METHODS OF TREATING ERECTILE DYSFUNCTION

Abstract

Therapies for treating erectile dysfunction using regenerative cells and therapeutic energy treatments are disclosed herein. Said therapeutic energy treatments can be selected from one or more of the following methods: electrical stimulation/electroacupuncture, low-level laser irradiation, and extracorporeal shockwave therapy. The combination treatments described herein are useful for restoring components of the penile anatomy that are associated with erectile dysfunction, in particular, nerves, blood vessels and/or smooth muscle cells.

Claims

1. A method of treating erectile dysfunction comprising the steps of: a) identifying an individual suffering from erectile dysfunction; b) selecting a cell with regenerative potential; c) treating said cell with regenerative potential with agents or methods capable of augmenting said regenerative potential as applies to reparation of biological dysfunctions associated with erectile dysfunction; and, d) administering said cell with regenerative potential to said individual suffering from createrectile function.

2. The method of claim 1, wherein said regenerative cell population comprises stem cells selected from the group consisting of a)embryonic stem cells; b) hematopoietic stem cells; c) mesenchymal stem cells; d) very small embryonic like stem cells; e) inducible pluripotent stem cells; f) bone marrow stem cells; g) amniotic fluid stem cells; h) neuronal stem cells; i) parthenogenically derived stem cells; j) umbilical cord blood stem cells; k) placental stem cells; l) germinal stem cells; m) hair follicle stem cells; n) adipose derived stem cells; o) reprogrammed stem cells; p) peripheral blood derived stem cells; q) peripheral blood mesenchymal stem cells; r) endometrial regenerative cells; s) fallopian tube derived stem cells; and, t) dermal stem cells.

3. The cell population of claim 2, wherein said mesenchymal stem cells are derived from a source selected from the group consisting of: a) bone marrow; b) adipose tissue; c) umbilical cord blood; d) Wharton's Jelly; e) inducible pluripotent generated cells; g) placental tissue; h) peripheral blood mononuclear cells; i) differentiated embryonic stem cells; and, j) differentiated progenitor cells.

4. The method of claim 1, wherein said reparation of biological dysfunctions is selected from the group consisting of: a) Restoration of penile smooth muscle mass and/or function; b) Restoration of penile neural tissue and/or its function; and, c) Restoration of penile blood vessels and/or their function(s).

5. The method of claim 1, wherein said regenerative cells are pre-treated with agents capable of inducing stem cell expansion can be selected from TPO, SCF, IL-1, IL-3, IL-7, flt-3L, G-CSF, GM-CSF, Epo, FGF-1, FGF-2, FGF-4, FGF-20, VEGF, activin-A, IGF, EGF, NGF, LIF, PDGF, and a member of the bone morphogenic protein family.

6. The method of claim 1, wherein said methods capable of augmenting said regenerative potential are selecting from the group consisting of: a) Electrical stimulation; b) Shockwave therapy; and, c) Low-level laser irradiation.

7. The method of claim 1, wherein said regenerative cells are administered intracavernosally to said individual in need thereof.

8. A method of treating erectile dysfunction comprising the steps of: a) selecting a cell with regenerative potential; b) administering said cell with regenerative potential to an individual in need of improved erectile function; and, c) administering a therapy to said patient selected from the group consisting of: a) electroacupuncture; b) low-level laser irradiation; and/or, c) shockwave therapy.

9. The method of claim 1, wherein said regenerative cell population comprises stem cells selected from the group consisting of: a) embryonic stem cells; b) hematopoietic stem cells; c) mesenchymal stem cells; d) very small embryonic like stem cells; e) inducible pluripotent stem cells; f) bone marrow stem cells; g) amniotic fluid stem cells; h) neuronal stem cells; i) parthenogenically derived stem cells; j) umbilical cord blood stem cells; k) placental stem cells; l) germinal stem cells; m) hair follicle stem cells; n) adipose derived stem cells; o) reprogrammed stem cells; p) peripheral blood derived stem cells; q) peripheral blood mesenchymal stem cells; r) endometrial regenerative cells; s) fallopian tube derived stem cells; and, t) dermal stem cells.

10. The cell population of claim 2, wherein said mesenchymal stem cells are derived from a source selected from the group consisting of: a) bone marrow; b) adipose tissue; c) umbilical cord blood; d) Wharton's Jelly; e) inducible pluripotent generated cells; g) placental tissue; h) peripheral blood mononuclear cells; i) differentiated embryonic stem cells; and, j) differentiated progenitor cells.

