves the
transmembrane domain to release the A.beta. peptide and carboxyl
terminus. The .alpha.-secretase enzyme is the predominent APP
activity that cleaves in the middle of the A.beta. peptide and
prevents the generation of the A.beta. peptide. Altered functions
of these enzymes can lead to the enhanced production of A.beta.
peptide, which may contribute to AD pathogenesis. A number of
studies have shown that mutations in the APP gene or in presenilins
result in the increase in. .beta.-secretase cleavage and the
production of both A.beta.1-40 and A.beta.1-42. In addition, a
depletion of cholesterol using cholesterol lowering-agents produced
a decrease in A.beta. peptide synthesis and sAPP-. Therefore,
understanding the mechanisms associated with altered A.beta.
processing and the role of .beta.-secretase in the process will
help in the design of selective inhibitors of .beta.-secretase and
eventually therapeutic treatment of AD.
Methods and Materials
[0375] The amyloid precursor protein (APP) transgenic model of
mouse A.beta. peptide amyloidosis was used. APP transgenic mice
were administered no laser or laser therapy as outlined below
3.times./week for 26 weeks starting at 3 months of age. At the end
of the experiment, animals were subject to behavioral analysis,
were sacrificed and the brains were divided in half and prepared as
follows: 1/2 brain was examined for A.beta. plaque burden in the
brain (i.e., plaque number), and inflammatory markers and the
second 1/2 of the brain was homogenized for brain A.beta. peptide
level and sAPP levels. Animals were treated daily at 1 pm and were
tested on days 176-179 for the behavioral studies and the final
trials were performed on the 26.sup.th week (four hours after the
treatment). Animals were sacrificed immediately after training and
plasma, CSF and brain were collected for analysis.
[0376] A control APP group was used to determine the baseline of
amyloid deposits (treatment was simulated with the laser disabled,
no laser energy). The group started as 3 month old mice and
maintained in the study for 26 weeks to reach 9 months of age. In
addition, at the end of the study, the animals were subjected to
behavioral (Morris water maze) analysis. NTS was not blinded to the
study parameters. The laser was prepared by PTI and shipped to NTS.
The animals were subjected to behavioral studies, amyloid load,
A.beta. peptide analysis, inflammatory markers, sAPP levels, brain
and plasma for A.beta. analysis. The animals in each of the groups
were allowed to complete the study and all protocols were carried
out after 26 weeks. Endpoints were as follows: [0377] Amyloid load
in brain (left hemisphere) [0378] Inflammatory markers in brain
(IL-1, TNF-alpha, TGF-.beta.) [0379] A.beta. 1-40, 1-42 in brain
(1/2 from animals/plasma from all animals) [0380] Plasma A.beta.
levels (13 and 26 weeks: 4 hrs post dose). [0381] sAPP.alpha. and
.beta. levels from brain. Brain collection (week 26 only) for
brain/plasma/CSF A.beta. peptide levels. [0382] Gross necropsy
examination [0383] Behavior analysis--MWM performed during the last
week of treatment, animals to be sacrificed after last
administration. Test includes tracking of swimming distance and
time to reach platform.
[0384] Male APP transgenic mice (NTS, Inc.) weighing approximately
35-40 grams each were given free access to food and water before
and during the experiment. The animals were administered laser
therapy. The laser was prepared by PTI and delivered to NTS. The
APP mice (male) used in this experiment were designed by
microinjection of the human APP gene (with the Swedish and London
mutations) into mouse eggs under the control of the
platelet-derived growth factor B (PDGF-B) chain gene promoter. The
mice were generated on a C57BL/6 background and were developed by
MTI. Animals were housed in the Medical University of South
Carolina Animal Facility under a 12:12 light:dark cycle. Animals
were housed in standard non-sterile rodent microisolator cage, with
filtered cage top and housed 4 to a cage. Animals were fed ad
libitum and maintained by brother sister mating. Transgenic animals
were identified by PCR analysis. The mice generated from this
construct, develop amyloid deposits starting at 6 months of age.
