U.S. patent application number 11/110000 was filed with the patent office on 2005-11-10 for electromagnetic treatment apparatus and method for angiogensis modulation of living tissues and cells.
Invention is credited to DiMino, Andre', Pilla, Arthur A..
Application Number | 20050251229 11/110000 |
Document ID | / |
Family ID | 35196696 |
Filed Date | 2005-11-10 |
United States Patent
Application |
20050251229 |
Kind Code |
A1 |
Pilla, Arthur A. ; et
al. |
November 10, 2005 |
Electromagnetic treatment apparatus and method for angiogensis
modulation of living tissues and cells
Abstract
An apparatus and method for electromagnetic treatment of living
tissues and cells comprising: configuring at least one waveform
according to a mathematical model having at least one waveform
parameter, said at least one waveform to be coupled to a
angiogenesis and neovascularization target pathway structure;
choosing a value of said at least one waveform parameter so that
said at least waveform is configured to be detectable in said
angiogenesis and neovascularization target pathway structure above
background activity in said target pathway structure; generating an
electromagnetic signal from said configured at least one waveform;
and coupling said electromagnetic signal to said angiogenesis and
neovascularization target pathway structure using a coupling
device.
Inventors: |
Pilla, Arthur A.; (Oakland,
NJ) ; DiMino, Andre'; (Woodcliff Lake, NJ) |
Correspondence
Address: |
LEN TAYLOR, PATENT ATTORNEY
261 DAVENPORT STREET
SOMERVILLE
NJ
08876
US
|
Family ID: |
35196696 |
Appl. No.: |
11/110000 |
Filed: |
April 19, 2005 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60563104 |
Apr 19, 2004 |
|
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Current U.S.
Class: |
607/86 |
Current CPC
Class: |
C12M 35/02 20130101;
A61N 2/02 20130101; C12N 13/00 20130101; A61N 1/40 20130101 |
Class at
Publication: |
607/086 |
International
Class: |
A61H 021/00 |
Claims
What is claimed is:
1) A method for electromagnetic treatment of living tissues and
cells by enhancing angiogenesis and neovascularization comprising
the steps of: Configuring at least one waveform according to a
mathematical model having at least one waveform parameter, said at
least one waveform to be coupled to a angiogenesis and
neovascularization target pathway structure; Choosing a value of
said at least one waveform parameter so that said at least waveform
is configured to be detectable in said angiogenesis and
neovascularization target pathway structure above background
activity in said angiogenesis and neovascularization target pathway
is structure; Generating an electromagnetic signal from said
configured at least one waveform; and Coupling said electromagnetic
signal to said angiogenesis and neovascularization target pathway
structure using a coupling device.
2) The method of claim 1, wherein said at least one waveform
parameter includes at least one of a frequency component parameter
that configures said at least one waveform to repeat between about
0.01 Hz and about 100 MHz, a burst amplitude envelope parameter
that follows a mathematically defined amplitude function, a burst
width parameter that varies at each repetition according to a
mathematically defined width function, a peak induced electric
field parameter varying between about 1 .mu.V/cm and about 100
mV/cm in said target pathway structure according to a
mathematically defined function, and a peak induced magnetic
electric field parameter varying between about 1 .mu.T and about
0.1 T in said target pathway structure according to a
mathematically defined function.
3) The method of claim 1, wherein said angiogenesis and
neovascularization target pathway structure includes at least one
of ions and ligands.
4) The method of claim 1, further comprising the step of binding
ions and ligands to regulatory molecules in living cells and
tissues thereby modulating angiogenesis and neovascularization.
5) The method of claim 4, wherein said binding of ions and ligands
includes modulating Calcium to Calmodulin binding.
6) The method of claim 4, wherein said binding of ions and ligands
includes modulating growth factor production in living cells and
tissues.
7) The method of claim 4, wherein said binding of ions and ligands,
includes modulating cytokine production in living cells and
tissues.
8) The method of claim 4, wherein said binding of ions and ligands
includes modulating growth factors and cytokines relevant to
angiogenesis and neovascularization.
9) The method of claim 4, wherein said binding of ions and ligands
includes modulating angiogenesis and neovascularization for
treatment of bone fractures and disorders.
10) The method of claim 4, wherein said binding of ions and ligands
includes modulating angiogenesis and neovascularization for
treatment of cardiovascular diseases.
11) The method of claim 4, wherein said binding of ions and ligands
includes modulating angiogenesis and neovascularization for
treatment of cerebral diseases.
12) The method of claim 4, wherein said binding of ions and ligands
includes modulating angiogenesis and neovascularization for
treatment of cerebrovascular disease.
13) The method of claim 4, wherein said binding of ions and ligands
includes modulating angiogenesis and neovascularization for
treatment peripheral vascular disease.
14) The method of claim 4, wherein said binding of ions and ligands
includes modulating angiogenesis and neovascularization for
treatment of diseased or ischemic cells and tissues.
15) The method of claim 4, wherein said binding of ions and ligands
includes modulating angiogenesis and neovascularization for
treatment of an acute or chronic soft tissue wound.
16) The method of claim 4, wherein said binding of ions and ligands
includes modulating angiogenesis and neovascularization for
treatment of sprains strains and contusions.
17) An electromagnetic treatment apparatus for plants, animals, and
humans to enhance angiogenesis and neovascularization comprising: A
waveform configuration means for configuring at least one waveform
to be coupled to a angiogenesis and neovascularization target
pathway structure according to a mathematical model having at least
one waveform parameter capable of being chosen so that said at
least one waveform is configured to be detectable in said
angiogenesis and neovascularization target structure above
background activity in said angiogenesis and neovascularization
target pathway structure; An electromagnetic signal generating
means connected to said waveform device by at least one connecting
means for generating an electromagnetic signal from said configured
at least one waveform; and A coupling device connected by at least
one connecting means to said electromagnetic signal generating
device for coupling said electromagnetic signal to said
angiogenesis and neovascularization target pathway structure.
18) The electromagnetic treatment apparatus of claim 17, wherein
said at least one waveform parameter includes at least one of a
frequency component parameter that configures said at least one
waveform to repeat between about 0.01 Hz an about 100 MHz according
to a mathematical function, a burst amplitude envelope parameter
that follows a mathematically defined amplitude function, a burst
width parameter that varies at each repetition according to a
mathematically defined width function, a peak induced electric
field parameter varying between about 1 .mu.V/cm and about 100
mV/cm in said angiogenesis and neovascularization target pathway
structure according to a mathematically defined function, and a
peak induced magnetic electric field parameter varying between
about 1 .mu.T and about 0.1 T in said angiogenesis and
neovascularization target pathway structure according to a
mathematically defined function.
19) The electromagnetic signal generating means of claim 17 wherein
the signal is inductively coupled to living cells and tissues
wherein Calcium binding to Calmodulin is modulated.
