U.S. patent application number 14/085235 was filed with the patent office on 2014-09-04 for drug delivery by carbon nanotube arrays.
This patent application is currently assigned to CALIFORNIA INSTITUTE OF TECHNOLOGY. The applicant listed for this patent is CALIFORNIA INSTITUTE OF TECHNOLOGY. Invention is credited to Adrianus Indrat Aria, Masoud Beizai, Morteza Gharib.
Application Number | 20140248216 14/085235 |
Document ID | / |
Family ID | 45770898 |
Filed Date | 2014-09-04 |
United States Patent
Application |
20140248216 |
Kind Code |
A1 |
Gharib; Morteza ; et
al. |
September 4, 2014 |
DRUG DELIVERY BY CARBON NANOTUBE ARRAYS
Abstract
The invention generally relates to carbon nanotube based drug
delivery methods, devices, and compositions. More particularly, the
invention relates to controlled drug delivery using anchored carbon
nanotube arrays.
Inventors: |
Gharib; Morteza; (Altadena,
CA) ; Aria; Adrianus Indrat; (Los Angeles, CA)
; Beizai; Masoud; (Altadena, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
CALIFORNIA INSTITUTE OF TECHNOLOGY |
Pasadena |
CA |
US |
|
|
Assignee: |
CALIFORNIA INSTITUTE OF
TECHNOLOGY
Pasadena
CA
|
Family ID: |
45770898 |
Appl. No.: |
14/085235 |
Filed: |
November 20, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13224287 |
Sep 1, 2011 |
8784373 |
|
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14085235 |
|
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61379701 |
Sep 2, 2010 |
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Current U.S.
Class: |
424/9.2 ;
424/423; 427/2.14 |
Current CPC
Class: |
A61L 29/16 20130101;
A61L 2300/422 20130101; A61K 31/352 20130101; A61K 31/337 20130101;
A61K 49/0004 20130101; A61K 49/0095 20130101; A61K 47/6929
20170801; A61K 9/5192 20130101; B82Y 40/00 20130101; A61K 47/02
20130101; A61L 29/103 20130101; B82Y 5/00 20130101 |
Class at
Publication: |
424/9.2 ;
424/423; 427/2.14 |
International
Class: |
A61K 47/02 20060101
A61K047/02; A61K 49/00 20060101 A61K049/00; A61K 9/51 20060101
A61K009/51; A61K 31/337 20060101 A61K031/337 |
Claims
1. The method of claim 1, wherein depositing the agent to the
plurality of carbon nanotubes comprises: (i) mixing the agent in a
low surface tension solvent forming a solution of the agent; (ii)
coating the plurality of carbon nanotubes with the solution of the
agent; and (iii) drying the coated plurality of carbon nanotubes,
thereby removing the solvent while leaving the agent associated
with the plurality of carbon nano tubes.
2. The method of claim 1, wherein the low surface tension solvent
is an alcohol.
3. The method of claim 2, wherein the alcohol is ethanol.
4. An implantable drug delivery device, comprising: (a) an
implantable device; (b) an array of carbon nanotubes anchored on
the implantable device; and (c) an agent deposited on the array of
carbon nanotubes, wherein the agent is not covalently bound to the
carbon nanotubes.
5. The implantable drug delivery device of claim 4, wherein the
agent is a pharmaceutical agent capable of providing a therapeutic
effect.
6. The implantable drug delivery device of claim 4, wherein the
agent is a diagnostic agent capable of providing a detectable
signal or image indicating a biologically relevant state of the
subject.
7. The implantable drug delivery device of claim 4, wherein the
agent comprises an aromatic moiety.
8. A method for monitoring in situ delivery of an agent,
comprising: (a) providing a plurality of carbon nanotubes
non-covalently associated thereon a pharmaceutical agent and a
second agent capable of exhibiting a spatially detectable signal;
(b) placing the plurality of carbon nanotubes at a target location
in the patient's body; and (c) measuring the detectable signal
exhibited from the second agent to monitor the deliver of the
pharmaceutical agent in situ.
9. The method of claim 8, wherein the agent is a pharmaceutical
agent comprises an aromatic moiety.
Description
PRIORITY CLAIMS AND CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional application of U.S.
application Ser. No. 13/224,287, filed Sep. 1, 2011, which claims
the benefit of priority from U.S. Provisional Application Ser. No.
61/379,701, filed Sep. 2, 2010, the entire content of which is
incorporated herein by reference in its entirety.
TECHNICAL FIELD OF THE INVENTION
[0002] The invention generally relates to carbon nanotube based
drug delivery methods, devices, and compositions. More
particularly, the invention relates to controlled drug delivery
using anchored carbon nanotube arrays.
BACKGROUND OF THE INVENTION
[0003] Targeted, localized and controlled drug delivery remains a
major challenge. In many cases, efficacy of a drug can be improved
and the risks of side effects reduced if the therapy is
administered locally and/or continuously, rather than through
conventional oral ingestion or injection, which produce burst
releases. In some cases, dose-limiting toxicity levels are caused
by agent losses in vascular travel during transplant procedures.
Continuous and accurate local dosing is highly desirable, but
remains a major challenge, particularly in the cardiovascular field
where requirements for a material's biocompatibility and dosing
control are stringent.
[0004] Current diffusion-based drug-delivery platforms suffer from
very slow mass-transfer process. The published reports indicate
involvement of solid/solid diffusion as well as channel (e.g.,
tubule) and solvent-help (e.g., capillary, osmotic) mechanisms but
not convection. (Tepe, et al. 2007 Touch Briefings
2007--Interventional Cardiology, pp. 61-63; Scheller, et al. 2004
Circulation 110:810-814; Diaz, et al. 2005 J Biol Chem
280:3928-3937; Creel, et al. 2000 Circ Res 86:879-884; Lovich, et
al. 2001 J Pharm Sci 90:1324-1335; Zilberman, et al. 2008 J Biomed
Mater Res 84A:313-323; Davies 1997 N Engl J Med 336:1312-1314;
Arakawa, et al. 2002 Arterioscler Thromb Vase Biol 22:1002-1007;
Parekh, et al. 1997 Gen Pharmac 29:167-172; Heam, et al. 2009
Nature 458:367-371; Celermajer 2002 European Heart Journal
Supplement F:F24-F28; Andersen, et al. 2006 BMC Clinical
Pharmacology, published online 13 Jan. 2006; Oreopoulos, et al.