Animals were aged for 3 months and then maintained for 26 weeks and
sacrificed for amyloid quantification.
[0385] For histological examination, the animals were anesthetized
with an intraperitoneal injection of sodium pentobarbital (50
mg/kg). The animals were transcardially perfused with 4.degree. C.,
phosphate-buffered saline (PBS) followed by 4% paraformaldehyde.
The brains were removed and placed in 4% paraformaldehyde over
night. The brains were processed to paraffin and embedded. Ten
serial 30-.mu.m thick sections through the brain were obtained.
Tissue sections were deparaffinized and washed in Tris buffered
saline (TBS) pH 7.4 and blocked in the appropriate serum (mouse).
Sections were blocked overnight at 4.degree. C. and then subjected
to primary antibody overnight at 4.degree. C. (A.beta. peptide
antibody, 4G8, Signet) in order to detect the amyloid deposits in
the brain of the transgenic animals. Sections were washed in TBS
and secondary antibody (Vector Laboratories) was added and
incubated for 1 hour at room temperature. After washing the
sections were incubated as instructed in the Vector ABC Elite kit
(Vector Laboratories) and stained with diaminobenzoic acid (DAB).
The reactions were stopped in water and cover slipped after
treatment to xylene. The amyloid area in each section was
determined with a computer-assisted image analysis system,
consisting of a Power Macintosh computer equipped with a Quick
Capture frame grabber card, Hitachi CCD camera mounted on an
Olympus microscope and camera stand. NIH Image Analysis Software,
v. 1.55 was used. The images were captured and the total area of
amyloid was determined over the ten sections. A single operator
blinded to treatment status performed all measurements. Summing the
amyloid volumes of the sections and dividing by the total number of
sections calculated the amyloid volume per animal.
[0386] For quantitative analysis, we used an enzyme-linked
immunosorbent assay (ELISA) to measure the levels of human
A.beta..sub.1-40 and A.beta..sub.1-42 in the brains of APP
transgenic mice (IBL, 27718 for 1-40 and 27711 for 1-42).
A.beta..sub.1-40 and A.beta..sub.1-42 were extracted from mouse
brains as described below: [0387] 1. Weighed frozen hemi-brains.
[0388] 2. Prepared Tissue Homogenization Buffer (THB-see following
recipe) by adding Protease Inhibitor Cocktail (PIC, Sigma) 1:1000
dilution immediately before use. [0389] 3. Homogenized hemi-brains
in 1 mL of THB+PIC per each 100 mg of tissue (e.g. 2.2 mL of buffer
was used to homogenize a hemi-brain weighing 220 mg). [0390] 4.
Homogenized brains (using polytron) and samples were aliquoted and
snap frozen in liquid nitrogen.
Tissue Homogenization Buffer (THB):
[0390] [0391] (250 mM sucrose, 20 mM tris base, 1 mM EDTA, 1 mM
EGTA)
TABLE-US-00013 [0391] 5 mL 1M Tris base (pH 7.4) 21.4 g sucrose 0.5
mL 0.5 M EDTA 1.0 mL 0.25 EGTA
[0392] Added ddH.sub.20 to 250 mL. [0393] Sterile filtered and
handled aseptically. [0394] Stored at 4.degree. C.
[0395] ELISA assays were performed as described by the manufacturer
(IBL): [0396] 1) The wells for reagent blank were determined. 100
.mu.L each of "4, EIA buffer" was put into the wells. [0397] 2)
Wells for test sample blank, test sample and diluted standard were
determined. Then, 100 .mu.L each of test sample blank, test sample
and dilutions of standard were placed into the appropriate wells.
[0398] 3) The precoated plate for were incubated overnight at
4.degree. C. after covering it with plate lid. [0399] 4) Each well
of the precoated plate was washed vigorously with wash buffer using
washing bottle (3 times). Then, each well was filled with wash
buffer and place the precoated plate for 15.about.30 seconds. Wash
buffer was removed completely from the precoated plate by snapping.