20 ) The electromagnetic signal generating means of claim 17
wherein the signal is capacitively coupled to living cells and
tissues wherein Calcium binding to Calmodulin is modulated.
21) The electromagnetic signal generating means of claim 17 wherein
the signal is inductively coupled to living cells and tissues
wherein growth factors and cytokines relevant to angiogenesis and
neovascularization are modulated.
22) The electromagnetic signal generating means of claim 21 wherein
the growth factors include at least one of fibroblast growth
factors, platelet derived growth factors and interleukin growth
factors.
23) The electromagnetic signal generating means of claim 17 wherein
the signal is capacitively coupled to living cells and tissues
wherein growth factors and cytokines relevant to angiogenesis and
neovascularization are modulated.
24) The electromagnetic signal generating means of claim 23 wherein
the growth factors include at least one of fibroblast growth
factors, platelet derived growth factors and interleukin growth
factors.
25) The electromagnetic signal generating means of claim 17 wherein
the signal is inductively coupled to living cells and tissues to
modulate growth factor production.
26) The electromagnetic signal generating means of claim 17 wherein
the signal is capacitively coupled to living cells and tissues to
modulate growth factor production.
27) The electromagnetic signal generating means of claim 17 wherein
the signal is inductively coupled to living cells and tissues to
modulate cytokine production.
28) The electromagnetic signal generating means of claim 17 wherein
the signal is capacitively coupled to living cells and tissues to
modulate cytokine production.
29) The electromagnetic signal generating means of claim 17 wherein
the signal is inductively coupled to living cells and tissues to
modulate angiogenesis and neovascularization for the treatment of
bone fractures and disorders.
30) The electromagnetic signal generating means of claim 17 wherein
the signal is capacitively coupled to living cells and tissues to
modulate angiogenesis and neovascularization for the treatment of
bone fractures and disorders.
31) The electromagnetic signal generating means of claim 17 wherein
the signal is inductively coupled to living cells and tissues to
modulate angiogenesis and neovascularization for the treatment of
cardiovascular diseases.
32) The electromagnetic signal generating means of claim 17 wherein
the signal is capacitively coupled to living cells and tissues to
modulate angiogenesis and neovascularization for the treatment of
cardiovascular diseases.
33) The electromagnetic signal generating means of claim 17 wherein
the signal is inductively coupled to living cells and tissues to
modulate angiogenesis and neovascularization for the treatment of
cerebral diseases.
34) The electromagnetic signal generating means of claim 17 wherein
the signal is capacitively coupled to living cells and tissues to
modulate angiogenesis and neovascularization for the treatment of
cerebral diseases.
35) The electromagnetic signal generating means of claim 17 wherein
the signal is inductively coupled to living cells and tissues to
modulate angiogenesis and neovascularization for the treatment of
cerebrovascular disease.
36) The electromagnetic signal generating means of claim 17 wherein
the signal is capacitively coupled to living cells and tissues to
modulate angiogenesis and neovascularization for the treatment of
cerebrovascular disease.
37) The electromagnetic signal generating means of claim 17 wherein
the signal is inductively coupled to living cells and tissues to
modulate angiogenesis and neovascularization for the treatment of
peripheral vascular disease.
38) The electromagnetic signal generating means of claim 17 wherein
the signal is capacitively coupled to living cells and tissues to
modulate angiogenesis and neovascularization for the treatment of
peripheral vascular disease.
39) The electromagnetic signal generating means of claim 17 wherein
the signal is inductively coupled to living cells and tissues to
modulate angiogenesis and neovascularization for the treatment of
diseased or ischemic cells and tissues.
40) The electromagnetic signal generating means of claim 17 wherein
the signal is capacitively coupled to living cells and tissues to
modulate angiogenesis and neovascularization for the treatment of
diseased or ischemic cells and tissues.
41) The electromagnetic signal generating means of claim 17 wherein
the signal is inductively coupled to living cells and tissues to
modulate angiogenesis and neovascularization for the treatment of
an acute or chronic soft tissue wound.
42) The electromagnetic signal generating means of claim 17 wherein
the signal is capacitively coupled to living cells and tissues to
modulate angiogenesis and neovascularization for the treatment of
an acute or chronic soft tissue wound.
43) The electromagnetic signal generating means of claim 17 wherein
the signal is inductively coupled to living cells and tissues to
modulate angiogenesis and neovascularization for the treatment of
sprains strains and contusions.
44) The electromagnetic signal generating means of claim 17 wherein
the signal is capacitively coupled to living cells and tissues to
modulate angiogenesis and neovascularization for the treatment of
sprains strains and contusions.
Description
[0001] This application claims the benefit of U.S. Provisional
Application 60/563,104 filed Apr. 19, 2004.
BACKGROUND OF THE INVENTION
[0002] 1. Field of the Invention
[0003] This invention pertains generally to an apparatus and a
method for treatment of living tissues and cells by altering their
interaction with their electromagnetic environment. This invention
also relates to a method of modification of cellular and tissue
growth, repair, maintenance, and general behavior by application of
encoded electromagnetic information. More particularly this
invention relates to the application of surgically non-invasive
coupling of highly specific electromagnetic signal patterns to any
number of body parts. In particular, an embodiment according to the
present invention pertains to using pulsing electromagnetic fields
("PEMF") to enhance living tissue growth and repair via
angiogenesis and neovascularization by affecting the precursors to
growth factors and other cytokines, such as ion/ligand binding such
as calcium binding to calmodoulin.
[0004] 2. Discussion of Related Art
[0005] It is now well established that application of weak
non-thermal electromagnetic fields ("EMF") can result in
physiologically meaningful in vivo and in vitro bioeffects.
[0006] EMF has been used in applications of bone repair and bone
healing. Waveforms comprising low frequency components and low
power are currently used in orthopedic clinics. Origins of using
bone repair signals began by considering that an electrical pathway
may constitute a means through which bone can adaptively respond to
EMF signals. A linear physicochemical approach employing an
electrochemical model of a cell membrane predicted a range of EMF
waveform patterns for which bioeffects might be expected. Since a
cell membrane was a likely EMF target, it became necessary to find
a range of waveform parameters for which an induced electric field
could couple electrochemically at the cellular surface, such as
voltage-dependent kinetics. Extension of this linear model also
involved Lorentz force analysis.
[0007] A pulsed radio frequency ("PRE") signal derived from a 27.12
MHz continuous sine wave used for deep tissue healing is known in
the prior art of diathermy. A pulsed successor of the diathermy
signal was originally reported as an electromagnetic field capable
of eliciting a non-thermal biological effect in the treatment of
infections. PRF therapeutic applications have been reported for
reduction of post-traumatic and post-operative pain and edema in
soft tissues, wound healing, burn treatment and nerve regeneration.
Application of EMF for the resolution of traumatic edema has become
increasingly used in recent years. Results to date using PRF in
animal and clinical studies suggest that edema may be measurably
reduced from such electromagnetic stimulus.