2009 J Structural Biology 168:21-36; Panchagnula, et al. 2004 J
Pharm Sci 93:2-177-2183; Migliavacca, et al. 2007 Comput Methods
Biomech Biomed Engin 10:63-73; Arifin, et al. 2009 Pharmaceutical
Research, published online 29 Jul. 2009.) In percutaneous
transluminal angioplasty (PCTA) devices, for example, drug washout
and overdose remain serious challenges. For oncology applications,
for example, localized delivery of sufficient dose of anti-cancer
drug via targeted delivery is highly desirable.
[0005] In recent years, carbon nanotubes have attracted attention
due to their chemical, mechanical and geometric properties. Carbon
nanotubes (CNTs) are allotropes of carbon with a cylindrical
nanostructure and are members of the fullerene structural family.
Nanotubes are categorized as single-walled nanotubes and
multi-walled nanotubes. Carbon nanotubes are strong and stiff
materials in terms of tensile strength and elastic modulus
respectively. Various techniques have been developed to make
nanotubes, such as arc discharge, laser ablation, high-pressure
carbon monoxide, and chemical vapor deposition.
[0006] Researches have been reported on CNT-based drug delivery.
For example, a recent study was reported on drug delivery using
PEGylated-CNTs. (Liu, et al. 2008 Cancer Res. 68: (16), 6652). The
reported system is based on covalently attaching drug molecules to
PEGylated CNTs. Another research group used carbon nanotube-based
tumor-targeted drug delivery system, which consisted of a
functionalized CNTs linked to tumor-targeting modules as well as
prodrug modules. (Chen, et al. 2008 J Am Chem Soc 130:16778-16785.)
In both of the afore-mentioned approaches, functionalization of the
CNTs is required, which presents a number of complications and
procedural drawbacks.
[0007] One reported example of angioplasty drug delivery is a PTCA
balloon coated with paclitaxel in an iopromide matrix. The balloon
is inflated for 30-second contact with vascular wall to allow the
matrix to dissolve and paclitaxel to migrate into the smooth muscle
cell. (Scheller, et al. 2004 Circulation 110:810-814.) Major
problems with this device include iopromide being hydrophilic and
an X-ray contrast agent. The first causes some drug loss to blood
stream (although claimed to be about 6%) and the second leads to
adverse reactions for some patients. Furthermore, the balloon still
contains about 10% paclitaxel after detachment and only about 15%
remains in the plaque.
[0008] Another reported example of angioplasty drug delivery is a
system using vascular stents made of paclitaxel-eluting composite
fibers to deliver about 40% of drug, most of it over 30 days.
(Zilberman, et al. 2008 J Biomed Mater Res 84A:313-323.) Since the
main mass-transfer mechanism of this device is diffusion, the rate
is inherently slow. These drawbacks are in addition to the
well-documented risks and side effects associated with stents.
[0009] For angioplasty drug delivery monitoring, existing
technologies typically use a fluorescent dye administered
intravenously through a central venous line with a dose adapted to
body weight. (Detter, et al. 2007 Circulation 116:1007-1014;
Hattori, et al. 2009 Circ Cardiovasc Imaging 2:277-278; Hosono, et
al. 2010 Interact CardioVasc Thorac Surg 10:476-477; Tanaka, et al.
2009 J Thorac Cardiovasc Surg 138:133-140; Waseda, et al. 2009 JACC
Cardiovascular Imaging 2:604-612.). The illumination is provided by
near-infrared laser diodes with a typical output of 80 mW in a
field of view of 10 cm in diameter, eliminating tissue warming and
eye protection concerns. The fluorescence emission of the excited
dye is typically detected by an IR-CCD camera and digitized with a
frame grabber that provides real-time recording.
[0010] Therefore, there remains an urgent and unmet need for
improved drug delivery systems addressing the above-mentioned
shortcomings, particularly in the field of angioplasty drug
delivery.
SUMMARY OF THE INVENTION
[0011] The invention is based, in part, on the unique approach to
drug delivery using anchored carbon nanotube arrays. In particular,
the invention provides targeted, localized, and controlled drug
delivery using novel acnhored carbon nanotube arrays that carry
(e.g., non-covalently) the agent to be delivered, including
therapeutic and diagnostic agents.
[0012] In one aspect, the invention generally relates a method for
delivering an agent to a patient in situ. The method includes: (a)
providing a plurality of carbon nanotubes; (b) depositing the agent
to the plurality of carbon nanotubes such that the agent is
non-covalenty associated with the plurality of carbon nanotubes;
(c) placing the plurality of carbon nanotubes deposited with the
agent at a target location in the patient's body; and (d) allowing
the agent to diffuse from the plurality of carbon nanotubes,
thereby delivering the agent in situ.
[0013] In another aspect, the invention generally relates to an
implantable drug delivery device. The implantable drug delivery
device includes: (a) an implantable device; (b) an array of carbon
nanotubes anchored on the implantable drug delivery device; and (c)
an agent deposited on the array of carbon nanotubes, wherein the
agent is not covalently bound to the carbon nanotubes.
[0014] In yet another aspect, the invention generally relates to a
method for monitoring in situ delivery of an agent. The method
includes: (a) providing a plurality of carbon nanotubes
non-covalently associated thereon a pharmaceutical agent and a
second agent capable of exhibiting a spatially detectable signal;
(b) placing the plurality of carbon nanotubes at a target location
in the patient's body; and (c) measuring the detectable signal
exhibited from the second agent to monitor the delivery of the
pharmaceutical agent in situ.
[0015] The invention disclosed herein enables improved devices,
methods and compositions for treating a number of conditions where
targeted, localized, and controlled drug delivery are required.
BRIEF DESCRIPTION OF THE DRAWINGS
[0016] FIGS. 1(a)-(b) show an exemplary angioplasty-balloon fitted
with carbon nanotubes. FIG. 1(c) and FIG. 1(d) are enlarged views
of the carbon nanotubes.
[0017] FIG. 2 shows an exemplary carbon nanotube array prepared via
capillography methods.
[0018] FIG. 3 shows schematics of an array of anchored carbon
nanotubes (as tall as 1 mm) on a flexible substrate. (Sansom, et
al. 2008 Nanotechnology 19:035302; Sansom 2007 Ph.D. Thesis,
California Institute of Technology; Noca, et al. U.S. Pat. No.
7,491,628; Huang, et al. 2007 Nanotechnology 18:305302; Gharib, et
al. U.S. Pat. Appl. 20080145616A1.)
[0019] FIG. 4 shows exemplary surface-to-volume ratio comparisons
of bundled anchored NTs and conventional microneedles. For the same
total needle diameter of 50 .mu.m, a bundle of ACNT has almost 100
times larger surface area than a conventional microneedle, assuming
a modest CNT spacing of 100 nm center-to-center. The uppermost line
represents the surface-to-volume gain by decreasing the CNT spacing
by a factor of 2.