11. The method of claim 1, wherein said regenerative potential refers to the ability of said regenerative cell to repair biological dysfunctions selected from the group consisting of: a) Restoration of penile smooth muscle mass and/or function; b) Restoration of penile neural tissue and/or its function; and, c) Restoration of penile blood vessels and/or their function(s).

12. The method of claim 1, wherein said regenerative cells are treated with agents capable of inducing stem cell expansion can be selected from TPO, SCF, IL-1, IL-3, IL-7, flt-3L, G-CSF, GM-CSF, Epo, FGF-1, FGF-2, FGF-4, FGF-20, VEGF, activin-A, IGF, EGF, NGF, LIF, PDGF, and a member of the bone morphogenic protein family.

13. The method of claim 1, wherein said regenerative cells are administered intracavernosally to said individual in need thereof.

Description

DETAILED DESCRIPTION OF THE EMBODIMENTS

Definitions

[0036] In the context of the present invention, the term regenerative cell refers to a mammalian stem cell, progenitor cell, or differentiated cell. As used herein, embryonic, fetal/placental and adult-derived cellular populations are included in the definition of regenerative cells. In the context of the present invention, regenerative cells may include: embryonic stem cells, cord blood stem cells, placental stem cells, bone marrow stem cells, amniotic fluid stem cells, neuronal stem cells, circulating peripheral blood stem cells, mesenchymal stem cells, germinal stem cells, adipose tissue derived stem cells, exfoliated tooth derived stem cells, hair follicle stem cells, dermal stem cells, parthenogenically derived stem cells, reprogrammed stem cells and side population stem cells and the like.

[0037] As used herein, stem cell refers to a pluripotent cell capable of differentiating into numerous cellular lineages.

[0038] As used herein, the term progenitor cell refers to a lineage-committed cell that may have undergone various stages of differentiation toward a tissue-restricted cell type. The term differentiated cell used in the context of the present invention refers to a tissue-restricted cell.

[0039] As used herein, the term mesenchymal stem cell refers to a multipotent stem cell that can differentiate into a variety of cell types, including: osteoblasts (bone cells), chondrocytes (cartilage cells), and adipocytes (fat cells).

[0040] As used herein, therapeutic energy treatment refers to any light-, acoustic-, electrical-, or mechanical means of stimulating cells or tissue through means either applied externally to cells (i.e. regenerative cells) or directly to the tissue (specifically, the penis in the context of the present invention) for therapeutic purposes.

[0041] As used herein, low level laser irradiation refers to photobiomodulation or phototherapy that may be either stimulatory or inhibitory to cellular functions.

[0042] As used herein, biological dysfunctions used in the context of erectile dysfunction refers to the deficiencies or abnormalities in cellular numbers or functions that underlie the inability of an individual to attain an erection. Said biological dysfunctions are diagnosed in the art using established medical tests and diagnostic criteria. These include but are not limited to the following tests: Doppler Ultrasound, dynamic infusion cavernosometry & cevernosography, tests of penile nerve function including but not limited to the bulbocavernosus reflex test, nocturnal penile tumescence testing, penile biothesiometry, and corpus cavernosometry. Examples of biological dysfunctions present in ED include arteriogenic changes in blood vessels and loss of smooth muscle mass.

[0043] As used herein, a therapeutically effective amount and therapeutically sufficient amount and like terms refer to an amount of an agent sufficient to ameliorate at least one symptom, behavior or event, associated with a pathological, abnormal or otherwise undesirable condition, e.g., erectile dysfunction, or an amount sufficient to prevent or lessen the probability that such a condition will occur or re-occur, or an amount sufficient to delay worsening of such a condition. In one embodiment, the term therapeutically effective amount and like terms are used to refer to the frequencies and concentrations at which regenerative cells are administered for treating erectile dysfunction.