[0008] Prior art considerations of EMF dosimetry have not taken
into account dielectric properties off tissue structure as opposed
to the properties of isolated cells.
[0009] In recent years, clinical use of non-invasive PRF at radio
frequencies comprised using pulsed bursts of a 27.12 MHz sinusoidal
wave, wherein each pulse burst comprises a width of sixty-five
microseconds, having approximately 1,700 sinusoidal cycles per
burst, and various burst repetition rates. By use of a
substantially single voltage amplitude envelope with each PRF
burst, one was limiting frequency components that could couple to
relevant dielectric pathways in cells and tissue.
[0010] Time-varying electromagnetic fields, comprising rectangular
waveforms such as pulsing electromagnetic fields, and sinusoidal
waveforms such as pulsed radio frequency fields ranging from
several Hertz to an about 15 to an about 40 MHz range, are
clinically beneficial when used as an adjunctive therapy for a
variety of musculoskeletal injuries and conditions.
[0011] Beginning in the 1960's, development of modern therapeutic
and prophylactic devices was stimulated by clinical problems
associated with non-union and delayed union bone fractures. Early
work showed that an electrical pathway can be a means through which
bone adaptively responds to mechanical input. Early therapeutic
devices used implanted and semi-invasive electrodes delivering
direct current ("DC") to a fracture site. Non-invasive technologies
were subsequently developed using electrical and electromagnetic
fields. These modalities were originally created to provide a
non-invasive "no-touch" means of inducing an electrical/mechanical
waveform at a cell/tissue level. Clinical applications of these
technologies in orthopaedics have led to approved applications by
regulatory bodies worldwide for treatment of fractures such as
non-unions and fresh fracture, as well as spine fusion. Presently
several EMF devices constitute the standard armamentarium of
orthopaedic clinical practice for treatment of difficult to heal
fractures. The success rate for these devices has been very high.
The database for this indication is large enough to enable its
recommended use as a safe, non-surgical, non-invasive alternative
to a first bone graft. Additional clinical indications for these
technologies have been reported in double blind studies for
treatment of avascular necrosis, tendinitis, osteoarthritis, wound
repair, blood circulation and pain from arthritis as well as other
musculoskeletal injuries.
[0012] Cellular studies have addressed effects of weak low
frequency electromagnetic fields, on both signal transduction
pathways and growth factor synthesis. It can be shown that EMF
stimulates secretion of growth factors after a short, trigger-like
duration. Ion/ligand binding processes at a cell membrane are
generally considered an initial EMF target pathway structure. The
clinical relevance to treatments for example of bone repair, is
upregulation such as modulation, of growth factor production as
part of normal molecular regulation of bone repair. Cellular level
studies have shown effects on calcium ion transport, cell
proliferation, Insulin Growth Factor ("IGF-II") release, and IGF-II
receptor expression in osteoblasts. Effects on Insulin Growth
Factor-I ("IGF-I") and IGF-II have also been demonstrated in rat
fracture callus. Stimulation of transforming growth factor beta
("TGF-.beta.") messenger RNA ("mRNA") with PEMF in a bone induction
model in a rat has been shown. Studies have also demonstrated
upregulation of TGF-.beta. mRNA by PEMF in human osteoblast-like
cell line designated MG-63, wherein there were increases in
TGF-.beta.1, collagen, and osteocalcin synthesis. PEMF stimulated
an increase in TGF-.beta.1 in both hypertrophic and atrophic cells
from human non-union tissue. Further studies demonstrated an
increase in both TGF-.beta.1 mRNA and protein in osteoblast
cultures resulting from a direct effect of EMF on a
calcium/calmodulin-dependent pathway. Cartilage cell studies have
shown similar increases in TGF-.beta.1 mRNA and protein synthesis
from EMF, demonstrating a therapeutic application to joint repair.
Various studies conclude that upregulation of growth factor
production may be a common denominator in the tissue level
mechanisms underlying electromagnetic stimulation. When using
specific inhibitors, EMF can act through a calmodulin-dependent
pathway. It has been previously reported that specific PEMF and PRF
signals, as well as weak static magnetic fields, modulate Ca.sup.2+
binding to CaM in a cell-free enzyme preparation. Additionally,
upregulation of mRNA for BMP2 and BMP4 with PEMF in osteoblast
cultures and upregulation of TGF-.beta.1 in bone and cartilage with
PEMF have been demonstrated.
[0013] However, prior art in this field does not configure
waveforms based upon a ion/ligand binding transduction pathway.
Prior art waveforms are inefficient since prior art waveforms apply
unnecessarily high amplitude and power to living tissues and cells,
require unnecessarily long treatment time, and cannot be generated
by a portable device.
[0014] Therefore, a need exists for an apparatus and a method that
more effectively modulate angiogenesis and other biochemical
processes that regulate tissue growth and repair, shortens
treatment times, and incorporates miniaturized circuitry and light
weight applicators thus allowing the apparatus to be portable and
if desired disposable. A further need exists for an apparatus and
method that more effectively modulates angiogenesis and other
biochemical processes that regulate tissue growth and repair,
shortens treatment times, and incorporates miniaturized circuitry
and light weight applicators that can be constructed to be
implantable.
SUMMARY OF THE INVENTION
[0015] An apparatus an a method for electromagnetic treatment of
living tissues and cells by altering their interaction with their
electromagnetic environment.
[0016] According to an embodiment of the present invention, by
treating a selectable body region with a flux path comprising a
succession of EMF pulses having a minimum width characteristic of
at least about 0.01 microseconds in a pulse burst envelope having
between about 1 and about 100,000 pulses per burst, in which a
voltage amplitude envelope of said pulse burst is defined by a
randomly varying parameter in which instantaneous minimum amplitude
thereof is not smaller than the maximum amplitude thereof by a
factor of one tenth-thousandth. The pulse burst repetition rate can
vary from about 0.01 to about 10,000 Hz. A mathematically definable
parameter can also be employed to define an amplitude envelope of
said pulse bursts.
[0017] By increasing a range of frequency components transmitted to
relevant cellular pathways, access to a large range of biophysical
phenomena applicable to known healing mechanisms, including
enhanced enzyme activity and growth factor and cytokine release, is
advantageously achieved.
[0018] According to an embodiment of the present invention, by
applying a random, or other high spectral density envelope, to a
pulse burst envelope of mono- or bi-polar rectangular or sinusoidal
pulses which induce peak electric fields between 10.sup.-6 and 10
volts per centimeter (V/cm), a more efficient and greater effect
can be achieved on biological healing processes applicable to both
soft and hard tissues in humans, animals and plants. A pulse burst
envelope of higher spectral density can advantageously and
efficiently couple to physiologically relevant dielectric pathways,
such as, cellular membrane receptors, ion binding to cellular
enzymes, and general transmembrane potential changes thereby
modulating angiogenesis and neovascularization.