[0020] FIG. 5 shows a schematic drawing of anchored CNTs (as tall
as 1 mm) with drugs deposited in its interstices. (Zhou, et al.
2006 Nanotechnology 17:4845-4853.)
[0021] FIG. 6(a) shows a flutax-coated anchored CNT array. FIG.
6(b) shows an uncoated CNT array.
[0022] FIG. 7(a) shows exemplary results of recovered fluorescein
sodium and flutax-1 with the same initial concentration in DI
water. FIG. 7(b) shows exemplary results of recovered fluorescein
sodium with various initial concentrations in DI water.
[0023] FIGS. 8(a)-8(b) show a rubbing experiment using
3.times.1.times.5/128-inch glass microscopy slide rubbed on
substrates made of CNT-fitted latex sheet (FIG. 8(a)) and bare
latex sheet (FIG. 8(b)).
[0024] FIGS. 9(a)-(b) are exemplary images of
3.times.1.times.5/128-inch glass microscopy slides used in rubbing
experiment. A significant amount of flutax-1 has been washed-out
from the substrate made of bare latex sheet in FIG. 9(a) and almost
no flutax-1 washed-out from a substrate made of CNT-fitted latex
sheet in FIG. 9(b).
[0025] FIG. 10 shows exemplary recovered flutax-1 in pure ethanol
after the rubbing experiment, where the bare latex sheet shows that
no more flutax-1 remains (left) while much flutax-1 still remains
on the CNT-fitted latex sheet (right); tubes OD=16 mm.
[0026] FIG. 11(a) shows an exemplary PET angioplasty balloon
without CNTs. FIG. 11(b) shows a CNT-partially-fitted PET
angioplasty balloon. The black region on the CNT-partially-fitted
PET angioplasty balloon is an array of vertically aligned CNTs
anchored on latex, which cover the outer surface of the PET
balloon.
[0027] FIGS. 12(a)-(b) show an exemplary experiment setup using
agar gel inside a 860-.mu.m-ID, 1,450-.mu.m-OD glass capillary tube
with a substrate made of CNT-fitted latex sheet (FIG. 12(a)) and
bare latex sheet (FIG. 12(b)).
[0028] FIGS. 13(a)-(f) are an exemplary series of time lapse images
of mass transfer in 5 weight % agar gel, where the flutax-1 were
transferred via .about.300 .mu.m CNT-fitted latex sheet (left) and
bare latex sheet (right).
[0029] FIGS. 14(a)-(b) show an exemplary mass transfer profile of
flutax-1 in 5 weight % agar gel, where the flutax-1 were
transferred via .about.300 .mu.m CNT-fitted latex sheet (FIG.
14(a)) and bare latex sheet (FIG. 14(b)).
[0030] FIG. 15(a) shows a graph of location of 1% concentration vs.
time for a dried flutax-1 positioned as a shallow flat layer
(squares, smooth line) and within and anchored CNT array
(triangles, dashed line). The squares and triangles mark
experimental data and the dashed and smooth lines are numerical
solutions of the appropriate model. Schematic geometric
descriptions of the problem for a thin flutax layer and CNT array
are presented in FIG. 15(b) and FIG. 15(c), respectively.
DETAILED DESCRIPTION OF THE INVENTION
[0031] The invention is based in part on the discovery of a
universal drug-carrying and delivery platform capable of delivering
targeted, localized, and controlled amounts of drugs to a target
location within a patient's body. The devices, systems, methods and
compositions of the invention significantly remedy the difficult
dose control problems in current PCTA devices as well as the
complexity and costs associated with CNT functionalizations.
Different from the common methods that have been published earlier
(e.g., Tepe, et al. 2007 Touch Briefings 2007--Interventional
Cardiology, pp. 61-63; Scheller, et al. 2004 Circulation
110;810-814; Diaz, et al. 2005 J Biol Chem 280:3928-3937; Creel, et
al. 2008 Circ Res 86:879-884), the device of the invention
incorporates a drug on CNTs without having to functionalize the
CNTs. Conjugating drugs with functionalized CNTs is inherently
complicated and expensive. Additionally, the functionalizing
process renders the CNTs hydrophilic, such that the drug attached
to the CNTs is vulnerable to washout by the blood stream.
Additionally, as disclosed herein, the ability to penetrate the
arterial plaque cap and to deliver the drugs directly to the inner
part of the plaque is a major advantage of a CNT-fitted angioplasty
balloon over the conventional angioplasty-balloon or stent-fitted
angioplasty balloon.
[0032] CNTs, since their discovery, have been studied for various
applications ranging from highly adhesive layers, heat sinks,
structural composites, vibration damping layers, lithium storages,
to field emitters, due to their exceptional properties. (Iijima
1991 Nature 354; 56-58; Yurdumakan, et al. 2005 Chem Commun
30:3799-3801; Xu, et al. 2004 IEEE Inter Society Conference on
Thermal Phenomena 29:549-555; Veedu, et al. 2006 Nature Materials
5:457-462; Daraio, et al. 2004 Appl Phys Lett 85:5724-5726; Wang,
et al. 2006 Metals and Materials Int'l 12:413-416; Manohara, et al.
2005 J Vac Sci Tech B23:157-161). The strength and flexibility of
CNTs are just two of many exceptional physical properties of CNTs.
The accepted Young's elastic modulus for individual CNTs is
extraordinarily high, approaching 1 TPa (somewhat less for
multi-walled CNTs than single-walled CNTs), which is almost 5 times
higher than stainless steel. (Wong, et al. 1997 Science
277:1971-1975, Krishnan, et al. 1998 Phys Rev B58:14013-14019.)
[0033] This high Young's modulus of elasticity allows the anchored
CNTs to penetrate soft surfaces such as biological tissues upon
touching and consequently have the potential of delivering drugs
directly to the inner part of the tissue. According to the linear
elastic beam theory, the critical force for bucking of an axially
loaded clamped cylindrical beam in compression can be calculated
by:
F crit = .pi. 3 Ed 4 2 S 6 L 2 ##EQU00001##
where d is the diameter of the beam, L is the length, and E is the
Young's modulus. Assuming a bundle of several CNTs is clamped
together on the substrate, and typical diameter and length of a
bundle of CNTs are 1 .mu.m and 500 .mu.m, respectively, the
critical buckling force for each bundle of CNTs is around 0.484
.mu.N, or equivalent to 617 kPa. This critical buckling force for
each bundle of CNTs is great enough to overcome the required
fracture stress of the arterial plaque cap, for example, about
254.8 kPa. (Holzapfel, et al. 2004 J Bio Eng 126:657-665.) As it is
demonstrated herein, CNT bundles anchored on a flexible substrate
are sufficiently strong to penetrate soft tissues, e.g. arterial
wall or arterial plaque cap, when pressed upon and can deliver a
drug directly to the inner part of these targets.
