U.S. patent application number 11/968393 was filed with the patent office on 2008-07-10 for methods and materials for stimulating proliferation of stem cell.
Invention is credited to Sudipta Seal, Kiminobu Sugaya.
Application Number | 20080166412 11/968393 |
Document ID | / |
Family ID | 39589015 |
Filed Date | 2008-07-10 |
United States Patent
Application |
20080166412 |
Kind Code |
A1 |
Sugaya; Kiminobu ; et
al. |
July 10, 2008 |
METHODS AND MATERIALS FOR STIMULATING PROLIFERATION OF STEM
CELL
Abstract
Disclosed herein are methods and materials for influencing
proliferation of stem cells. Specifically exemplified herein are
compositions comprising cerium oxide nanoparticles which can be
used to stimulate proliferation of stem cells under common culture
conditions, or which can be utilized to improve therapeutic
outcomes.
Inventors: |
Sugaya; Kiminobu; (Winter
Park, FL) ; Seal; Sudipta; (Oviedo, FL) |
Correspondence
Address: |
Timothy H. Van Dyke
390 No. Orange Avenue, Suite 2500
Orlando
FL
32801
US
|
Family ID: |
39589015 |
Appl. No.: |
11/968393 |
Filed: |
January 2, 2008 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60883081 |
Jan 2, 2007 |
|
|
|
Current U.S.
Class: |
424/489 ;
424/617; 424/93.7; 435/375; 977/773 |
Current CPC
Class: |
A61K 35/28 20130101;
B82Y 5/00 20130101; A61K 9/51 20130101; Y10S 977/896 20130101; Y10S
977/773 20130101; Y10S 977/915 20130101; C01F 17/206 20200101; A61K
33/24 20130101; A61K 33/00 20130101; A61P 25/28 20180101; B82Y
40/00 20130101; A61K 2300/00 20130101; A61K 2300/00 20130101; A61K
35/28 20130101; A61K 33/00 20130101 |
Class at
Publication: |
424/489 ;
424/93.7; 435/375; 424/617; 977/773 |
International
Class: |
A61K 9/16 20060101
A61K009/16; A61K 35/12 20060101 A61K035/12; A61K 33/24 20060101
A61K033/24; C12N 5/02 20060101 C12N005/02 |
Claims
1. A method of influencing proliferation of stem cells in a body
comprising administering stem cells to a body; and administering a
composition comprising cerium oxide to said body, wherein said stem
cells and said composition are placed in said body such that said
migration influencing factor is brought into contact with at least
one of said stem cells.
2. The method of claim 2, wherein said composition is an admixture
of cerium oxide and a pharmaceutically acceptable carrier.
3. The method of claim 2, wherein said pharmaceutically acceptable
carrier comprises water.
4. The method of claim 1, wherein said composition is administered
via an intravenous, intraarterial, or intramuscular route of
administration.
5. A method of stimulation proliferation of a population of stem
cells comprising contacting said population with cerium oxide
nanoparticles, wherein said contacting occurs in vitro, in vivo, in
situ and/or ex vivo.
6. The method of claim 5, wherein said nanoparticles are of a size
from about 0.5 nm to about 50 nm.
7. The method of claim 6, wherein said nanoparticles are spherical
shaped and about 3-5 nm in size.
8. The method of claim 6 wherein said nanoparticles are spherical
in shape and about 10-20 nm in size.
9. The method of claim 6, wherein said nanoparticles comprise
agglomerates of 3-5 nm sized individual crystals.
10. The method of claim 6, wherein said nanoparticles are spherical
in shape and about 11-15 nm in size.
11. The method of claim 6, wherein said nanoparticles are spherical
in shape and about 7-10 nm in size.
12. The method of claim 6, wherein said nanoparticles are cuboidal
in shape and about 10-20 nm in size.
13. A method of treating or preventing a neurological or corporal
deficit comprising administering a therapeutically effective amount
of cerium oxide nanoparticles.
14. The method of claim 13, wherein said composition is
administered via parenteral administration.
15. The method of claim 13, wherein parenteral administration
comprises intramuscular, intravenous, intracistern, intraventrical,
intraarterial, or local administration.
16. The method of claim 13, wherein said administration comprises,
rectal administration.
17. The method of claim 13, wherein said administration comprises
topical administration.
18. The method of claim 13, wherein said administration comprises
dermal or ocular.
19. The method of claim 13, wherein said composition is
administered via oral administration.
Description
RELATED APPLICATIONS
[0001] This application claims priority to U.S. Ser. No.
60/883,081, filed Jan. 2, 2007, which is incorporated herein by
reference in its entirely.