[0044] The present invention comprises a combination therapy involving regenerative cells that and therapeutic energy treatments that are administered to a mammal in need of treatment for erectile dysfunction. Said therapeutic energy treatments include electrical stimulation/electroacupuncture, low level laser therapy, and shock wave therapy. Said therapeutic energy treatments can be administered to said regenerative cells in vitro, and said regenerative cells are subsequently administered to an individual in need of treatment for ED. Said therapeutic energy treatments may also be administered directly to a person in need of treatment for ED. In preferred embodiments of the present invention, said therapeutic energy treatments are administered directly into the penis of an individual with ED prior to or subsequent to administration of said regenerative cells to said individual in need thereof. In the context of the present invention, said regenerative cells are utilized as means for regenerating or repairing nerves, blood vessels and/or smooth muscle cells in order to correct biological dysfunctions associated with ED.

[0045] In certain aspects, the regenerative cells can be selected either alone or in combination from a group that includes: stem cells, committed progenitor cells, and differentiated cells. The stem cells can be selected from a group that includes: embryonic stem cells, cord blood stem cells, placental stem cells, bone marrow stem cells, amniotic fluid stem cells, neuronal stem cells, circulating peripheral blood stem cells, mesenchymal stem cells, germinal stem cells, adipose tissue derived stem cells, exfoliated teeth derived stem cells, hair follicle stem cells, dermal stem cells, parthenogenically derived stem cells, reprogrammed stem cells and side population stem cells and the like.

[0046] In one aspect of the invention, an individual in need of treatment for erectile dysfunction is administered one or several doses of the abovementioned cell types at a therapeutically sufficient concentration and/or frequency.

[0047] In certain aspects of the present invention, the embryonic stem cells can be totipotent, and can express one or more antigens selected from a group that includes: stage-specific embryonic antigens (SSEA) 3, SSEA 4, Tra-1-60 and Tra-1-81, Oct-3/4, Cripto, gastrin-releasing peptide (GRP) receptor, podocalyxin-like protein (PODXL), Rex-1, GCTM-2, Nanog, and human telomerase reverse transcriptase (hTERT) and the like.

[0048] In certain aspects of the present invention, the cord blood stem cells can be multipotent and capable of differentiating into endothelial, smooth muscle, and neuronal cells. The cord blood stem cells can be identified based on expression of one or more antigens selected from a group that includes: SSEA-3, SSEA-4, CD9, CD34, c-kit, OCT-4, Nanog, and CXCR-4 and the like. Further, the cord blood stem cells selected may not express one or more markers selected from a group that includes: CD3, CD34, CD45, and CD11b and the like.

[0049] In certain aspects of the present invention, the placental stem cells can be isolated from the placental structure, and can be identified based on expression of one or more antigens selected from a group that includes: Oct-4, Rex-1, CD9, CD13, CD29, CD44, CD166, CD90, CD105, SH-3, SH-4, TRA-1-60, TRA-1-81, SSEA-4 and Sox-2 and the like.

[0050] In certain aspects of the present invention, the bone marrow stem cells can be bone marrow mononuclear cells, and can be selected based on the ability to differentiate into one or more of the following cell types: endothelial cells, smooth muscle cells, and neuronal cells. The bone marrow stem cells can be selected based on expression of one or more of the following antigens: CD34, c-kit, flk-1, Stro-1, CD105, CD73, CD31, CD146, vascular endothelial-cadherin, CD133 and CXCR-4. Further, the bone marrow stem cells can be enriched for expression of CD133.

[0051] In certain aspects of the present invention, the amniotic fluid stem cells can be isolated by introduction of a fluid extraction means into the amniotic cavity under ultrasound guidance. The amniotic fluid stem cells can be selected based on expression of one or more of the following antigens: SSEA3, SSEA4, Tra-1-60, Tra-1-81, Tra-2-54, HLA class I, CD13, CD44, CD49b, CD105, Oct-4, Rex-1, DAZL and Runx-1. Further, the amniotic fluid stem cells can be selected based on lack of expression of one or more of the following antigens: CD34, CD45, and HLA Class II.

[0052] In certain aspects of the present invention, the neuronal stem cells can be selected based on expression of one or more of the following antigens: RC-2, 3CB2, BLB, Sox-2hh, GLAST, Pax 6, nestin, Muashi-1, NCAM, A2B5 and prominin.

[0053] In certain aspects of the present invention, the circulating peripheral blood stem cells can be characterized by ability to proliferate in vitro for a period of over 3 months. Further, the circulating peripheral blood stem cells can be characterized by expression of CD34, CXCR4, CD117, CD113, and c-met. Further, the circulating peripheral blood stem cells may lack substantial expression of differentiation associated markers, such as, for example CD2, CD3, CD4, CD11, CD11a, Mac-1, CD14, CD16, CD19, CD24, CD33, CD36, CD38, CD45, CD56, CD64, CD68, CD86, CD66b, and HLA-DR and the like.