[0034] Another extraordinary property of CNTs that makes them a
great fit for drug delivery is the hydrophobicity of CNTs. By
placing the drugs in the interstices of an array of CNTs, the
highly hydrophobic CNTs protect the drugs from being washed out by
any aqueous solution. Therefore, the CNT-fitted angioplasty
balloon, for example, would protect the drugs from being washed-out
during vascular travel to the target location.
[0035] Thus, in one aspect, the invention generally relates a
method for delivering an agent to a patient in situ. The method
includes: (a) providing a plurality of carbon nanotubes; (b)
depositing the agent to the plurality of carbon nanotubes such that
the agent is non-covalently associated with the plurality of carbon
nanotubes; (c) placing the plurality of carbon nanotubes deposited
with the agent at a target location in the patient's body; and (d)
allowing the agent to diffuse from the plurality of carbon
nanotubes, thereby delivering the agent in situ.
[0036] In another aspect, the invention generally related to an
implantable drug delivery device. The implantable drug delivery
device includes: (a) an implantable device; (b) an array of carbon
nanotubes anchored on the implantable device; and (c) an agent
deposited on the array of carbon nanotubes, wherein the agent is
not covalently bound to the carbon nanotubes.
[0037] In yet another aspect, the invention generally relates to a
method for monitoring in situ delivery of an agent. The method
includes: (a) providing a plurality of carbon nanotubes
non-covalently associated thereon a pharmaceutical agent and a
second agent capable of exhibiting a spatially detectable signal;
(b) placing the plurality of carbon nanotubes at a target location
in the patient's body; and (c) measuring the detectable signal
exhibited from the second agent to monitor the deliver of the
pharmaceutical agent in situ.
[0038] In certain embodiments, the agent is a pharmaceutical agent
capable of providing a therapeutic effect on the patient. Exemplary
pharmaceutical agents include: adrenaline (epinephrine),
amphetamine, atropine taxol (or its fluorescent derivative
flutax-1), and statins.
[0039] In certain other embodiments, the agent is a diagnostic
agent capable of providing a detectable signal or image indicating
a biologically relevant state of the subject. Exemplary diagnostic
agents include: electrochemical detectors of chemical warfare
agents, utilizing superior electrical properties of carbon
nanotubes.
[0040] In certain preferred embodiments, the agent comprises an
aromatic moiety (e.g., arenes and heteroarenes). The aromatic
moiety may include one or more heteroatoms selected from N, O and
S. The aromatic moiety may include a single aromatic ring or two or
more independent or fused rings. The agent may be mono-aromatic or
multi-aromatic.
[0041] In certain embodiments, the agent comprises a non-aromatic
extended .pi.-bond system.
[0042] Preferably, the agent may have a molecular weight from about
130 to about 1500 (e.g., from about 130 to about 1200, from about
130 to about 1000, from about 200 to about 1500, from about 200 to
about 1200, from about 200 to about 1000, from about 500 to about
1000).
[0043] In certain preferred embodiments, the plurality of carbon
nanotubes are in an array format and anchored on an implantable
device. In certain preferred embodiments, the plurality of carbon
nanotubes are not surface functionalized, e.g., for covalent
attachment.
[0044] In certain preferred embodiments, the implantable device is
an angioplasty balloon.
[0045] The angioplasty balloon is preferably anchored uniformly
with the plurality of carbon nanotubes, e.g., without surface
functionalization. The density of the plurality of carbon nanotubes
is typically from about 10.sup.10 nanotubes/cm.sup.2 to about
10.sup.11 nanotubes/cm.sup.2 (e.g., about 2.times.10.sup.10
nanotubes/cm.sup.2, 4.times.10.sup.10 nanotubes/cm.sup.2,
6.times.10.sup.10 nanotubes/cm.sup.2, 8.times.10.sup.10
nanotubes/cm.sup.2.) (FIGS. 1(a)-(d)).
[0046] In a preferred embodiment, the agent is an agent for
treating arterial plaque and the target location is interior wall
of a vascular lumen of the patient. In some embodiments, the target
location is inside an arterial plaque whereby at least some of the
carbon nanotubes penetrate the outer lining and enter the interior
of an arterial plaque.
[0047] The carbon nanotubes may be prepared by my suitable means,
e.g., by thermal chemical vapor deposition. In some embodiments,
the carbon nanotubes are multi-walled. In some embodiments, the
carbon nanotubes are single-walled. The desired specifications of
carbon nanotubes are dependent on the applications including the
requisite strength of the nanotubes.
[0048] In certain embodiments, the agent is deposited to the carbon
nanotubes by a method that includes: mixing the agent in a low
surface tension solvent forming a solution of the agent; coating
the plurality of carbon nanotubes with the solution of the agent;
and drying the coated plurality of carbon nanotubes, thereby
removing the solvent while leaving the agent associated with the
plurality of carbon nanotubes.
[0049] The low surface tension solvent may be any suitable solvent,
for example, an alcohol. In certain preferred embodiments, the low
surface tension solvent is pure (200 proof) ethanol or another
water-free alcohol.
[0050] The drug load is determined according to the requirements of
the application. Drug load may range from about 0.1 mg to about 20
mg (e.g., from about 0.1 mg to about 10 mg, from about 0.1 mg to
about 5 mg, from about 0.1 mg to about 2.5 mg, from about 1.0 mg to
about 20 mg, from about 1.0 mg to about 10 mg.)
[0051] CNTs can be formed in several ways, a simple way being a CVD
method with a catalyst-coated substrate (typically a few nanometers
of iron coated onto silicon wafer) prepared in advance and placed
in a tube furnace under saturated flow of carbon containing
feed-gas (e.g., ethylene) small portion of reducer feed-gas, e.g.
hydrogen, and elevated to proper temperature (e.g., 725.degree.
C.). Thermal CVD growth of CNTs in this way generates vertically
aligned CNTs on the growth substrate. (Sansom, et al. 2008
Nanotechnology 19:035302; Sansom 2007 Experimental Investigation on
Patterning of Anchored and Unanchored Aligned Carbon Nanotube Mats
by Fluid Immersion and Evaporation, Ph.D. Thesis, California
Institute of Technology.)