[0019] By advantageously applying a high spectral density voltage
envelope as a modulating or pulse-burst defining parameter, power
requirements for such modulated pulse bursts can be significantly
lower than that of an unmodulated pulse. This is due to more
efficient matching of the frequency components to the relevant
cellular/molecular process. Accordingly, the dual advantages of
enhanced transmitting dosimetry to relevant dielectric pathways and
of decreasing power requirements are achieved.
[0020] A preferred embodiment according to the present invention
utilizes a Power Signal to Noise Ratio ("Power SNR") approach to
configure bioeffective waveforms and incorporates miniaturized
circuitry and lightweight flexible coils. This advantageously
allows a device that utilizes a Power SNR approach, miniaturized
circuitry, and lightweight flexible coils, to be completely
portable and if desired to be constructed as disposable and if
further desired to be constructed as implantable.
[0021] Specifically, broad spectral density bursts of
electromagnetic waveforms, configured to achieve maximum signal
power within a bandpass of a biological target, are selectively
applied to target pathway structures such as living organs,
tissues, cells and molecules. Waveforms are selected using a unique
amplitude/power comparison with that of thermal noise in a target
pathway structure. Signals comprise bursts of at least one of
sinusoidal, rectangular, chaotic and random wave shapes, have
frequency content in a range of about 0.01 Hz to about 100 MHz at
about 1 to about 100,000 bursts per second, and have a burst
repetition rate from about 0.01 to about 1000 bursts/second. Peak
signal amplitude at a target pathway structure such as tissue, lies
in a range of about 1 .mu.V/cm to about 100 mV/cm. Each signal
burst envelope may be a random function providing a means to
accommodate different electromagnetic characteristics of healing
tissue. A preferred embodiment according to the present invention
comprises about 0.1 to about 100 millisecond pulse burst comprising
about 1 to about 200 microsecond symmetrical or asymmetrical pulses
repeating at about 0.1 to about 100 kilohertz within the burst. The
burst envelope is a modified 1/f function and is applied at random
repetition rates between about 0.1 and about 1000 Hz. Fixed
repetition rates can also be used between about 0.1 Hz and about
1000 Hz. An induced electric field from about 0.001 mV/cm to about
100 mV/cm is generated. Another embodiment according to the present
invention comprises an about 0.01 millisecond to an about 10
millisecond burst of high frequency sinusoidal waves, such as 27.12
MHz, repeating at about 1 to about 100 bursts per second. An
induced electric field from about 0.001 mV/cm to about 100 mV/cm is
generated. Resulting waveforms can be delivered via inductive or
capacitive coupling.
[0022] It is an object of the present invention to provide
modulation of electromagnetically sensitive regulatory processes at
the cell membrane and at junctional interfaces between cells.
[0023] It is another object of the present invention to provide an
electromagnetic method of treatment of living cells and tissues
comprising a broad-band, high spectral density electromagnetic
field.
[0024] It is a further object of the present invention to provide
an electromagnetic method of treatment of living cells and tissues
comprising amplitude modulation of a pulse burst envelope of an
electromagnetic signal that will induce coupling with a maximum
number of relevant EMF-sensitive pathways in cells or tissues.
[0025] It is another object of the present invention to provide
increased blood flow to affected tissues by modulating angiogenesis
and neovascularization.
[0026] It is another object of the present invention to provide
increased blood flow to enhance viability, growth, and
differentiation of implanted cells, such as stem cells, tissues and
organs.
[0027] It is another object of the present invention to provide
increased blood flow in cardiovascular diseases by modulating
angiogenesis and neovascularization.
[0028] It is another object of the present invention to improve
micro-vascular blood perfusion and reduced transudation.
[0029] It is a another object of the present invention to provide a
treatment of maladies of the bone and other hard tissue by
modulating angiogenesis and neovascularization.
[0030] It is a still further object of the present invention to
provide a treatment of edema and swelling of soft tissue by
increased blood flow through modulation of angiogenesis and
neovascularization.
[0031] It is another object of the present invention to provide an
electromagnetic method of treatment of living cells and tissues
that can be used for repair of damaged soft tissue.
[0032] It is yet another object of the present invention to
increase blood flow to damaged tissue by modulation of vasodilation
and stimulating neovascularization.
[0033] It is a yet further object of the present invention to
provide an apparatus for modulation of angiogenesis and
neovascularization that can be operated at reduced power levels and
still possess benefits of safety, economics, portability, and
reduced electromagnetic interference.
[0034] It is an object of the present invention to configure a
power spectrum of a waveform by mathematical simulation by using
signal to noise ratio ("SNR") analysis to configure a waveform
optimized to modulate angiogenesis and neovascularization then
coupling the configured waveform using a generating device such as
ultra lightweight wire coils that are powered by a waveform
configuration device such as miniaturized electronic circuitry.
[0035] It is another object of the present invention to modulate
angiogenesis and neovascularization by evaluating Power SNR for any
target pathway structure such as molecules, cells, tissues and
organs of plants, animals and humans using any input waveform, even
if electrical equivalents are non-linear as in a Hodgkin-Huxley
membrane model.
[0036] It is another object of the present invention to provide a
method and apparatus for treating plants, animals and humans using
electromagnetic fields, selected by optimizing a power spectrum of
a waveform to be applied to a biochemical target pathway structure
to enable modulation of angiogenesis and neovascularization within
molecules, cells, tissues and organs of a plant, animal, and
human.
[0037] It is another object of the present invention to
significantly lower peak amplitudes and shorter pulse duration.
This can be accomplished by matching via Power SNR, a frequency
range in a signal to frequency response and sensitivity of a target
pathway structure such as a molecule, cell, tissue, and organ, of
plants, animals and humans to enable modulation of angiogenesis and
neovascularization.
[0038] The above and yet other objects and advantages of the
present invention will become apparent from the hereinafter set
forth Brief Description of the Drawings, Detailed Description of
the Invention, and Claims appended herewith.
BRIEF DESCRIPTION OF THE DRAWINGS
[0039] Preferred embodiments of the present invention will be
described below in more detail, with reference to the accompanying
drawings:
[0040] FIG. 1 is a flow diagram of a electromagnetic treatment
method for angiogenesis modulation of living tissues and cells
according to an embodiment of the present invention;
[0041] FIG. 2 is a view of control circuitry according to a
preferred embodiment of the present invention;
[0042] FIG. 3 is a block diagram of miniaturized circuitry
according to a preferred embodiment of the present invention;
[0043] FIG. 4 depicts a waveform delivered to a angiogenesis and
neovascularization target pathway structure according to a
preferred embodiment of the present invention.