This procedure was repeated more than 7 times. Then, the remaining
liquid was removed from all wells completely by snapping the
precoated plate onto paper towel. [0400] 5) 100 .mu.L of labeled
antibody solution was pipetted into the wells of test samples,
diluted standard and test sample blank. [0401] 6) The precoated
plate was incubated for 1 hour at 4.degree. C. after covering it
with plate lid. [0402] 7) The precoated plate was washed 9 times in
the same manner above (4). [0403] 8) The required quantity of "6,
Chromogen" was pipetted into a disposable test tube. And then, 100
.mu.L was pipetted from the test tube into the wells. [0404] 9) The
precoated plate was incubated for 30 minutes at room temperature in
the dark. [0405] 10) 100 .mu.L of "7, Stop solution" was pipetted
into the wells. The liquid was mixed by tapping the side of
precoated plate. [0406] 11) The plate reader was run and
measurement conducted at 450 nm. The measurement shall be done
within 30 minutes after the addition of "7, stop solution".
[0407] For quantitative analysis of A.beta. peptide levels in the
plasma and CSF, an enzyme-linked immunosorbent assay (ELISA) was
used to measure the levels of human A.beta..sub.1-40 and
A.beta..sub.1-42 in the plasma and CSF of APP transgenic mice (IBL,
27718 for 1-40 and 27711 for 1-42). A.beta..sub.1-40 and
A.beta..sub.1-42 ELISAs were performed as above. Blood was collect
by saphenous vein collection or cardiac puncture (terminal bleed)
in lithium:heparin and plasma was prepared by centrifugation. CSF
was collected an analyzed.
[0408] For sAPP assays in the hemibrain, Western blot analysis,
extracts containing 20-50 .mu.g of total protein were mixed with
Tris/glycine reducing buffer, denaturing loading buffer, loaded,
and electrophoresed on 8% Tris/glycine gels (Invitrogen). Gels were
transferred to nitrocellulose membranes, incubated with the
respective primary antibodies followed by secondary antibodies
conjugated to horseradish peroxidase and processed for
visualization by enhanced chemiluminescence ECL Plus.TM. (Amersham
Biosciences). The 6E10 monoclonal Ab (Signet, Dedham, Mass.)
recognizes the first 17 amino acids of the A.beta. peptide. 6E10 Ab
was used for Western blotting to detect soluble APP.alpha.. Rabbit
869 antibody was used to detect sAPP.beta. by Western blotting.
Secondary antibodies conjugated to horseradish peroxidase were from
Jackson ImmunoResearch (West Grove, Pa.).
[0409] For immunohistochemical analysis for cytokine evaluation in
the hemibrain, tissue sections were deparaffinized and washed in
Tris buffered saline (TBS) pH 7.4 and blocked in the appropriate
serum (goat). Sections were blocked overnight at 4.degree. C. and
then subjected to primary antibody overnight at 4.degree. C.
Sections were washed in TBS and secondary antibody was added and
incubated for 1 hour at room temperature. After washing the
sections were incubated as instructed in the Vector ABC Elite kit
and stained with diaminobenzoic acid (DAB). The reactions were
stopped in water and cover slipped after treatment to xylene.
[0410] Morris water-maze testing was used for behavioral analysis.