[0054] In certain aspects of the present invention, the mesenchymal stem cells express one or more of the following markers: STRO-1, CD105, CD54, CD106, HLA-I markers, vimentin, ASMA, collagen-1, fibronectin, LFA-3, ICAM-1, PECAM-1, P-selectin, L-selectin, CD49b/CD29, CD49c/CD29, CD49d/CD29, CD61, CD18, CD29, thrombomodulin, telomerase, CD10, CD13, STRO-2, VCAM-1, CD146, and THY-1. Further, the mesenchymal stem cells may not express substantial levels of HLA-DR, CD117, and CD45. In certain aspects, the mesenchymal stem cells can be derived from a group selected of: bone marrow, adipose tissue, umbilical cord blood, placental tissue, peripheral blood mononuclear cells, differentiated embryonic stem cells, and differentiated progenitor cells.

[0055] In certain aspects of the present invention, the germinal stem cells express markers selected from a group that includes: Oct4, Nanog, Dppa5 Rbm, cyclin A2, Tex18, Stra8, Daz1, beta1- and alpha6-integrins, Vasa, Fragilis, Nobox, c-Kit, Sca-1 and Rex1 and the like.

[0056] In certain aspects of the present invention, the adipose tissue derived stem cells express markers selected from a group that includes: CD13, CD29, CD44, CD63, CD73, CD90, CD166, Aldehyde dehydrogenase (ALDH), and ABCG2 and the like. The adipose tissue derived stem cells can be a population of purified mononuclear cells extracted from adipose tissue capable of proliferating in culture for more than 1 month.

[0057] In certain aspects of the present invention, the exfoliated teeth derived stem cells express markers selected from a group that includes: STRO-1, CD146 (MUC18), alkaline phosphatase, MEPE, and bFGF and the like.

[0058] In certain aspects of the present invention, the hair follicle stem cells express markers selected from a group that includes: cytokeratin 15, Nanog, and Oct-4 and the like, and can be capable of proliferating in culture for a period of at least one month. The hair follicle stem cells may secrete one or more of the following proteins when grown in culture: basic fibroblast growth factor (bFGF), endothelin-1 (ET-1) and stem cell factor (SCF).

[0059] In certain aspects of the present invention, the dermal stem cells express markers selected from a group that includes: CD44, CD13, CD29, CD90, and CD105 and the like, and can be capable of proliferating in culture for a period of at least one month.

[0060] In certain aspects of the present invention, the parthenogenically derived stem cells can be generated by addition of a calcium flux inducing agent to activate an oocyte followed by enrichment of cells expressing markers selected from a group that includes SSEA-4, TRA 1-60 and TRA 1-81 and the like.

[0061] In certain aspects of the present invention, the progenitor cells can be selected from a group that includes: endothelial progenitor cells, neuronal progenitor cells, and hematopoietic progenitor cells and the like.

[0062] In certain aspects, the committed endothelial progenitor cells express markers selected from a group that includes: CD31, CD34, AC133, CD146 and flk1 and the like.

[0063] In certain aspects of the present invention, the committed hematopoietic cells can be purified from the bone marrow, or from peripheral blood, such as from peripheral blood of a patient whose committed hematopoietic progenitor cells can be mobilized by administration of a mobilizing agent or therapy. The mobilizing agent can be selected from a group that includes: G-CSF, M-CSF, GM-CSF, 5-FU, IL-1, IL-3, kit-L, VEGF, Flt-3 ligand, PDGF, EGF, FGF-1, FGF-2, TPO, IL-11, IGF-1, MGDF, NGF, HMG CoA-reductase inhibitors and small molecule antagonists of SDF-1 and the like. Further, the mobilization therapy can be selected from a group that includes: exercise, hyperbaric oxygen, autohemotherapy by ex vivo ozonation of peripheral blood, and induction of SDF-1 secretion outside of the bone marrow.

[0064] In certain aspects of the present invention, the committed hematopoietic progenitor cells can express the marker CD133 or CD34.