[0052] Various methods and techniques have been developed that
allow the preparation of anchored CNTs, including microscale fluid
transport and control techniques by "nanowicking" and by
self-assembly pattern formation and a method for controllable
anchoring of CNTs within polymer layers (Sansom, et al. 2008
Nanotechnology 19:035302; Sansom 2007 Experimental Investigation on
Patterning of Anchored and Unanchored Aligned Carbon Nanotube Mats
by Fluid Immersion and Evaporation, Ph.D. Thesis, California
Institute of Technology; Zhou, et al. 2006 Nanotechnology
17:4845-4853; U.S. Pat. No. 7,491,628 Noca, et al.; Huang, et al.
2007 Nanotechnology 18:305301; U.S. Pat. App. 20080145616A1 by
Gharib, et al.). Tall CNT arrays have been grown in arbitrary
patterns (e.g., bundles, rows, geometric shapes) on a substrate
with heights of over 1 mm (Bronikowski 2007 J Phys Chem C
111:17705-17712.)
[0053] In contrast to the length of CNTs that can be varied
relatively easily by varying either the thickness of catalyst layer
or the growth time, the packing density of an array of CNTs is
relatively harder to vary. One way to adjust the packing density of
the CNTs is by changing the timing and duration of the hydrogen
exposure during the CNT growth. (Nessim, et al. 2008 Nano Lett
8:3587-3593.) By this method, the packing density of as-grown CNTs
can be varied from 3.9.times.10.sup.9 to 4.9.times.10.sup.10
CNTs/cm.sup.2. Another approach is by compressing the as-grown CNTs
by external mechanical force or capillary force. (Wardle, et al.
2008 Adv Mater 20:2707-2714; Futaba, et al. 2006 Nature Material
5:987-994). By using an external mechanical force to compact the
as-grown CNTs, the packing density of CNTs can be increased up to
about 20%. By using a capillary force to collapse a pack of CNTs,
the packing density of CNTs can be increased up to about 50%, or
equivalent to the increase of packing density from
4.3.times.10.sup.11 to 8.3.times.10.sup.12 CNTs/cm.sup.2.
[0054] A major challenge for the vertically aligned CNTs is the
poor adhesion of the CNTs to their growth substrates, e.g., silicon
wafer. Although the mechanical properties of individual CNTs are
excellent, the collective properties of bulk vertically-aligned
CNTs have not been as good as expected, because of the weak bond
between the base of the CNTs and their growth substrates. A method
has been developed to overcome this problem by anchoring the CNTs
in a layer of flexible polymeric materials, e.g., PDMS
(polydimethylsiloxane), PMMA (poly(methyl methacrylate)), or latex,
(Sansom, et al. 2008 Nanotechnology 19:035302.) As-grown CNTs are
manipulated by handling their growth substrate, inverted into a
spin-coated polymer layer, and the assembly is cured (usually by
heat). (FIG. 2).
[0055] The CNT's growth substrate may then be removed. The
CNT-growth can be patterned by controlling the patterning of the
thin iron catalyst layer prior to the CNT-growth step. Through
separate control of polymer layer thickness and CNTs length, the
depth of anchoring of the CNTs into the flexible polymer layer can
be controlled. Thus, any pattern of as-grown CNTs, defined by
catalyst pattern may be inverted and anchored into a polymer layer
as required.
[0056] CNTs anchored in PDMS can withstand the shear stress up to
230 dyne/cm.sup.2 and tensile stress up to 64.5 kPa. (Sansom, et
al. 2008 Nanotechnology 19:035302.). The effective adhesion
strength between the CNTs and the flexible polymeric layer depends
on the depth of anchoring of she CNTs inks the polymer layer, the
strength of the bond between CNTs and the polymer layer, and the
fracture toughness of the polymeric layer.
[0057] Anchored CNTs on a flexible polymeric layer can be designed
and prepared such that they have the requisite mechanical strength
ensuring that no CNTs would enter the blood circulation or be left
in the living tissues that may pose harm to the patient. The
biocompatibility of CNTs has been demonstrated, including evidences
that show various types of living ceils (e.g. neuronal cells,
osteoblast cells and fibroblast cells) can be supported by CNTs
(Hu, et al. 2004 Nano Lett 4:507-551; McKenzie, et al. 2004
Biomaterials 25:1309-1317; Webster, et al. 2004 Nanotech 15:48-54;
Gabay, et al. 2005 Physica A 350:611-621; Price, et al. 2005
Biomaterials 24:1877-1887; Elias, et al. 2002 Biomaterials
23:3279-3287; Correa-Duarte, et al. 2004 Nano Lett 4:2233-2236.)
Some studies have been reported that CNTs may present certain
health problems, especially related to lung toxicity, cytotoxicity,
and skin irritation. (Huczko, et al. 2001 Fullerene Sci Tech
9:247-250; Huczko, et al. 2001 Fullerene Sci Tech 9:251-254;
Huczko, et al. 2005 Fullerene Nanotubes Carbon Nanostruct
13:141-145; Lam; et al. 2004 Toxicol Sci 77:126-134; Warheit, et
al. 2004 Toxicol Sci 77:117-125: Muller, et al. 2005 Toxicol App
Pharmacol 207:221-231; Shvedova; et al. 2003 J Toxicol Environ
Health A 66:1909-1926; Monteiro-Rivtere, et al. 2005 Toxicol Lett
155:377-384; Jia, et al. 2005 Environ Sci Technol 39:1378-1383;
Cui, et al. 2005 Toxicol Lett 155:73-85; Tamura, et al. 2004 Key
Eng Mater 254-6:919-922.) Thus, to minimize the risk of side
effects, the minimum mechanical strength of the ACNT on a
CNTs-fitted drug delivery system must be sufficiently higher than
the maximum shear stress induced by the tissue.
[0058] Diffusion is an important mechanism in delivering drugs from
any pharmaceutical system. The release kinetics of a drug delivery
system depends on the initial concentration of the drug and the
surface area of the system. Therefore, to improve the drug delivery
mechanism, it is crucial for a system to have a sufficiently large
surface area. Such a system can essentially be made by fitting an
array of anchored CNT arrays onto a flexible substrate (FIG.
3).
[0059] The following expression compares the surface area of a
CNT-fitted flexible substrate with a bare flexible substrate
without CNT-enhancement.