All mice were tested once in the Morris water maze test at the end
of the experiment. Mice were trained in a 1.2 m open field water
maze. The pool was filled to a depth of 30 cm with water and
maintained at 25.degree. C. The escape platform (10 cm square) was
placed 1 cm below the surface of the water. During the trials, the
platform was removed from the pool. The cued test was carried out
in the pool surrounded with white curtains to hide any extra-maze
cues. All animals underwent non-spatial pretraining (NSP) for three
consecutive days. These trials are to prepare the animals for the
final behavioral test to determine the retention of memory to find
the platform. These trials were not recorded (for training purposes
only). For the training and learning studies, the curtains were
removed to extra maze cues (this allowed for identification of
animals with swimming impairments). On day 1, the mice were placed
on the hidden platform for 20 seconds (trial 1), for trials 2-3
animals were released in the water at a distance of 10 cm from the
cued-platform or hidden platform (trial 4) and allowed to swim to
the platform. On the second day of trials, the hidden platform was
moved randomly between the center of the pool or the center of each
quadrant. The animals were released into the pool, randomly facing
the wall and were allowed 60 seconds to reach the platform (3
trials). In the third trial, animals were given three trials, two
with a hidden platform and one with a cued platform. Two days
following the NSP, animals were subjected to final behavioral
trials (Morris water maze test). For these trials (3 per animal),
the platform was placed in the center of one quadrant of the pool
and the animals released facing the wall in a random fashion. The
animal was allowed to find the platform or swim for 60 seconds
(latency period, the time it takes to find the platform). All
animals were tested within 4-6 hours of dosing and were randomly
selected for testing by an operator blinded to the test group.
Animals were tested on days 176-179 for the non-spatial pretraining
and the final trials were performed on day 180.
[0411] The results are expressed as the mean.+-.standard error of
mean (SEM). The significance of differences in the amyloid and
behavioral studies were analyzed using a t-test. Comparisons were
made between the 9-month-old APP control group (baseline
group--started at 3 months of age) and the 9-month old treated
mice. Differences below 0.05 were considered significant. Percent
changes in amyloid and behavior were determined by taking the
summation of the data in each group and dividing by the comparison
(i.e., treated/9 month control=% change). Animals that developed
severe complications following administration of laser were
excluded from the study.
[0412] Animals (100 mice) were subjected to administration of no
laser or laser for two minutes 3.times./week beginning at 3 months
of age and continued for 6 months. Animals were male and were
randomly assigned to the different treatment groups per Table 12
and Table 13.
TABLE-US-00014 TABLE 12 Radiant Beam Irradiance Regime Temporal
Power at diameter at at the (Treatment Treatment Mode.sup.(1)
Format the Skin.sup.(2) the Skin Skin.sup.(3) Frequency) Regime
Control (20) OFF 0 mW N/A 0 W/cm.sup.2 3 .times. Week CW (20)
Continuous 40 mW 3 mm 1.3 W/cm.sup.2 3 .times. Week Pulsed I (20)
ON = 2 mSec; 40 mW 3 mm 1.3 W/cm.sup.2 3 .times. Week @ 100 Hz
Pulsed II (20) ON = 2 mSec; 200 mW 3 mm 6.4 W/cm.sup.2 3 .times.
Week @ 100 Hz Pulsed III (20) ON = 2 mSec; 400 mW 3 mm 12.7
W/cm.sup.2 3 .times. Week @ 100 Hz .sup.(1)Total Number of Animals
= 100 .sup.(2)These values are based on tissue transmission and
scattering measurements done on mice at earlier TLT studies
(Ischemic Stroke and TBI), and updated based on measurements of
scalp/skull transmission and scatter on two APP mice. The measured
data was used to calculate the Radiant Power at the skin - Table
13, needed to deliver the irradiances listed to the animals Dura.
.sup.(3)These are calculated values showing average Irradiances for
Continuous TLT and Peak Irradiances for Pulsed TLT. Irradiance at
the Skin = Radiant Power/Area of beam = Radiant Power/(.pi. * (0.1
cm).sup.2) = Radiant Power/0.0314 cm.sup.2
TABLE-US-00015 TABLE 13 Group Name for Radiant Tables and Power at
Irradiance at Treatment Mode Figures Temporal Format the Skin the
Dura.sup.(1) Regime Control A OFF 0 mW 0 mW/cm.sup.2 CW B
Continuous 40 mW 10 mW/cm.sup.2 Pulsed I C ON = 2 mSec; @ 100 Hz 40
mW 10 mW/cm.sup.2 Pulsed II D ON = 2 mSec; @ 100 Hz 200 mW 50
mW/cm.sup.2 Pulsed III E ON = 2 mSec; @ 100 Hz 400 mW 100
mW/cm.sup.2 (1) These are calculated values showing average
Irradiances for Continuous TLT and Peak Irradiances for Pulsed
TTL.