[0065] In specific embodiments of the invention, the regenerative cells are expanded in culture using one or more cytokines, chemokines and/or growth factors prior to administration to an individual in need thereof. The agent capable of inducing stem cell expansion can be selected from TPO, SCF, IL-1, IL-3, IL-7, flt-3L, G-CSF, GM-CSF, Epo, FGF-1, FGF-2, FGF-4, FGF-20, VEGF, activin-A, IGF, EGF, NGF, LIF, PDGF, and a member of the bone morphogenic protein family. The agent capable of inducing stem cell differentiation can be selected from HGF, BDNF, VEGF, FGF1, FGF2, FGF4, and FGF 20.

[0066] For use in the present invention, said regenerative cells may be administered to an individual in need thereof via one or more of the following routes of administration: intravenously, intramuscularly, intraperitoneally, transdermally, or by any parenteral route. In a preferred embodiment of the present invention, said regenerative cells are administered intracavernosally.

[0067] Furthermore, conditions promoting certain type of cellular proliferation or differentiation can be used during the culture of said regenerative cells. These conditions include but are not limited to, alteration in temperature, alternation in oxygen/carbon dioxide content, alternations in turbidity of said media, or exposure to small molecules modifiers of cell cultures such as nutrients, inhibitors of certain enzymes, stimulators of certain enzymes, inhibitors of histone deacetylase activity such as valproic acid (Bug, et al., 2005, Cancer Res 65:2537-2541), trichostatin-A (Young, et al., 2004, Cytotherapy 6:328-336), trapoxin A (Kijima, et al., 1993, J Biol Chem 268:22429-22435), or Depsipeptide (Gagnon, et al., 2003, Anticancer Drugs 14:193-202; Fujieda, et al., 2005, Int J Oncol 27:743-748), inhibitors of DNA methyltransferase activity such as 5-azacytidine, inhibitors of the enzyme GSK-3 (Trowbridge, et al., 2006, Nat Med 12:89-98, and the like.

[0068] For use in the present invention, said regenerative cells may be administered to an individual in need thereof via one or more of the following routes of administration: intravenously, intramuscularly, intraperitoneally, transdermally, or by any parenteral route. In a preferred embodiment of the present invention, said regenerative cells are administered intracavernosally (i.e. directly into the corpus cavernosa of the penis).

[0069] In a preferred embodiment of the present invention, said regenerative cells are treated with a therapeutic energy treatment prior to administration to an individual afflicted with erectile dysfunction. Said therapeutic energy treatment may be administered to regenerative cells in vitro and can be selected from one or more of the following: electrical stimulation of cells, low-level laser irradiation of cells, and/or shockwave therapy of cells. Without being bound by theory, these therapeutic energy treatments can increase the therapeutic efficacy of the cellular product used to treat erectile dysfunction by altering one or more of the following activities of regenerative cells: proliferation, differentiation, survival, cytokine and chemokine production, and migration/homing to specific tissue sites.

[0070] In one embodiment, the current invention is practiced by pre-treating regenerative cells with electrical stimulation to modify their therapeutic potential for treating ED. Methods for electrically stimulating cells ex vivo are known in the art. One method for practicing the disclosed invention is described in [38]. Briefly, regenerative cells can be subjected to electrical stimulation using an electrical stimulator consisting of 16 gold electrodes coated with platinum that fit into 8-well chamber slides. Each well contains an anode and cathode with an interelectrode distance of 10 mm. The electrodes can be connected to an electrical stimulator that generates charge-balanced biphasic current pulses. The cells can then be electrically stimulated in suspension for different durations (for example, between 5 and 15 minutes with electric fields of 200 mV/mm at 1 Hz frequencies and 1 ms pulse width). Control regenerative cells can be subjected to the same procedure but without electrical stimulation. After electrical stimulation, the regenerative cells can be cultured in media prior to administration to an individual in need thereof.

[0071] Other devices/apparatus for delivering said electrical stimulation are applicable to the present invention. In the current invention, a therapeutically effective amount of electrical stimulation can be delivered for a period of time sufficient to evoke desirable gene- and protein-expression patterns in said regenerative cells including but not limited to the following: a) Expression of angiogenesis-related genes or proteins; b) Expression of genes or proteins associated with smooth muscle cells or their precursors; and/or, c) Expression of genes or proteins associated with nerve cells or their precursors. Using techniques established in the art to identify one or more of the aforementioned categories of genes and/or proteins, the regenerative potential of electrically-stimulated cells can be compared to that of control cells (non-electrically stimulated) and the suitability of the former for therapeutic purposes (i.e. for treated an individual with ED) can be screened.