A CNT - fitted - substrate A bare - substrate = 2 3 .pi. dhWL 3 s 2
WL = 2 3 .pi. dh 3 s 2 ##EQU00002##
Where d is the diameter of individual CNTs, h is the length of
individual CNTs, s is the distance between individual CNT, W and L
are the width and length of the flexible substrate respectively.
Using the following assumed typical values, where d=10 nm, s=100
nm, and h=500 .mu.m, the following result is obtained:
A CNT - fitted - substrate A bare - substrate = 1813.17
##EQU00003##
[0060] This indicates that for a CNT-fitted drug delivery platform,
the mass-transfer surface for delivering drugs is about 1,800 times
greater than that of a non-CNT-fitted platform of similar geometry.
Therefore, there is a substantial increase in drug mass transfer
rate when a drug delivery platform is fitted with CNTs.
[0061] Compared to a simple hollow microneedle, the
surface-to-volume ratio of an anchored CNT bundle is at least one
order of magnitude greater (FIG. 4). This is again crucial for a
local drug delivery platform. This large surface-to-volume ratio
also shows that a high dose of drugs can be placed in a small size
of CNT-fitted drug delivery platform.
[0062] The drug depositing technique for depositing drugs to CNTs
may be any suitable method. A novel approach disclosed here is to
use a very low surface tension liquid (e.g., substantially lower
than 72 dyne/cm (water, 25.degree. Celsius), such as isopropanol,
ethanol and acetone) to place the drugs on the CNTs. Fluid
transport based on wicking through a nano-fibrous material has been
previously studied and characterized. (Zhou, et al. 2006
Nanotechnology 17:4845-4853.) The same phenomenon is employed here
for depositing the drugs in the interstices of the anchored CNT
array from their solutions followed by thorough drying (FIG. 5).
Using this technique, anchored CNTs were coated with both flutax-1
and uranine (sodium salt of fluorescein) (FIGS. 6(a)-(b)).
[0063] The drug depositing technique disclosed herein is a
straightforward and less expensive method to place drugs within an
anchored CNT array. Using this method, drugs can be placed in an
anchored CNT array without having the need to functionalize the
surface of the CNTs. Generally speaking, this method works
preferably for drugs that have one or more aromatic moieties and/or
extended .pi.-bond systems such as atropine and amphetamine, which
help with the creation .pi.-.pi. interactions with CNTs. by
creating strong .pi.-.pi. interactions, the need to functionalize
the CNT beforehand is eliminated. Reduced surface tension liquids
(optionally having a surfactant such as sodium dodecyl sulfate
(SDS) and similar detergents) easily wicked through CNT arrays. For
example, a device can be deposited with the drugs in the
interstices of CNTS using an ethanol solution of the drug followed
by thorough drying to remove the solvent.
[0064] By attaching a flourescent molecule to the drug, e.g.,
taxol, direct photographic observation and monitoring of its
movement into plaque can be accomplished, for example, using
fiber-optic-based technology and calibration methods. (U.S. Pat.
No. 5,116,317 by Carson, et al.; U.S. Pat. No. 4,842,390 by
Sottini, et al.; Tepe. et al. 2007 Touch Briefings
2007-Interventional Cardiology, pp. 61-63; Scheller, et al. 2004
Circulation 110:810-814.) Substantial increases in drug mass
tranfer rate can be achivied when the angioplasty balloon is fitted
with anchored CNTs, as disclosed herein.
[0065] In one embodiment, flutax-1 (i.e., a conjugated paclitaxel
and fluourecein) is used. (Diaz, et al. 2005 J Biol Chem
280:3928-3937; Creel, et al. 2000 Circ Res 86:876-884; Lovich, et
al. 2001 J Pharm Sci 90:1324-1335.) Using flutax-1 allows
measurement of pclitaxel delivery to an arterial-plaque, optically
and directly. This eliminates the need for currently popular
high-performance liquid chromotography (HPLC) approach that would
need biopsy, or for .sup.3H radio-labeled paclitaxel measurement.
(Creel, et al. 2000 Circ Res 86:879-884.) Fabrication of this
device is translatable to industrially scalable processes like toll
-to-toll manufacturing for combining CNTs on substrates and plymer
layers that allows for very low-cost production.
##STR00001##
[0066] The studies disclosed herein include micromechanics and
nano-dynamics relevant to insertion mechanism of nanostructures
into model tissues and directly-on-target interstitial
mass-transfer phenomena involved with such insertion. The ability
to penetrate the arterial plaque cap and deliver the drugs directly
inside the plaque is clearly an outstanding advantage of a
CNT-fitted angioplasty balloon over the conventional
angioplasty-balloon or stent-fitted angioplasty balloon.
[0067] Other studies disclosed herein relate to the prevention of
the carried drugs from being rubbed-out/washed-away during
procedure/travel to the target location. It is important to note
that, due to the hydrophobic nature of CNTs, aqueous media such as
blood plasma do not easily reach the interior spaces of the CNTs
where the drug. Is deposited. By minimizing drug loss, the risks of
overdosing patients may be reduced considerably, another
outstanding advantage of an anchored CNT-fitted angioplasty over
other drug-delivery devices. Studies and results disclosed herein
provide critical information needed tor fabricating specific-target
drug delivery platforms and for guiding the development of novel
drug delivery systems requiring access to interstital spaces.
[0068] Drug protection experiments were conducted to show that the
CNTs protect the deposited drugs from being washed away by
blood-like liquids. Two types of dyes were used in these
experiments, the fluorescein sodium that represents the hydrophilic
drugs and flutax-1 that represents the hydrophobic drugs. Two types
of specimens were used, a bare sheet of latex and a sheet of latex
with CNTs anchored on one side. Since the CNTs are highly
hydrophobic, both fluorescein sodium and flutax-1 were dissolved in
ethanol so that both dyes could go into the interstices of the
CNTs. The same amounts of both fluorescein sodium and flutax-1 were
put in each specimen and allowed to dry. After the dye dried, each
specimen was placed in 2 mL of DI water (to simulate blood) and the
concentration of the dye was analyzed using spectrophotometer.
[0069] The results showed that the hydrophobicity of CNTs protected
the deposited drugs, which could not be easily washed out by DI
water. On the contrary, the DI water was able to wash out the
fluorescein sodium easily (FIG. 7(a)). Notice the obvious
difference of the recovered amount of fluorescein sodium between
the bare latex samples and the CNT-fitted latex samples. The bare
latex samples lost almost all of their initial concentration of
fluorescein sodium, while the CNT-fitted latex samples retained
about 50% of their initial concentration of the fluorescein
sodium.