Results
[0413] The laser was provided as a powder to NTS. Amyloid load was
determined in the animals treated with laser therapy and no laser.
Table 14 and FIG. 44 illustrate the results. The no laser group
demonstrated a .about.2% amyloid burden which is the standard level
of amyloid in this particular model at .about.9 months of age
(previous studies). The laser therapy demonstrated a dose dependent
attenuation of the amyloid load when compared to the vehicle group.
At all doses except CW, the amount of amyloid actually was lower
than in the 9 month control group indicating that the laser therapy
not only stopped amyloid deposition, but may have even reversed the
level of amyloid. This suggests that the laser therapy was capable
of attenuating the amyloid in these mice. There were no deaths in
this study. Animals were examined for gross abnormalities following
sacrifice. No gross pathological features were detected in the
animals.
TABLE-US-00016 TABLE 14 Percent decrease in amyloid in the brain
Percent change A.beta. amyloid Burden Treatment in A.beta. amyloid*
% +/- SEM P-value** No laser NA 1.741 .+-. 0.1120 NA CW -11.8%
1.535 .+-. 0.04399 0.0951 Pulse I -54% 0.8015 .+-. 0.05021
<0.0001 Pulse II -67% 0.5740 .+-. 0.03374 <0.0001 Pulse III
-37.3% 1.091 .+-. 0.09192 <0.0001 *Percent changes are compared
to no laser **P value compared to no laser
[0414] The behavioral effects (behavior and distance) of treatment
with laser therapy were determined in the APP transgenic mice at
the termination of the experiment. Mice were subjected to the
Morris water maze task and the latency period and distance were
determined. Table 15 and FIG. 45A illustrate the results. The no
laser control demonstrated a latency time of 48.58 seconds, which
is the standard in this particular model at 9 months of age
(previous studies). All the laser treated animals demonstrated a
significant difference in latency when compared to the control.
This suggests that laser therapy was capable of attenuating the
latency time in these mice.
TABLE-US-00017 TABLE 15 Behavioral changes in APP transgenic mice
treated with LLT. % Change in Treatment Latency Period* Latency
time (sec) P value** No laser NA 48.58 .+-. 1.421 NA CW -34.5%
31.82 .+-. 1.382 <0.0001 Pulse I -50.5% 24.05 .+-. 1.060
<0.0001 Pulse II -68.8% 15.15 .+-. 1.082 <0.0001 Pulse III
-48.6% 24.99 .+-. 0.9763 <0.0001 *Percent changes are compared
to no laser **P value compared to no laser
[0415] Table 16 and FIG. 45B illustrates the results comparing
distance traveled in the water maze. The vehicle control
demonstrated a distance of 75.53 in, which is the standard in this
particular model at .about.9 months of age (previous studies). All
the animals demonstrated a significant difference in distance when
compared to the control group. This suggests that laser therapy was
capable of attenuating the behavioral effects in these mice.
TABLE-US-00018 TABLE 16 Behavioral changes in APP transgenic mice
treated with LLT. % Change in Treatment Distance* Distance (in) P
value** No laser NA 75.53 .+-. 2.746 NA CW -22.9% 58.27 .+-. 2.710
<0.0001 Pulse I -36.1% 48.24 .+-. 3.184 <0.0001 Pulse II
-51.5% 36.60 .+-. 2.737 <0.0001 Pulse III -29.6% 53.15 .+-.
2.206 <0.0001 *Percent changes are compared to no laser **P
value compared to no laser
[0416] The effect of laser therapy was determined on the expression
of inflammatory markers (IFMs) in the brain of APP transgenic mice
(FIGS. 46A-46C and Table 17). Table 17 and FIGS. 46A-46C illustrate
the results of the animals terminated at 26 weeks after the start
of treatment. The no laser control demonstrated specific staining
for inflammatory markers (IL-1, TNF and TGF-.beta.) as indicated in
Table 17, which are the standard in this particular model at
.about.9 months of age (previous studies). Laser therapy at all the
doses demonstrated a significant difference from the control
animals. This suggests that laser therapy was capable of
attenuating the IFMs in these mice.