DETAILED DESCRIPTION
[0044] Induced time-varying currents from PEMF or PRF devices flow
in a target pathway structure such as a molecule, cell, tissue, and
organ, and it is these currents that are a stimulus to which cells
and tissues can react in a physiologically meaningful manner. The
electrical properties of a target pathway structure affect levels
and distributions of induced current. Molecules, cells, tissue, and
organs are all in an induced current pathway such as cells in a gap
junction contact. Ion or ligand interactions at binding sites on
macromolecules that may reside on a membrane surface area voltage
dependent processes, that is electrochemical, that can respond to
an induced electromagnetic field ("E"). Induced current arrives at
these sites via a surrounding ionic medium. The presence of cells
in a current pathway causes an induced current ("J") to decay more
rapidly with time ("J(t)"). This is due to an added electrical
impedance of cells from membrane capacitance and time constants of
binding and other voltage sensitive membrane processes such as
membrane transport.
[0045] Equivalent electrical circuit models representing various
membrane and charged interface configurations, have been derived.
For example, in Calcium ("Ca.sup.2+") binding, the change in
concentration of bound Ca.sup.2+ at a binding site due to induced E
may be described in a frequency domain by an impedance expression
such as: 1 Z b ( ) = R ion + 1 i C ion
[0046] which has the form of a series resistance-capacitance
electrical equivalent circuit. Where .omega. is angular frequency
defined as 2.pi.f, where f is frequency, i=-1.sup.1/2,
Z.sub.b(.omega.) is the binding impedance, and R.sub.ion and
C.sub.ion are equivalent binding resistance and capacitance of an
ion binding pathway. The value of the equivalent binding time
constant, .tau..sub.ion=R.sub.ionC.sub.ion, is related to a ion
binding rate constant, k.sub.b, via
.tau..sub.ion=R.sub.ionC.sub.ion=- 1/k.sub.b. Thus, the
characteristic time constant of this pathway is determined by ion
binding kinetics.
[0047] Induced E from a PEMF or PRF signal can cause current to
flow into an ion binding pathway and affect the number of Ca.sup.2+
ions bound per unit time. An electrical equivalent of this is a
change in voltage across the equivalent binding capacitance
C.sub.ion, which is a direct measure of the change in electrical
charge stored by C.sub.ion. Electrical charge is directly
proportional to a surface concentration of Ca.sup.2+ ions in the
binding site, that is storage of charge is equivalent to storage of
ions or other charged species on cell surfaces and junctions.
Electrical impedance measurements, as well as direct kinetic
analyses of binding rate constants, provide values for time
constants necessary for configuration of a PMF waveform to match a
bandpass of target pathway structures. This allows for a required
range of frequencies for any given induced E waveform for optimal
coupling to target impedance, such as bandpass.
[0048] Ion binding to regulatory molecules is a frequent EMF
target, for example Ca.sup.2+ binding to calmodulin ("CaM"). Use of
this pathway is based upon acceleration of wound repair, for
example bone repair, that involves modulation of growth factors
released in various stages of repair. Growth factors such as
platelet derived growth factor ("PDGF"), fibroblast growth factor
("FGF"), and epidermal growth factor ("EGF") are all involved at an
appropriate stage of healing. Angiogenesis and neovascularization
are also integral to wound repair and can be modulated by PMF. All
of these factors are Ca/CaM-dependent.
[0049] Utilizing a Ca/CaM pathway a waveform can be configured for
which induced power is sufficiently above background thermal noise
power. Under correct physiological conditions, this waveform can
have a physiologically significant bioeffect.
[0050] Application of a Power SNR model to Ca/CaM requires
knowledge of electrical equivalents of Ca.sup.2+ binding kinetics
at CaM. Within first order binding kinetics, changes in
concentration of bound Ca.sup.2+ at CaM binding sites over time may
be characterized in a frequency domain by an equivalent binding
time constant, .tau..sub.ion=R.sub.ionC.sub.ion, where R.sub.ion
and C.sub.ion are equivalent binding resistance and capacitance of
the ion binding pathway. .tau..sub.ion is related to a ion binding
rate constant, k.sub.b, via .tau..sub.ion=R.sub.ionC.sub.ion=1/k.-
sub.b. Published values for k.sub.b can then be employed in a cell
array model to evaluate SNR by comparing voltage induced by a PRF
signal to thermal fluctuations in voltage at a CaM binding site.
Employing numerical values for PMF response, such as
V.sub.max=6.5.times.10.sup.-7 sec.sup.-1, [Ca.sup.2+]=2.5 .mu.M,
K.sub.D=30 .mu.M, [Ca.sup.2+CaM]=K.sub.D([Ca.sup.2+]+[CaM]), yields
k.sub.b=665 sec.sup.-1 (.tau..sub.ion=1.5 msec). Such a value for
.tau..sub.ion can be employed in an electrical equivalent circuit
for ion binding while power SNR analysis, can be performed for any
waveform structure.
[0051] According to an embodiment of the present invention a
mathematical model can be configured to assimilate that thermal
noise is present in all voltage dependent processes and represents
a minimum threshold requirement to establish adequate SNR. Power
spectral density, S.sub.n(.omega.), of thermal noise can be
expressed as:
S.sub.n(.omega.)=4kT Re[Z.sub.M(x,.omega.)]
[0052] where Z.sub.M(x,.omega.) is electrical impedance of a target
pathway structure, x is a dimension of a target pathway structure
and Re denotes a real part of impedance of a target pathway
structure. Z.sub.M(x,.omega.) can be expressed as: 2 Z M ( x , ) =
[ R e + R i + R g ] tanh ( x )
[0053] This equation clearly shows that electrical impedance of the
target pathway structure, and contributions from extracellular
fluid resistance ("R.sub.e"), intracellular fluid resistance
("R.sub.i") and intermembrane resistance ("R.sub.g") which are
electrically connected to a target pathway structure, all
contribute to noise filtering.
[0054] A typical approach to evaluation of SNR uses a single value
of a root mean square (RMS) noise voltage. This is calculated by
taking a square root of an integration of S.sub.n(.omega.)=4kT
Re[Z.sub.M(x,.omega.)] over all frequencies relevant to either
complete membrane response, or to bandwidth of a target pathway
structure. SNR can be expressed by a ratio: 3 SNR = V M ( ) RMS
[0055] where .vertline.V.sub.M(.omega.).vertline. is maximum
amplitude of voltage at each frequency as delivered by a chosen
waveform to the target pathway structure.
[0056] An embodiment according to the present invention comprises a
pulse burst envelope having a high spectral density, so that the
effect of therapy upon the relevant dielectric pathways, such as,
cellular membrane receptors, ion binding to cellular enzymes and
general transmembrane potential changes, is enhanced. Accordingly
by increasing a number of frequency components transmitted to
relevant cellular pathways, a large range of biophysical phenomena,
such as modulating growth factor and cytokine release and ion
binding at regulatory molecules, applicable to known healing
mechanisms is accessible. According to an embodiment of the present
invention applying a random, or other high spectral density
envelope, to a pulse burst envelope of mono- or bi-polar
rectangular or sinusoidal pulses inducing peak electric fields,
between about 10.sup.-6and about 100 V/cm, produces a greater
effect on biological healing processes applicable to both soft and
hard tissues.