[0072] In another embodiment of this invention, regenerative cells can be treated with low-level laser irradiation in vitro to alter their therapeutic potential for the treatment of ED. Techniques for delivering low-level laser irradiation are known in the art. In one embodiment, Low reactive level laser therapy (LLLT) will be delivered directly to cells in tissue culture plates using a laser devices that are commercially available. This invention may also be practiced using different types of laser therapies having variable wavelengths, power outputs/densities, energy densities, types of exposure, and treatment durations. Examples of different types of therapies include single, pulsed, super-pulsed exposure and continuous mode. Various lasers can be utilized including but not limited to diode lasers, HeNe lasers, Er:YAG (Erbium-Doped Yttrium Aluminum Garnet) lasers, and Superpulsed low-level laser therapy (SLLLT). A therapeutically effective amount of laser stimulation can be delivered for a period of time sufficient to evoke desirable gene- and protein-expression patterns in said regenerative cells including but not limited to the following: a) Expression of angiogenesis-related genes or proteins; b) Expression of genes or proteins associated with smooth muscle cells or their precursors; and/or, c) Expression of genes or proteins associated with nerve cells or their precursors.

[0073] In another embodiment of this invention, regenerative cells can be treated with electrocorporeal shockwave therapy in vitro to alter their therapeutic potential for the treatment of ED. This invention may be practiced using methods similar to those described in [39]. Briefly, shockwaves can be applied with a defocused Dermagold 100 device and an OP155 applicator (MTS Medical, Konstanz, Germany). The cells can be stimulated in T25 cell culture flasks, in 15 ml or in 50 ml tubes in PBS. Cells will be submerged in a water bath and stimulated with a frequency of 5 Hz, 200 pulses and energy flux densities ranging from 0.03 to 0.19 mJ/mm.sup.2 at a constant pressure level of 1 bar. A therapeutically effective amount of extracorporeal shockwave therapy can be delivered for a period of time sufficient to evoke desirable gene- and protein-expression patterns in said regenerative cells including but not limited to the following: a) Expression of angiogenesis-related genes or proteins; b) Expression of genes or proteins associated with smooth muscle cells or their precursors; and/or, c) Expression of genes or proteins associated with nerve cells or their precursors.

[0074] Also provided is a method of treating erectile dysfunction comprising administering a therapeutically effective amount of regenerative cells capable of inducing one or more biological activities selected from the group that includes: a) inducing regeneration of nervous tissue; b) stimulating smooth muscle cell activity; c) stimulating increased perfusion or angiogenesis; and, also co-administering therapeutic energy treatments to said individual in need thereof. Said therapeutic energy treatments are selected from one or more of the following: electroacupuncture or TENS, low-level laser irradiation, and/or extracorporeal shockwave therapy. Said therapeutic energy treatments can be administered directly to the penis. Electroacupuncture can be administered at a site distant from the penis using techniques known in the art of alternative medicine. While not being bound by theory, said therapeutic energy treatments can be utilized to enhance the regenerative activities of said regenerative cells or to improve their migration to the penis and/or their retention in the penis. In a preferred embodiment, said regenerative cells may be injected directly into the corpus cavernosa and low-level laser irradiation or extracorporeal shockwave therapy are also administered directly to the penis. Alternatively, said regenerative cells may be injected via another route of administration (for example, intravenously) and said therapeutic energy treatments can be administered to the penile tissue to facilitate migration, survival or other activities of regenerative cells in the penis.

[0075] Thus, provided herein is a method of treating or preventing the onset of erectile dysfunction in a mammal comprising administering a therapeutically effective amount of cells capable of inducing one or more biological activities selected from the group that includes: a) inhibiting neuronal cell dysfunction, b) inhibiting cavernosal fibrosis, c) inhibiting smooth muscle degeneration, and d) inhibiting biological pathways causative of ischemia and the like. Said biological activities of regenerative cells can be modulated by direct treatment of cells with electrical stimulation, shockwave therapy, or low-level laser irradiation. Alternatively, regenerative cells can be administered to an individual for treating erectile dysfunction either as a monotherapy or in a combination therapy with electroacupuncture, low-level laser irradiation, and/or shockwave therapy administered directly to the subject.