TABLE-US-00019 TABLE 17 Changes in inflammatory markers in APP
transgenic mice treated with LLT*. % Change in IL-1 % Change in TNF
% Change in TGF-.beta. Treatment intensity units +/- SEM) intensity
units +/- SEM) intensity units +/- SEM) No laser 0% (112.6 .+-.
4.305) 0% (178.0 .+-. 10.26) 0% (137.6 .+-. 5.428) CW -15.3% (95.35
.+-. 2.164) -16.9% (148.0 .+-. 5.494) -11.9% (121.2 .+-. 2.597) p =
0.010 p = 0.0139 p = 0.0096 Pulse I -23.6% (85.80 .+-. 2.448)
-35.3% (115.2 .+-. 6.161) -24.6% (103.7 .+-. 3.373) p = >0.0001
p = >0.0001 p < 0.0001 Pulse II -48.9% (57.50 .+-. 2.948)
-57.5% (75.68 .+-. 6.875) -36.8% (86.99 .+-. 3.660) p = >0.0001
p = >0.0001 p < 0.0001 Pulse III -36.1% (71.92 .+-. 2.512)
-40.8% (105.3 .+-. 9.080) -24.4% (104.0 .+-. 3.836) p = >0.0001
p = >0.0001 p < 0.0001 *Percent changes are compared to no
laser
[0417] The effect of laser therapy was determined on the changes in
A.beta. peptide in the brain of APP transgenic mice (FIGS. 47A and
47B and Tables 18 and 19). Measurement of A.beta.1-40 and
A.beta.1-42 peptide (guanidine extractable) in the brain showed an
increase as the APP transgenic mice aged from 3 to 9 months. Table
18 and FIG. 47A illustrates the results of the animals terminated
at 26 weeks after the start of treatment. The control demonstrated
the levels of A.beta.1-40 in the brain as indicated in Table 18,
which is the standard in this particular model at .about.9 months
of age (previous studies). Laser therapy at all doses demonstrated
a decrease in A.beta.1-40 when compared to the control and they
were significant. This suggests that laser therapy was capable of
attenuating the A.beta.1-40 increase in these mice.
TABLE-US-00020 TABLE 18 Changes in A.beta. peptide levels in the
brain of APP transgenic mice % Change in A.beta.1-40 peptide levels
Treatment (pg/mg tissue +/- SEM)* P value** No laser NA (1849 .+-.
117.4) NA CW -37% (1165 .+-. 68.12) <0.0001 Pulse I -57.1%
(792.6 .+-. 68.39) <0.0001 Pulse II -74.3% (476.2 .+-. 59.75)
<0.0001 Pulse III -62% (702.2 .+-. 50.10) <0.0001 *Percent
changes are compared to no laser **P value compared to no laser
[0418] Table 19 and FIG. 47B illustrates the results of the animals
terminated on the 26.sup.th week after the start of treatment for
A.beta.1-42 peptide levels. The control demonstrated the levels of
A.beta.1-42 in the brain as indicated in Table 19, which is the
standard in this particular model at .about.9 months of age
(previous studies). Laser therapy at all doses demonstrated a
significant decrease in A.beta.1-42 when compared to control. This
suggests that laser therapy was capable of attenuating the
A.beta.1-42 peptide levels in these mice.
TABLE-US-00021 TABLE 19 Changes in A.beta. peptide levels in the
brain of APP transgenic mice % Change in A.beta.1-42 peptide levels
Treatment (pg/mg tissue +/- SEM) P value** No laser NA (1306 .+-.