BACKGROUND OF THE INVENTION
[0002] Patients currently suffering from neurodegenerative
conditions such as Alzheimer's and Parkinson's have limited
treatment options. Conventional drug therapy helps delay or reduce
the symptoms of disease but is unable to restore complete
functionality of the brain or repair damaged tissue. Through stem
cell-based therapies, scientists aim to transplant cells in order
to regenerate damaged tissue and restore proper function. In
addition to addressing neurodegenerative diseases, stem cell based
therapies show much promise in addressing a vast number of other
diseases, injuries and conditions. The discovery of safe and
inexpensive compositions to modulate stem cell proliferation would
be a beneficial advancement in the filed of stem cell-based
therapies and basic research.
DETAILED DESCRIPTION
[0003] Scientists are beginning to realize the potential power of
stem-cell based therapies. It is widely recognized that stem cell
transplantation may lead to treatments and/or cures of baneful
disease states, degeneration and/or injury. The ability to
stimulate the proliferation of stem cells in vivo or in vitro will
be of tremendous benefit. The subject invention is based on the
inventors discovery that certain nanoparticles can affect the
proliferation of stem cells.
[0004] In one embodiment, the subject invention pertains to a
method of improving therapeutic outcomes of implanted stem cells
comprising administering a Cerium Oxide (CeO.sub.2) nanoparticle
composition in conjunction with stem cell implantation. The
administration of the Cerium Oxide composition can be at or
proximate to a target stem cell implantation site in a human or
nonhuman animal. In an alternative embodiment, the environment may
be further modified by provision of influencing factors or by
cleaning the environment of undesired or toxic agents that may
affect administered stem cells in an undesired way. The
administered stem cells may be unmodified or may themselves be
engineered to be biased toward a target differentiation endpoint.
U.S. Patent Publication Nos. 20060134789 and 20060110440 are
incorporated by reference to provide examples of stem cells
engineered for negative and positive differentiation biasing that
are contemplated for use with the methods taught herein.
[0005] In another embodiment, the subject invention involves the
administration of a composition comprising a administering Cerium
Oxide nanoparticle composition to a body and administering stem
cells to said body. The composition is administered at a locus in
said body so as to allow contact with the stem cells. This may be
at the same location, proximate to the location or distal to the
location of where stem cells are administered.
[0006] Stem cells may be administered by injecting one or a
plurality of stem cells with a syringe, inserting the stem cells
with a catheter or surgically implanting the stem cells. In certain
embodiments, the stem cells are administered into a body cavity
fluidly connected to a target tissue. In certain preferred
embodiments, the body cavity is a brain ventricle. In other
embodiments, the stem cells are inserted using a syringe or
catheter, or surgically implanted directly at the target tissue
site. In other embodiments, the stem cells are administered
parenterally. Parenteral administration is historically defined as
administration via a route that bypasses the gastrointestinal
tract. Therefore, technically parenteral administration includes
intraventricular administration, but intraventricular
administration is typically discussed herein as separate from other
modes of administration that may fall under `parenteral
administration`.
[0007] The compositions can be administered alone or in combination
with at least one other agent, such as stabilizing or buffer
compounds, which can be administered in any sterile, biocompatible
pharmaceutical carrier, including, but not limited to, saline,
buffered saline, dextrose, and water. In addition to the critical
components of compositions discussed herein, cells or influencing
factors, the compositions can contain suitable pharmaceutically
acceptable carriers comprising excipients and auxiliaries which
facilitate processing of the active compounds into preparations
which can be used pharmaceutically. Generally compositions can be
administered by any number of routes including, but not limited to,
oral, intravenous, intramuscular, intra-arterial, intramedullary,
intrathecal, intraventricular, transdermal, subcutaneous,
intraperitoneal, intranasal, parenteral, topical, sublingual, or
rectal means. Factors can be administered at the same location as
administered stem cells. Administration of influencing factors and
stem cells can be conducted simultaneously, or one prior to the
other, and at the same or different locations so long as the
relative locations allow for the factors to influence the stem
cells.
[0008] The administration of cerium oxide nanoparticles to stem
cells can be conducted in vivo, ex vivo, in situ, and/or in vitro.
According to a specific embodiment, cerium oxide nanoparticles are
applied to stem cells in vitro. This will aid and facilitate basic
experiments conducted on stem cells that will benefit from
stimulation of the stem cells or to create for the stem cells an
environment where oxidants are minimized. In other embodiments, the
invention is directed to the stem cell proliferating cerium oxide
nanoparticle compositions themselves.
[0009] In other embodiments, the environment of administered stem
cells is modified by administration of a separate population of
stem cells that are engineered to express influencing factors.