[0076] Also provided are methods of treating or preventing the onset of erectile dysfunction in a mammal comprising administering a therapeutically effective amount of cells capable of inducing one or more biological activities selected from the group that includes: a) inducing regeneration of nervous tissue; b) stimulating smooth muscle cell activity; c) stimulating increased perfusion or angiogenesis and the like.

REFERENCES

[0077] 1. Feldman, H. A., et al., Impotence and its medical and psychosocial correlates: results of the Massachusetts Male Aging Study. J Urol, 1994. 151(1): p. 54-61.

[0078] 2. Clavijo, R. I., M. M. Miner, and J. Rajfer, Erectile Dysfunction and Essential Hypertension: The Same Aging-related Disorder? Rev Urol, 2014. 16(4): p. 167-71.

[0079] 3. Magheli, A. and A. L. Burnett, Erectile dysfunction following prostatectomy: prevention and treatment. Nat Rev Urol, 2009. 6(8): p. 415-27.

[0080] 4. Tal, R., et al., Persistent erectile dysfunction following radical prostatectomy: the association between nerve-sparing status and the prevalence and chronology of venous leak. J Sex Med, 2009. 6(10): p. 2813-9.

[0081] 5. Nolan, M. W., et al., Pudendal nerve and internal pudendal artery damage may contribute to radiation-induced erectile dysfunction. Int J Radiat Oncol Biol Phys, 2015. 91(4): p. 796-806.

[0082] 6. Moore, C. S., et al., Erectile dysfunction, vascular risk, and cognitive performance in late middle age. Psychol Aging, 2014. 29(1): p. 163-72.

[0083] 7. Chaudhary, R. K., et al., Risk factors for erectile dysfunction in patients with cardiovascular disease. J Int Med Res, 2016.

[0084] 8. Binmoammar, T. A., et al., The impact of poor glycaemic control on the prevalence of erectile dysfunction in men with type 2 diabetes mellitus: a systematic review. JRSM Open, 2016. 7(3): p. 2054270415622602.

[0085] 9. Bleustein, C. B., et al., The neuropathy of erectile dysfunction. Int J Impot Res, 2002. 14(6): p. 433-9.

[0086] 10. Hurt, K. J., et al., Cyclic AMP-dependent phosphorylation of neuronal nitric oxide synthase mediates penile erection. Proc Natl Acad Sci USA, 2012. 109(41): p. 16624-9.

[0087] 11. Ferguson, J. E., 3rd and C. C. Carson, 3rd, Phosphodiesterase type 5 inhibitors as a treatment for erectile dysfunction: Current information and new horizons. Arab J Urol, 2013. 11(3): p. 222-9.

[0088] 12. Rendell, M. S., et al., Sildenafil for treatment of erectile dysfunction in men with diabetes: a randomized controlled trial. Sildenafil Diabetes Study Group. JAMA, 1999. 281(5): p. 421-6.

[0089] 13. Martinez-Jabaloyas, J. M., et al., Prognostic factors for response to sildenafil in patients with erectile dysfunction. Eur Urol, 2001. 40(6): p. 641-6; discussion 647.

[0090] 14. Deng, W., et al., Gene and stem cell therapy for erectile dysfunction. Int J Impot Res, 2005. 17 Suppl 1: p. S57-63.

[0091] 15. Okolicsanyi, R. K., et al., Human Mesenchymal Stem Cells Retain Multilineage Differentiation Capacity Including Neural Marker Expression after Extended In Vitro Expansion. PLoS One, 2015. 10(9): p. e0137255.

[0092] 16. Foudah, D., et al., Expression of neural markers by undifferentiated mesenchymal-like stem cells from different sources. J Immunol Res, 2014. 2014: p. 987678.

[0093] 17. Eming, S. A., T. Krieg, and J. M. Davidson, Inflammation in wound repair: molecular and cellular mechanisms. J Invest Dermatol, 2007. 127(3): p. 514-25.

[0094] 18. Luster, A. D., R. Alon, and U. H. von Andrian, Immune cell migration in inflammation: present and future therapeutic targets. Nat Immunol, 2005. 6(12): p. 1182-90.

[0095] 19. Ding, Y., et al., Electro-acupuncture promotes survival, differentiation of the bone marrow mesenchymal stem cells as well as functional recovery in the spinal cord-transected rats. BMC Neurosci, 2009. 10: p. 35.