86.60) NA CW -31.4% (896.4 .+-. 31.95) <0.0001 Pulse I -62%
(495.7 .+-. 67.13) <0.0001 Pulse II -76.4% (308.2 .+-. 44.68)
<0.0001 Pulse III -63.9% (471.3 .+-. 64.11) <0.0001 *Percent
changes are compared to no laser **P value compared to no laser
[0419] The effect of laser therapy was determined on the changes in
A.beta. peptide levels in the plasma of APP transgenic mice (FIGS.
48A and 48B and Tables 20 and 21). Measurement of total A.beta.
peptide in the plasma was performed on weeks 13 and 26 in the APP
transgenic mice treated from 3 to 9 months. As seen in the Tables
20 and 21 and FIGS. 48A and 48B, the total plasma A.beta. peptide
levels were similar in all groups for the 90 day time period. As
the study progressed and the animals were treated with laser
therapy, there was a significant change in the apparent plasma
total A.beta. peptide profile. This suggests that laser therapy was
capable of lowering the plasma A.beta. peptide levels, while brain
A.beta. peptide levels also changed.
TABLE-US-00022 TABLE 20 Changes in plasma total A.beta. peptide
levels in the APP transgenic mice treated with LLT. Week 13. %
Change in total A.beta. peptide levels Treatment (pg/ml plasma +/-
SEM) P value** No laser NA (17.52 .+-. 0.08758) NA CW +101.5%
(17.78 .+-. 0.1051) 0.0580 Pulse I -3.8% (16.86 .+-. 0.09818)
<0.0001 Pulse II -1.7% (17.23 .+-. 0.1104) 0.0469 Pulse III
-4.9% (16.67 .+-. 0.1206) <0.0001 *Percent changes are compared
to no laser **P value compared to no laser
TABLE-US-00023 TABLE 21 Changes in plasma total A.beta. peptide
levels in the APP transgenic mice treated with LLT. Week 26. %
Change in total A.beta. peptide levels Treatment (pg/ml plasma +/-
SEM) P value** No laser NA (187.8 .+-. 7.337) NA CW -21.7% (147.0
.+-. 6.521) <0.0001 Pulse I -35.3% (121.6 .+-. 6.325) <0.0001
Pulse II -39.8% (113.0 .+-. 5.840) <0.0001 Pulse III -17.7%
(154.5 .+-. 7.647) <0.0001 *Percent changes are compared to no
laser **P value compared to no laser
[0420] The effects of treatment with TTP-5854 laser therapy on
sAPP.alpha. and CTF.beta. levels in the brain were determined in
the APP transgenic mice at the termination of the experiment. Brain
tissue was subjected to extraction and Western blot analysis for
sAPP.alpha. and CTF.beta. levels following treatment. Tables 22 and
23 illustrate the results of the animals terminated at 26 weeks
after the start of treatment. The control demonstrated a given
level of sAPP.alpha. and CTF.beta. in the brain, which is the
standard in this particular model at .about.9 months of age
(previous studies). Laser therapy demonstrated a significant
difference in both sAPP.alpha. and CTF.beta. levels when compared
to the control. For sAPP.alpha., laser therapy at all doses
demonstrated significant differences from the control (Table 22 and
FIG. 49A). For CTF.beta., again all doses of laser therapy
demonstrated a significant difference from the control (Table and
FIG. 49B). This suggests that laser therapy was capable of
increasing the sAPP.alpha. and decreasing CTF.beta. in these mice
suggesting a shift from .beta.- to .alpha.-secretase activity.