[0057] According to yet another embodiment of the present invention
by applying a high spectral density voltage envelope as a
modulating or pulse-burst defining parameter, power requirements
for such amplitude modulated pulse bursts can be significantly
lower than that of an unmodulated pulse burst containing pulses
within a similar frequency range. This is due to a substantial
reduction in duty cycle within repetitive burst trains brought
about by imposition of an irregular, and preferably random,
amplitude onto what would otherwise be a substantially uniform
pulse burst envelope. Accordingly, the dual advantages, of enhanced
transmitted dosimetry to the relevant dielectric pathways and of
decreased power requirement are achieved.
[0058] Referring to FIG. 1, wherein FIG. 1 is a flow diagram of a
method for delivering electromagnetic signals to angiogenesis and
neovascularization target pathway structures such as ions and
ligands of plants, animals, and humans for therapeutic and
prophylactic purposes according to an embodiment of the present
invention. A mathematical model having at least one waveform
parameter is applied to configure at least one waveform to be
coupled to a angiogenesis and neovascularization target pathway
structure such as ions and ligands (Step 101). The configured
waveform satisfies a SNR or Power SNR model so that for a given and
known angiogenesis and neovascularization target pathway structure
it is possible to choose at least one waveform parameter so that a
waveform is detectable in the angiogenesis and neovascularization
target pathway structure above its background activity (Step 102)
such as baseline thermal fluctuations in voltage and electrical
impedance at a target pathway structure that depend upon a state of
a cell and tissue, that is whether the state is at least one of
resting, growing, replacing, and responding to injury. A preferred
embodiment of a generated electromagnetic signal is comprised of a
burst of arbitrary waveforms having at least one waveform parameter
that includes a plurality of frequency components ranging from
about 0.01 Hz to about 100 MHz wherein the plurality of frequency
components satisfies a Power SNR model (Step 102). A repetitive
electromagnetic signal can be generated for example inductively or
capacitively, from said configured at least one waveform (Step
103). The electromagnetic signal is coupled to a angiogenesis and
neovascularization target pathway structure such as ions and
ligands by output of a coupling device such as an electrode or an
inductor, placed in close proximity to the target pathway structure
(Step 104). The coupling enhances modulation of binding of ions and
ligands to regulatory molecule in living tissues and cells.
[0059] FIG. 2 illustrates a preferred embodiment of an apparatus
according to the present invention. A miniature control circuit 201
is coupled to an end of at least one connector 202 such as wire.
The opposite end of the at least one connector is coupled to a
generating device such as a pair of electrical coils 203. The
miniature control circuit 201 is constructed in a manner that
applies a mathematical model that is used to configure waveforms.
The configured waveforms have to satisfy a SNR or Power SNR model
so that for a given and known angiogenesis and neovascularization
target pathway structure, it is possible to choose waveform
parameters that satisfy SNR or Power SNR so that a waveform is
detectable in the angiogenesis and neovascularization target
pathway structure above its background activity. A preferred
embodiment according to the present invention applies a
mathematical model to induce a time-varying magnetic field and a
time-varying electric field in a angiogenesis and
neovascularization target pathway structure such as ions and
ligands comprising about 10 to about 100 msec bursts of about 1 to
about 100 microsecond rectangular pulses repeating at about 0.1 to
about 10 pulses per second. Peak amplitude of the induced electric
field is between about 1 uV/cm and about 100 mV/cm, varied
according to a modified 1/f function where f=frequency. A waveform
configured using a preferred embodiment according to the present
invention may be applied to a angiogenesis, and neovascularization
target pathway structure such as ions and ligands for a preferred
total exposure time of under 1 minute to 240 minutes daily. However
other exposure times can be used. Waveforms configured by the
miniature control circuit 201 are directed to a generating device
203 such as electrical coils via connector 202. The generating
device 203 delivers a pulsing magnetic field configured according
to a mathematical model, that can be used to provide treatment to a
angiogenesis and neovascularization target pathway structure such
as a heart in a chest 204. The miniature control circuit applies a
pulsing magnetic field for a prescribed title and can automatically
repeat applying the pulsing magnetic field for as many applications
as are needed in a given time period, for example 10 times a day. A
preferred embodiment according to the present invention can be
positioned to treat the heart in a chest 204 by a positioning
device. Coupling a pulsing magnetic field to a angiogenesis and
neovascularization target pathway structure such as ions and
ligands, therapeutically and prophylactically reduces inflammation
thereby reducing pain and promotes healing. When electrical coils
are used as the generating device 203, the electrical coils can be
powered with a time varying magnetic field that induces a time
varying electric field in a target pathway structure according to
Faraday's law. An electromagnetic signal generated by the
generating device 203 can also be applied using electrochemical
coupling, wherein electrodes are in direct contact with skin or
another outer electrically conductive boundary of a target pathway
structure. Yet in another embodiment according to the present
invention, the electromagnetic signal generated by the generating
device 203 can also be applied using electrostatic coupling wherein
an air gap exists between a generating device 203 such as an
electrode and a angiogenesis and neovascularization target pathway
structure such as ions and ligands. An advantage of the preferred
embodiment according to the present invention is that its ultra
lightweight coils and miniaturized circuitry allow for use with
common physical therapy treatment modalities and at any body
location for which pain relief and healing is desired. An
advantageous result of application of the preferred embodiment
according to the present invention is that a living organism's
angiogenesis and neovascularization can be maintained and
enhanced.
[0060] FIG. 3 depicts a block diagram of a preferred embodiment
according to the present invention of a miniature control circuit
300. The miniature control circuit 300 produces waveforms that
drive a generating device such as wire coils described above in
FIG. 2. The miniature control circuit can be activated by any
activation means such as an on/off switch. The miniature control
circuit 300 has a power source such as a lithium an output voltage
of 3.3 V but other voltages can be used. In another embodiment
according to the present invention the power source can be an
external power source such as an electric current outlet such as an
AC/DC outlet, coupled to the present invention for example by a
plug and wire. A switching power supply 302 controls voltage to a
micro-controller 303. A preferred embodiment of the
micro-controller 303 uses an 8 bit 4 MHz micro-controller 303 but
other bit MHz combination micro-controllers may be used. The
switching power supply 302 also delivers current to storage
capacitors 304. A preferred embodiment of the present invention
uses storage capacitors having a 220 uF output but other outputs
can be used. The storage capacitors 304 allow high frequency pulses
to be delivered to a coupling device such as inductors (Not Shown).