Engineered stem cells designed to express influencing factors may
be produced according to previously disclosed methods. See for
example U.S. Patent Publication Nos. 20060134789 and 20060110440.
According to a specific method embodiment, a population of
therapeutic stem cells are administered to a locus of a subject's
body and a population of engineered stem cells is administered at
or proximate to the therapeutic population. Influencing factors are
expressed and released from the engineered stem cells which in turn
influence the therapeutic stem cells for a desired objective.
[0010] Thus, the invention also provides improved methods of
treating health conditions implementing a stem cell strategy. For
example, the methods taught herein may be used to treat or
ameliorate symptoms due to a corporal or neurological (affecting
nervous system), such as a deficit caused by a neurodegenerative
disease, a traumatic injury, a neurotoxic injury, ischemia, a
developmental disorder, a disorder affecting vision, an injury or
disease of the spinal cord, a demyelinating disease, an autoimmune
disease, an infection, an inflammatory disease, or corporal
disease, disorder, injury, trauma, malfunction, degeneration or
loss. See, for example, U.S. Patent Publication No. 20030139410. In
preferred embodiments, implanted stem cells are capable of
proliferating, migrating to an area of tissue damage, and/or
differentiating in a tissue-specific manner and functioning in a
manner that reduces the neurological or corporal deficit. Stem
cells may be administered by injection with a syringe, inserting
cells with a catheter or surgically implanting cells. In certain
embodiments, stem cells are injected with a syringe into a body
cavity that is fluidly-connected to the area of neurological or
corporal deficit. In certain preferred embodiments, the body cavity
is a brain ventricle. In other embodiments, stem cells are inserted
with a catheter into a body cavity that is fluidly-connected to the
area of neurological or corporal deficit. In certain preferred
embodiments, the body cavity is a brain ventricle. In still further
additional embodiments, stem cells are surgically implanted into a
body cavity that is fluidly-connected to the area of neurological
or corporal deficit. In certain preferred embodiments, the body
cavity is a brain ventricle. Stem cells can also alternatively be
inserted using a syringe or catheter or surgically implanted
directly at the site of the neurological or corporal deficit or
systemically (e.g., intravenously).
[0011] The invention provides methods of delivery and
transplantation of stem cells to ameliorate the effects of age,
physical and biological trauma and degenerative disease on the
brain or central nervous system of a human or nonhuman animal, as
well as other tissues such as, for example, retinal tissue. It is
well recognized in the art that transplantation of tissue into the
CNS offers the potential for treatment of neurodegenerative
disorders and CNS damage due to injury. Transplantation of new
cells into the damaged CNS has the potential to repair damaged
circuitries and provide neurotransmitters thereby restoring
neurological function. It is also recognized in the art that
transplantation into other tissue, such as eye tissue, offers the
potential for treatment of degenerative disorders and tissue damage
due to injury. As disclosed herein, the invention provides methods
for methods of modifying a tissue environment to influence
proliferation, migration and differentiation of administered stem
cells when introduced into a target site. In a certain specific
embodiments, target sites may be those appropriate for treatment of
neurological disorders and CNS damage.
[0012] Recipients of administered stem cells can be
immunosuppressed, either through the use of immunosuppressive drugs
such as cyclosporin, or through local immunosuppression strategies
employing locally applied immunosuppressants, but such
immunosuppression need not necessarily be a prerequisite in certain
immunoprivileged tissues such as, for example, brain and eye
tissues. In certain embodiments, the delivery method of the
invention can cause less localized tissue damage to the site of
cell damage or malfunction than existing methods of delivery.
[0013] In certain embodiments, administered stem cells are
autologous in nature, i.e., prepared from the recipient's own
tissue. In such instances, the progeny of stem cells can be
generated from dissociated or isolated tissue and proliferated in
vitro using known methods. In the case of mesenchymal stem cells
(MeSCs), progeny can be generated from MeSCs isolated from, for
example, bone marrow. Upon suitable expansion of cell numbers, the
stem cells of the invention can be harvested and readied for
administration into the recipient's affected tissue.
[0014] Reference to particular buffers, media, reagents, cells,
culture conditions and the like, or to some subclass of same, is
not intended to be limiting, but should be read to include all such
related materials that one of ordinary skill in the art would
recognize as being of interest or value in the particular context
in which that discussion is presented. For example, it is often
possible to substitute one buffer system or culture medium for
another, such that a different but known way is used to achieve the
same goals as those to which the use of a suggested method,
material or composition is directed.