TABLE-US-00024 TABLE 22 Changes in sAPP.alpha. levels in the brain
of APP transgenic mice % Change in sAPP.alpha. levels*(band
intensity Treatment arbitrary units) +/- SEM?) P value** No laser
NA (76.74 .+-. 2.301) NA CW +108.2% (83.05 .+-. 2.104) 0.0463 Pulse
I +131.2% (100.7 .+-. 1.994) <0.0001 Pulse II +154.7% (118.7
.+-. 2.475) <0.0001 Pulse III +123.5% (94.80 .+-. 2.018)
<0.0001 *Percent changes are compared to no laser **P value
compared to no laser
TABLE-US-00025 TABLE 23 Changes in CTF.beta. levels in the brain of
APP transgenic mice % Change in CTF.beta. levels* (band intensity
arbitrary Treatment units +/- SEM) P value** No laser NA (123.4
.+-. 2.261) NA CW -4.8% (117.5 .+-. 2.142) 0.0645 Pulse I -19.5%
(99.31 .+-. 3.245) <0.0001 Pulse II -39.7% (74.45 .+-. 3.492)
<0.0001 Pulse III -15% (104.9 .+-. 3.147) <0.0001 *Percent
changes are compared to no laser **P value compared to no laser
[0421] The treatment with laser therapy on CSF A.beta. peptide
levels was determined in the APP transgenic mice at the termination
of the experiment (FIG. 50 and Table 24). Mice were subjected to
ELISA for total A.beta. peptide levels were determined because of
the small amount of CSF obtained. Table 24 illustrates the results
of the animals terminated at 26 weeks after the start of treatment.
The control demonstrated A.beta. peptide levels shown in Table 24,
which is the standard in this particular model at .about.9 months
of age (previous studies). CW and Pulse I demonstrated no
significant difference in CSF when compared to the control.
However, laser therapy at Pulse II and Pulse III demonstrated
significant differences from the control (Table 24 and FIG. 50).
This suggests that laser therapy was capable of attenuating the
A.beta. peptide levels in the brain in these mice at specific
doses.
TABLE-US-00026 TABLE 24 Changes in A.beta. peptide levels in the
CSF of APP transgenic mice % Change in CSF total A.beta. peptide
levels* (CSF total A.beta. peptide levels Treatment and units +/-
SEM) P value** No laser NA (237.8 .+-. 20.70) NA CW -9.8% (214.8
.+-. 30.64) 0.5407 Pulse I -20.7% (188.7 .+-. 12.99) 0.0598 Pulse
II -39.6% (141.6 .+-. 12.40) 0.0009 Pulse III -27.5% (172.4 .+-.
13.15) 0.0157 *Percent changes are compared to no laser **P value
compared to no laser
[0422] Statistical analysis was performed on the samples as
described by GraphPad Prism (Version 4.00). The changes in the
different parameters were significant to the 0.05 level or greater.
This is due not only to the number of animals in each group, but
also to the number of samples taken for each measurement from each
animal (20). Therefore, there is confidence in the numbers. The
numbers fall within the range of acceptable results.
[0423] Animals were examined for gross abnormalities following
sacrifice. No gross pathological features were detected in most of
the animals except for Group E (Pulse III). Animals showed some
lesioning in the brain following treatment. Animals after a few
weeks began to develop lesions on the skin. They were switched to a
regime of 1 minute treat, ice, 1 minute treat without further
injury.
Discussion
[0424] Transgenic mice expressing the mutant form of the human APP
gene begin to deposit amyloid fibrils by 6 months of age. This
process is associated with increased A.beta. peptide in the
deposits in the brain. Recent studies have shown that
administration of laser light therapy can be protective against
various neuronal injuries. In this study, LLT was tested in the APP
model to determine the efficacy on amyloid load, inflammatory
markers, brain A.beta. levels, plasma and CSF A.beta. levels, and
behavioral changes. The number of A.beta. plaques was significantly
reduced in the brain with administration of LLT in a dose dependent
fashion. Administration of LLT demonstrated variable effects and
with increasing doses showing the greatest effect of reduction of
amyloid deposition.
[0425] NTS was not blinded to the study as outlined by PTI.
Overall, laser light therapy was effective at limiting the extent
of A.beta. amyloid in the brain and altering the amount of
deposition of A.beta. peptide and behavioral deficits in the
mouse.
[0426] The explanations and illustrations presented herein are
intended to acquaint others skilled in the art with the invention,
its principles, and its practical application. Those skilled in the
art may adapt and apply the invention in its numerous forms, as may
be best suited to the requirements of a particular use.
Accordingly, the specific embodiments of the present invention as
set forth are not intended as being exhaustive or limiting of the
invention.
* * * * *