The micro-controller 303 also controls a pulse shaper 305 and a
pulse phase timing control 306. The pulse shaper 305 and pulse
phase timing control 306 determine pulse shape, burst width, burst
envelope shape, and burst repetition rate. An integral waveform
generator, such as a sine wave or arbitrary number generator can
also be incorporated to provide specific waveforms. A voltage level
conversion sub-circuit 308 controls an induced field delivered to a
target pathway structure. A switching Hexfet 308 allows pulses of
randomized amptitude to be delivered to output 309 that routes a
waveform to at least one coupling device such as an inductor. The
micro-controller 303 can also control pathway structure such as a
molecule, cell, tissue, and organ. The miniature control circuit
300 can be constructed to apply a pulsing magnetic field for a
prescribed time and to automatically repeat applying the pulsing
magnetic field for as many applications as are needed in a given
time period, for example 10 times a day. A preferred embodiment
according to the present invention uses treatments times of about
10 minutes to about 30 minutes.
[0061] Referring to FIG. 4 an embodiment according to the present
invention of a waveform 400 is illustrated. A pulse 401 is repeated
within a burst 402 that has a finite duration 403. The duration 403
is such that a duty cycle which can be defined, as a ratio of burst
duration to signal period is between about 1 to about 10.sup.-5. A
preferred embodiment according to the present invention utilizes
pseudo rectangular 10 microsecond pulses for pulse 401 applied in a
burst 402 for about 10 to about 50 msec having a modified 1/f
amplitude envelope 404 and with a finite duration 403 corresponding
to a burst period of between about 0.1 and about 10 seconds.
EXAMPLE 1
[0062] The Power SNR approach for PMF signal configuration has been
tested experimentally on calcium dependent myosin phosphorylation
in a standard enzyme assay. The cell-free reaction mixture was
chosen for phosphorylation rate to be linear in time for several
minutes, and for sub-saturation Ca.sup.2+ concentration. This opens
the biological window for Ca.sup.2+/CaM to be EMF-sensitive. This
system is not responsive to PMF at levels utilized in this study if
Ca.sup.2+ is at saturation levels with respect to CaM, and reaction
is not slowed to a minute time range. Experiments were performed
using myosin light chain ("MLC") and myosin light chain kinase,
("MLCK") isolated from turkey gizzard. A reaction mixture consisted
of a basic solution containing 40 mM Hepes buffer, pH 7.0; 0.5 mM
magnesium acetate; 1 mg/ml bovine serum albumin, 0.1% (w/v) Tween
80; and 1 mM EGTA12. Free Ca.sup.2+ was varied in the 1-7 .mu.M
range. Once Ca.sup.2+ buffering was established, freshly prepared
70 nM CaM, 160 nM MLC and 2 nM MLCK were added to the basic
solution to form a final reaction mixture. The low MLC/MLCK ratio
allowed linear time behavior in the minute time range. This
provided reproducible enzyme activities and minimized pipetting
time errors.
[0063] The reaction mixture, was freshly prepared daily for each
series of experiments and was aliquoted in 100 .mu.L portions, into
1.5 ml Eppendorf tubes. All Eppendorf tubes, containing reaction
mixture were kept at 0.degree. C. then transferred to a specially
designied water bath maintained at 37.+-.0.1.degree. C. by constant
perfusion of water prewarmed by passage through a Fisher Scientific
model 900 heat exchanger. Temperature was monitored with a
thermistor probe such as a Cole-Parmer model 8110-20, immersed in
one Eppendorf tube during all experiments. Reaction was initiated
with 2.5 .mu.M 3.2P ATP, and was stopped with Laemmli Sample Buffer
solution containing 30 .mu.M EDTA. A minimum of five blank samples
were counted in each experiment. Blanks comprised a total assay
mixture minus one of the active components Ca.sup.2+, CaM, MLC or
MLCK. Experiments for which blank counts were higher than 300 cpm
were rejected. Phosphorylation was allowed to proceed for 5 min and
was, evaluated by counting 32P incorporated in MLC using a TM
Analytic model 5303 Mark V liquid scintillation counter.
[0064] The signal comprised repetitive bursts of a high frequency
waveform. Amplitude was maintained constant at 0.2 G and repetition
rate was 1 burst/sec for all exposures. Burst duration varied from
65 .mu.sec to 1000 .mu.sec based upon projections of Power SNR
analysis which showed that optimal Power SNR would be achieved as
burst duration approached 500 .mu.sec. The results are shown in
FIG. 7 wherein burst width 701 in .mu.sec is plotted on the x-axis
and Myosin Phosphorylation 702 as treated/sham is plotted on the
y-axis. It can be seen that the PMF effect on Ca.sup.2+ binding to
CaM approaches its maximum at approximately 500 .mu.sec, just as
illustrated by the Power SNR model.
[0065] These results confirm that a PMF signal, configured
according to an embodiment of the present invention, would
maximally increase myosin phosphorylation for burst durations
sufficient to achieve optimal Power SNR for a given magnetic field
amplitude.
EXAMPLE 2
[0066] According to an embodiment of the present, invention use of
a Power SNR model was further verified in an in vivo wound repair
model. A rat wound model has been well characterized both
biomechanically and biochemically, and was used in this study.
Healthy, young adult male Sprague Dawley rats weighing more than
300 grams were utilized.
[0067] The animals were anesthetized with an intraperitoneal dose
of Ketamine 75 mg/kg and Medetomidine 0.5 mg/kg. After adequate
anesthesia had been achieved, the dorsum was shaved, prepped with a
dilute betadine/alcohol solution, and draped using sterile
technique. Using a #10 scalpel, an 8-cm linear incision was
performed through the skin down to the fascia on the dorsum of each
rat. The wound edges were bluntly dissected to break any remaining
dermal fibers, leaving an open wound approximately 4 cm in
diameter. Hemostasis was obtained with applied pressure to avoid
any damage to the skin edges. The skin edges were then closed with
a 4-0 Ethilon running suture. Post-operatively, the animals
received Buprenorphine 0.1-0.5 mg/kg, intraperitoneal. They were
placed in individual cages and received food and water ad
libitum.
[0068] PMF exposure comprised two pulsed radio frequency waveforms.
The first was a standard clinical PRF signal comprising a 65
.mu.sec burst of 27.12 MHz sinusoidal waves at 1 Gauss, amplitude
and repeating at 600 bursts/sec. The second was a PRF signal
reconfigured according to an embodiment of the present invention.
For this signal burst duration was increased to 2000 .mu.sec and
the amplitude and repetition rate were reduced to 0.2 G and 5
bursts/sec respectively. PRF was applied for 30 minutes twice
daily.
[0069] Tensile strength was performed immediately after wound
excision. Two 1 cm width strips of skin were transected
perpendicular to the scar from each sample and used to measure the
tensile strength in kg/mm.sup.2. The strips were excised from the
same area in each rat to assure consistency of measurement. The
strips were then mounted on a maximum force generated before the
wound pulled apart was recorded. The final tensile strength for
comparison was determined by taking the average of the maximum load
in kilograms per mm.sup.2 of the two strips from the same
wound.