[0096] 20. Genovese, J. A., et al., Electrostimulation induces cardiomyocyte predifferentiation of fibroblasts. Biochem Biophys Res Commun, 2008. 370(3): p. 450-5.

[0097] 21. Genovese, J. A., et al., Cardiac pre-differentiation of human mesenchymal stem cells by electrostimulation. Front Biosci (Landmark Ed), 2009. 14: p. 2996-3002.

[0098] 22. Pietronave, S., et al., Monophasic and biphasic electrical stimulation induces a precardiac differentiation in progenitor cells isolated from human heart. Stem Cells Dev, 2014. 23(8): p. 888-98.

[0099] 23. Pavesi, A., et al., Electrical conditioning of adipose-derived stem cells in a multi-chamber culture platform. Biotechnol Bioeng, 2014. 111(7): p. 1452-63.

[0100] 24. Kushibiki, T., et al., Low Reactive Level Laser Therapy for Mesenchymal Stromal Cells Therapies. Stem Cells Int, 2015. 2015: p. 974864.

[0101] 25. Stein, A., et al., Low-level laser irradiation promotes proliferation and differentiation of human osteoblasts in vitro. Photomed Laser Surg, 2005. 23(2): p. 161-6.

[0102] 26. Stein, E., et al., Initial effects of low-level laser therapy on growth and differentiation of human osteoblast-like cells. Wien Klin Wochenschr, 2008. 120(3-4): p. 112-7.

[0103] 27. Barboza, C. A., et al., Low-level laser irradiation induces in vitro proliferation of mesenchymal stem cells. Einstein (Sao Paulo), 2014. 12(1): p. 75-81.

[0104] 28. Yu, W., J. O. Naim, and R. J. Lanzafame, The effect of laser irradiation on the release of bFGF from 3T3 fibroblasts. Photochem Photobiol, 1994. 59(2): p. 167-70.

[0105] 29. Shen, C. C., et al., Low-level laser stimulation on adipose-tissue-derived stem cell treatments for focal cerebral ischemia in rats. Evid Based Complement Alternat Med, 2013. 2013: p. 594906.

[0106] 30. Waldinger, M. D., et al., Penile anesthesia in Post SSRI Sexual Dysfunction (PSSD) responds to low-power laser irradiation: a case study and hypothesis about the role of transient receptor potential (TRP) ion channels. Eur J Pharmacol, 2015. 753: p. 263-8.

[0107] 31. Wu, J. Y., et al., Low-power laser irradiation suppresses inflammatory response of human adipose-derived stem cells by modulating intracellular cyclic AMP level and NF-kappaB activity. PLoS One, 2013. 8(1): p. e54067.

[0108] 32. Wu, S., et al., High fluence low-power laser irradiation induces mitochondrial permeability transition mediated by reactive oxygen species. J Cell Physiol, 2009. 218(3): p. 603-11.

[0109] 33. Zhou, F., et al., The TGF-beta1/Smad/CTGF pathway and corpus cavernosum fibrous-muscular alterations in rats with streptozotocin-induced diabetes. J Androl, 2012. 33(4): p. 651-9.

[0110] 34. Liu, J., et al., Evaluation of the effect of different doses of low energy shock wave therapy on the erectile function of streptozotocin (STZ)-induced diabetic rats. Int J Mol Sci, 2013. 14(5): p. 10661-73.

[0111] 35. Olsen, A. B., et al., Can low-intensity extracorporeal shockwave therapy improve erectile dysfunction? A prospective, randomized, double-blind, placebo-controlled study. Scand J Urol, 2015. 49(4): p. 329-33.

[0112] 36. Vardi, Y., et al., Can low-intensity extracorporeal shockwave therapy improve erectile function? A 6-month follow-up pilot study in patients with organic erectile dysfunction. Eur Urol, 2010. 58(2): p. 243-8.

[0113] 37. Gruenwald, I., et al., Shockwave treatment of erectile dysfunction. Ther Adv Urol, 2013. 5(2): p. 95-9.

[0114] 38. Hernandez, D., et al., Electrical Stimulation Promotes Cardiac Differentiation of Human Induced Pluripotent Stem Cells. Stem Cells Int, 2016. 2016: p. 1718041.

[0115] 39. Rohringer, S., et al., Molecular and cellular effects of in vitro shockwave treatment on lymphatic endothelial cells. PLoS One, 2014. 9(12): p. e114806.