[0070] Another set of experiments was done using various
concentrations of fluorescein sodium on both types of specimens
(FIG. 7 (b)). The time to wash out the fluorescein sodium in DI
water for the CNT-fitted latex samples depended on the initial
concentration. Lower initial concentration lead to longer time that
was needed to wash out the fluorescein sodium. This was not
observed with the bare latex samples. The bare latex samples could
not retain the fluorescein sodium from being washed away by DI
water for whatever the initial concentration was.
[0071] Another experiment was performed that targeted possible drug
removal from CNT-fitted drug delivery platform by mechanical (e.g.,
rubbing) actions during its cardiovascular travel. Two types of
specimens were used, a bare sheet of latex and a sheet of latex
with CNTs anchored on one side. On each specimen, approximately 125
.mu.g of flutax-1 was deposited by successively dropping small
volumes of its ethanol solution and allowed to dry. Next, 1 .mu.L
of DI water followed by a microscopic 3.times.1.times.5/128-inch
glass slide was put on each specimen. Finally, with a 500 g weight
on top of it, the microscopic slide was moved back and forth a few
millimeters at about 1 cm/sec for 60 seconds (FIGS. 8(a)-(b)). The
results showed considerably higher flutax-1 removal from the bare
latex compared to CNT-anchored one (FIGS. 9(a)-(b)). After the
test, each of the two specimens was dropped in 2 mL of pure ethanol
to see if they still had flutax-1 left on them. There was
considerably more flutax-1 released (present) from the CNT-anchored
latex compared to the bare one (FIG. 10).
[0072] A CNT-fitted angioplasty balloon was prepared with
vertically aligned CNT arrays and anchored on a polymeric layer. We
started with a 20 mm-long PET angioplasty balloon (Advance
Polymers, Item No. 03002016AA). This balloon was then dipped into a
liquid latex compound such that the entire surface of the balloon
was covered uniformly with a thin layer of latex. Before the latex
layer was cured, a silicon substrate, which has as-grown CNTs on
it, was placed in the inverted position onto the latex layer. After
the latex layer was cured, the silicon substrate was then removed,
leaving the CNTs anchored firmly on the latex layer (FIGS.
11(a)-(b)). The right length and packing density of CNTs that allow
optimized drugs delivery and protection can be selected and used
according to specific requirements of a particular application.
[0073] For continuous monitoring of angioplasty drug delivery,
fluorescent-conjugated versions of the drugs intended for delivery,
for example, flutax-1 instead of taxol, can be used. This method
eliminates the need for potentially hazardous administration of a
fluorescent dye intravenously through a central venous line.
Illuminating can be done with a light source to accommodate the
excitation of delivered fluorescent-conjugated drug. A digital
camera can be used to record the imagery of emitted light. (Detter,
et al. 2007 Circulation 116:1007-1014; Hattori, et al. 2009 Circ
Cardiovasc Imaging 2:277-278; Hosono, et al. 2010 Interact
CardioVasc Thorac Surg 10:476-477; Tanaka, et al. 2009 J Thorac
Cardiovasc Surg 138:133-140; Waseda, et al. 2009 JACC
Cardiovascular Imaging 2:604-612.)
[0074] The following studies apply Fick's second law of diffusion
in a semi-infinite medium, x.gtoreq.0. The concentration at the
applicator (angioplasty balloon, etc.) interface is assumed
constant in the first version, diminishing in the second. The first
refers to the initial time-span when applicator is still touching
the plaque, the second refers to post applicator removal.
One-Dimensional Semi-Infinite Slab Model of Solid-Solid Diffusion,
Version I
[0075] This model assumes constant concentration at the applicator
(angio balloon, etc) interface, (Perry, et al. 1963 Chemical
Engineers' Handbook, 4.sup.th edition, McGraw-Hill, Now York.)
.differential. C ( x , t ) .differential. t = D .differential. 2 C
( x , t ) .differential. x 2 Fick ' s second law of diffusion in a
semi - infinite medium , x .gtoreq. 0. ( 1 ) ##EQU00004##
Simplified Initial and Boundary Conditions
[0076] C(x,0)=0 initial condition (2)
c(0,t)=C.sub.0 boundary condition (3)
Simplified Analytical Solution
[0077] Laplace transform with respect to t yields,
.intg. 0 .infin. e - st .differential. C ( x , t ) .differential. t
t = D .intg. 0 .infin. e - st .differential. 2 C ( x , t )
.differential. x 2 t 2 F x 2 - s D F = 0 because of the initial
condition ( 4 ) F ( x , s ) = ( c 0 / s ) e - sx / D , F ( 0 , s )
= c 0 / s because of the boundary condition ( 5 ) ##EQU00005##
[0078] Reverse transform yields,
C ( x , t ) = C 0 [ 1 - 2 .pi. .intg. 0 x / ( 2 Dt ) e - u 2 u ]
for the concentration peak onwards ( 6 ) ##EQU00006##
One-Dimensional Semi-Infinite Slab Model of Solid-solid Diffusion,
Version II
[0079] This model assumes diminishing concentration at the
applicator (angio balloon, etc) interface. (Mehrer 2007 Diffusion
in Solids: Fundamentals, Methods, Materials, Diffusion-Controlled
Processes, Springer-Verlag, Berlin.)
.differential. C ( x , t ) .differential. t = D .differential. 2 C
( x , t ) .differential. x 2 Fick ' s second law of diffusion in a
semi - infinite medium , x .gtoreq. 0. ( 7 ) ##EQU00007##
Simplified Initial and Boundary Conditions
[0080] C(x,0)=M.delta.(x) initial condition (8)
where M is the number of diffusing particles per unit area and O(x)
the Dirac delta function.
.differential. C ( 0 , t ) .differential. x = 0 boundary condition
( 9 ) ##EQU00008##
[0081] The analytical solution is the following Gaussian
equation
C ( x , t ) = M .pi. Dt exp ( - x 2 4 Dt ) which at t = 0 reduces
to C ( x , 0 ) = M .delta. ( x ) ( 10 ) ##EQU00009##
[0082] The diffusion of drugs from coated anchored CNT arrays and
the pressure driven mass transfer through such structures was
experimented with diffusion of flutax-1 from an anchored CNT array
in agar gels and water-containing protein/lipid/cholesterol
structures (including swine and chicken skin). The results
indicated that flutax-1 molecules did not move appreciably in DI
water, but do so in agar (in a matter of minutes) or
gelatin-water-containing matrices. The results with agar gels are
presented below (FIGS. 12(a)-14(b)), and fit reasonably with the
simple theoretical model. The diffusion from the anchored CNT array
was compared with simple diffusion from a thin strip of dried
flutax on a bare polymeric sheet (the geometries of both cases
examined are illustrated in FIGS. 12(a)-(b).