[0070] The results showed average tensile strength for the 65
.mu.sec 1 Gauss PRF signal was 19.3.+-.4.3 kg/mm.sup.2 for the
exposed group versus 13.0.+-.3.5 kg/mm.sup.2 for the control group
(p<0.01), which is a 48% increase. In contrast, the average
tensile strength for the 2000 .mu.sec 0.2 Gauss PRF signal,
configured according to an embodiment of the present invention
using a Power SNR model was 21.2.+-.5.6 kg/mm.sup.2 for the treated
group versus 13.7.+-.4.1 kg/mm.sup.2 (p<0.01) for the control
group, which is a 54% increase. The results for the two signals
were not significantly different from each other.
[0071] These results demonstrate that an embodiment of the present
invention allowed a new PRF signal to be configured that could be
produced with significantly lower power. The PRF signal configured
according to an embodiment of the present invention, accelerated
would repair in the rat model in a low power manner versus that for
a clinical PRF signal which accelerated wound repair but required
more than two orders of magnitude more power to produce.
EXAMPLE 3
[0072] In this example Jurkat cells react to PMF stimulation of a
T-cell receptor with cell cycle arrest and thus behave like normal
T-lymphocytes stimulated by antigens at the T-cell receptor such as
anti-CD3. For example in bone healing, results have shown both 60
Hz and PEMF fields decrease DNA synthesis of Jurkat cells, as is
expected since PMF interacts with the T-cell receptor in the
absence of a costimulatory signal. This is consistent with an
anti-inflammatory response, as has been observed in clinical
applications of PMF stimuli. The PEMF signal is more effective. A
dosimetry analysis performed according to an embodiment of the
present invention demonstrates why both signals are effective and
why PEMF signals have a greater effect than 60 Hz signals on Jurkat
cells in the most EMF-sensitive growth stage.
[0073] Comparison of dosimetry from the two signals employed
involves evaluation of the ratio of the Power spectrum of the
thermal noise voltage that is Power SNR, to that of the induced
voltage at the EMF-sensitive target pathway structure. The target
pathway structure used is ion binding at receptor sites on Jurkat
cells suspended in 2 mm of culture medium. The average peak
electric field at the binding site from a PEMF signal comprising 5
msec burst of 200 .mu.sec pulses repeating at 15/sec, was 1 mV/cm,
while for a 60 Hz signal it was 50 .mu.V/cm.
EXAMPLE 4
[0074] In this example electromagnetic field energy was used to
stimulate neovascularization in an in vivo model. Two different
signal were employed, one configured according to prior art and a
second configured according to an embodiment of the present
invention.
[0075] One hundred and eight Sprague-Dawley male rats weighing
approximately 300 grams each, were equally divided into nine
groups. All animals were anesthetized with a mixture of
ketamine/acepromazine/Stadol at 0.1 cc/g. Using sterile surgical
techniques, each animal had a 12 cm to 14 cm segment of tail artery
harvested using microsurgical tehnique. The artery was flushed with
60 U/ml of heparinized saline to remove any blood or emboli. These
tail vessels, with an average diameter of 0.4 mm to 0.5 mm, were
then sutured to the transected proximal and distal segments of the
right femoral artery using two end-to-end anastomoses, creating a
femoral arterial loop. The resulting loop was then placed in a
subcutaneous pocket created over the animal's abdominal wall/groin
musculature, and the groin incision was closed with 4-0 Ethilon.
Each animal was then randomly placed into one of nine groups:
groups 1 to 3 (controls), these rats received no electromagnetic
field treatments and were killed at 4, 8, and 12 weeks; groups 4 to
6, 30 min. treatments twice a day to using 0.1 gauss
electromagnetic fields for 4, 8, and 12 weeks (animals were killed
at 4, 8, and 12 weeks, respectively); and groups 7 to 9, 30 min.
treatments twice a day using 2.0 gauss electromagnetic fields for
4, 8, and 12 weeks (animals were killed at 4, 8, and 12 weeks,
respectively).
[0076] Pulsed electromagnetic energy was as applied to the treated
groups using a device constructed according to an embodiment of the
present invention. Animals in the experimental groups were treated
for 30 minutes twice a day at either 0.1 gauss or 2.0 gauss, using
short pulses (2 msec to 20 msec) 27.12 MHz. Animals were positioned
on top of the applicator head and confined to ensure that treatment
was properly applied. The rats were reanesthetized with
ketamine/acepromazine/Stadol intraperitoneally and 100 U/kg of
heparin intravenously. Using the previous groin incision the
femoral artery was identified and checked for patency. The
femoral/tail artery loop was then isolated proximally and distally
from the anastomoses sites, and the vessel was clamped off. Animals
were then killed. The loop was injected with saline followed by 0.5
cc to 1.0 cc of colored latex through a 25-gauge, cannula and
clamped. The overlying abdominal skin was carefully resected, and
the arterial loop was exposed. Neovascularization was quantified by
measuring the surface area covered by new blood-vessel formation
delineated by the intraluminal latex. All results were analyzed
using the SPSS statistical analysis package.
[0077] The most noticeable difference in neovascularization between
treated versus untreated rats occurred at week 4. At that time, no
new vessel formation was found among controls, however, each of the
treated groups had similar statistically significant evidence of
neovascularization at 0 cm2 versus 1.42.+-.0.80 cm2 (p<0.001).
These areas appeared as a latex blush segmentally distributed along
the sides of the arterial loop. At 8 weeks, controls began to
demonstrate neovascularization measured at 0.7.+-.0.82 cm2. Both
treated groups at 8 weeks again had approximately equal
statistically significant (p<0.001) outcroppings of blood
vessels of 3.57.+-.1.82 cm2 for the 0.1 gauss group and of
3.77.+-.1.82 cm2 for the 2.0 gauss group. At 12 weeks, animals in
the control group displayed 1.75.+-.0.95 cm2 of neovascularization,
whereas the 0.1 gauss group demonstrated 5.95.+-.3.25 cm2, and the
2.0 gauss group showed 6.20.+-.3.95 cm2 of arborizing vessels.
Again, both treated groups displayed comparable statistically
significant findings (p<0.001) over controls.
[0078] These experimental findings demonstrate that electromagnetic
field stimulation of an isolated arterial loop according to an
embodiment of the present invention increases the amount of
quantifiable neovascularization in an in vivo rat model. Increased
angiogenesis was demonstrated in each of the treated groups at each
of the sacrifice dates. No differences were found between the
results of the two gauss levels tested as predicted by the
teachings of the present invention.
[0079] Having described embodiments, for an apparatus and a method
for delivering electromagnetic treatment to human, animal and plant
molecules, cells, tissue and organs, it is noted that modifications
and variations can be made by person skilled in the art in light of
the above teachings. It is therefore to be understood that changes
may be made in the particular embodiments of the invention
disclosed which are within the scope and spirit of the invention as
defined by the appended claims.
* * * * *