[0015] It is important to an understanding of the present invention
to note that all technical and scientific terms used herein, unless
defined herein, are intended to have the same meaning as commonly
understood by one of ordinary skill in the art. The techniques
employed herein are also those that are known to one of ordinary
skill in the art, unless stated otherwise. For purposes of more
clearly facilitating an understanding the invention as disclosed
and claimed herein, the following definitions are provided.
EXAMPLE 1
Preparation of Cerium Oxide Nanoparticles
[0016] Cerium oxide nanoparticles of a size approximately 2 nm to
approximately 10 nm in diameter, are prepared by a process
including the steps of dissolving approximately 0.5 grams to
approximately 1.0 grams of Ce(NO.sub.3).sub.3.6H.sub.2O in
deionized water to make approximately 10 mls of solution to form a
first solution, followed by dissolving approximately 3 grams to
approximately 4 grams of AOT (surfactant) in approximately 200 ml
of solvent to form a second solution, followed by combining the
first and the second solutions, followed by stirring the combined
solutions for approximately 30 minutes, and drop wise adding
approximately 30% hydrogen peroxide (H.sub.2O.sub.2) until the
stirred combined solution becomes yellow, and subsequently stirring
for approximately 30 minutes to approximately 60 minutes more.
Thus, aqueous reverse micelles (RMs) are surfactant aggregates in
nonpolar solvents that enclose packets of aqueous solution in their
interior. The size of the water droplet can be tuned by varying the
ratio of water to surfactant. RMs used as reaction media in the
production of nanoparticles whose size and shape are controlled by
water and surfactant ratio. The surfactant is stripped off using
simple washing and drying. Size: 3-5 nm.
EXAMPLE 2
[0017] The synthesis of ceria was carried out by direct synthesis
route from the cerium precursor (Ce(NO.sub.3).sub.3.6H.sub.2O)
dissolved in water. Oxidation of cerium nitrate was carried out
using hydrogen peroxide (H.sub.2O.sub.2). It is a very simple one
step process yielding ceria nanoparticles dispersed in water (a
very useful delivery media for biological studies). The process
leads to formation of cerium oxide nanoparticles of 15-20 nm
particle size (agglomerates) consisting 3-5 nm spherical
crystals.
EXAMPLE 3
[0018] Particles may be obtained via heating of the particles in
produced in Example 1 above, anywhere between 100-400 C.
EXAMPLE 4
[0019] Cerium oxide particles are obtained from commercially
available sources. Particles are dried from the slurry, washed in
water and redispersed in the water at a certain pH. Without being
bound to any particular theory, it is believed that the variation
in the stem cell proliferation, though not yet shown to be
statistically significant, appears to be attributable to difference
in the Ce+3/Ce+4 ratio in the samples.
EXAMPLE 5
Stimulation of Stem Cell Proliferation
[0020] Stem cell growth and division rates can be increased by
lowering the oxidative stress on the cells. We propose using this
method to increase the proliferation rates of stem cells under
common culture conditions. Specifically, we can improve stem cell
proliferation rates by as much as 50%. Furthermore, we propose that
this method can be used to enable the growth of stem cells for
emergency tissue repair as well as for therapeutic use. This
therapy can be used in conjunction with currently available drugs
and biologics with little concern over negative interactions and
helps eliminate the possibility for graft-verse-host disease. Using
five different samples of cerium oxide, CeO2, nanoparticles, size
and shape, 3-5 nm spherical, 11-15 nm agglomerates of 3-5 nm
individual crystals, 10-20 nm spherical, 7-10 nm spherical, and
10-20 nm cuboidal, the inventors found a good working concentration
by making serial dilutions of each sample, then applied each for a
plated dish of rat mesenchymal stem cells, also known as adult stem
cells. Although a wide range of concentrations can be used, the
inventors found that 10 nM appeared to be an optimal concentration.
When rat and human mesenchymal stem cells were plated at known
densities, treated with one of the CeO2 samples, allowed to grow
for periods of 24, 48, and 72 hours, then performed a cell count
for each of the samples and the control untreated cells, the rate
of cell proliferation had a marked increase. Though all samples
showed an influence on proliferation, Sample 3, which is composed
of CeO2 nanoparticles 10-20 nm in size and spherically shaped and
showed highest increase, increased mesenchymal stem cell
proliferation rates by as much as 50%.
[0021] While a number of embodiments of the present invention have
been shown and described herein in the present context, such
embodiments are provided by way of example only, and not of
limitation. Numerous variations, changes and substitutions will
occur to those of skilled in the art without materially departing
from the invention herein. It should be borne in mind that all
patents, patent applications, patent publications, technical
publications, scientific publications, and other references
referenced herein are hereby incorporated by reference in this
application, in their entirety to the extent not inconsistent with
the teachings herein.
* * * * *