[0083] The diffusion process was modeled with Fick's second law,
assuming one-dimensional diffusion and neglecting the melting time
of the dried flutax in comparison with the diffusion time within
the agar (which is supported by experimental data). Schematic
description of the geometry for both examined cases is presented in
FIGS. 15(b) and 15(c).
[0084] The model for the case of thin layer of flutax (L.sub.1/L1)
consists of one-dimensional diffusion within the agar gel
.differential. C ( x , t ) .differential. t = D .differential. 2 C
( x , t ) .differential. x 2 , ##EQU00010##
and constant concentration C(x=0, t) C.sub.0 and no flux .delta.C
(x=0, t)/ox=0 boundary conditions at x=0 and x.fwdarw..infin.,
respectively. A classical solution for this problem exists in the
literature, and is defined as
C ( x , t ) = C 0 [ 1 - erf ( x 2 Dt ) ] . ##EQU00011##
[0085] For the case of flutax positioned within a CNT array (FIG.
15c) L.sub.2 cannot be neglected in comparison with L, and two
regions of diffusion are modeled, the agar region and the CNT
region. Since the entire length of the CNT array is coated with
flutax at t=0, the initial condition is now
C ( x , t = 0 ) = { 0 , x > L 2 C 0 , x < L 2 ,
##EQU00012##
[0086] Utilizing the one-dimensional case, the reduction in
available area for the diffusion process within the CNT region can
be modeled by reducing the value of the diffusion coefficient
proportionally to the reduction in surface area and the initial
concentration C.sub.0 can be estimated from the amount of flutax
and the available void space within the CNT array.
[0087] FIG. 15(a) presents the numerical solution of the models and
the experimental data for thin flutax layer (squares and smooth
line) and flutax within CNT array (triangles and dashed line). The
general trends of the diffusion are predicted reasonably. The main
effect of the CNT array is to change the initial conditions of the
diffusion and thus the differences between both cases are expected
to decrease as t increase, which is indeed observed in experimental
data in accordance with the model.
EXAMPLES
Experimental Methods
[0088] CNT Arrays Growth.
[0089] The vertically aligned multi-walled carbon nanotube arrays
used in this study were grown using thermal chemical vapor
deposition on silicon wafer substrates. These wafers were coated
with 10 nm aluminum oxide buffer layer and 1 nm iron catalyst layer
using electron beam evaporator (Temescal BJD 1800) and diced into
1.times.1 cm samples. The growth itself was performed in a 1-inch
diameter quartz tube furnace (Lindberg/BlueM Single Zone Tube
Furnace) under the 490 standard cubic centimeters per minute (seem)
ethylene gas (Matheson 99.999%) and 210 seem hydrogen gas (Airgas
99.999%) at a temperature of 750.degree. C. and a pressure of 600
torr. The flow rate and pressure of the gases was maintained by an
electronic mass flow controller (MKS .pi.MFC) and a pressure
controller (MKS .pi.PC). The overall growth quality, including the
length of the array, was characterized under scanning electron
microscope (ZEISS LEO 1550VP).
[0090] Anchored CNT on Flat Surface.
[0091] A thin layer of uncured polymer (e.g., PDMS) was spin-coated
(SCS G3 spin coater) onto a flat rigid substrate. The thickness of
the polymer layer can be controlled by varying the speed (rpm) and
dwell time of the spinner. As-grown carbon nanotube arrays were
manipulated by handling their growth substrate, inverted and then
inserted into the spin-coated polymer layer. The whole assembly was
subsequently cured, usually by heat at elevated temperature (e.g.,
about 80.degree. Celsius). The CNT's growth substrate could then be
easily removed after the polymer layer is fully cured. By
controlling the polymer layer thickness and CNTs length, the depth
of anchoring of the CNTs into the flexible polymer layer may be
controlled.
[0092] Anchored CNT on Balloon.
[0093] A 20 mm-long PET angioplasty balloon (Advance Polymers
03002016AA) was used as a platform. This balloon was then dipped
into a liquid latex compound so that the entire surface of the
balloon was covered uniformly by a thin layer of latex. Before the
latex layer was cured. CNTs attached to a silicon substrate were
inverted then placed into the latex layer. The whole assembly was
subsequently cured by leaving it in air at room temperature for 24
hours. After the latex layer was cured, the silicon substrate was
then removed, leaving the CNTs anchored firmly on the latex layer
on the balloon.
[0094] Flutax-1 and Uranine Attachment on CNT.
[0095] Two types of dye were used in these experiments: the uranine
(sodium fluorescein, Sigma Aldrich 67884) that represents the
hydrophilic drugs and flutax-1 (Tocris Bioscience 2226) that
represents the hydrophobic drugs. Since the carbon nanotubes are
highly hydrophobic, both fluorescein sodium and flutax-1 were
dissolved in pure ethanol (Sigma Aldrich E7023), so that both dyes
could wick into the interstices of the CNT specimen. The same
amounts of both fluorescein sodium and flutax-1 were placed by
successively dropping small volumes in each specimen and letting
them dry in air at room temperature.
[0096] Diffusion Measurement.
[0097] Diffusion of flutax-1 from an anchored CNT array was
measured in 5% agar gels (Sigma Aldrich 17209) in water. The agar
gels were placed inside a 1450 .mu.m OD.times.860 .mu.m ID glass
capillary tube (Clay Adams 4614). The anchored CNT array was then
placed flush to the tip of the capillary tube. To determine the
diffusion profile of the flutax-1 from the CNT array into the agar
gels, time lapsed photographs were taken by fluorescent microscope
(Nikon Eclipse TE2000-S). The diffusion profile was then determined
from the fluorescence intensity captured in these photographs.
INCORPORATION BY REFERENCE
[0098] References and citations to other documents, such as
patents, patent applications, patent publications, journals, books,
papers, web contents, have been made in this disclosure. All such
documents are hereby incorporated herein by reference in their
entirety for all purposes.
EQUIVALENTS
[0099] The representative examples are intended to help illustrate
the invention, and are not intended to, nor should they be
construed to, limit the scope of the invention. Indeed, various
modifications of the invention and many further embodiments
thereof, in addition to those shown and described herein, will
become apparent to those skilled in the art from the full contents
of this document, including the examples and the references to the
scientific and patent literature included herein. The examples
contain important additional information, exemplification and
guidance that can be adapted to the practice of this invention in
its various embodiments and equivalents thereof.
* * * * *