Angiogenesis initiation and growth

1. A method for promoting angiogenesis at a site on a patient, the method comprising:

obtaining blood compatible with the patient;

isolating platelet-rich plasma by density fractionation of said blood;

administering said platelet-rich plasma to the site; and

stimulating the site with an electromagnetic field following administration of said platelet-rich plasma to the site, thereby

promoting angiogenesis at the site upon application of said electromagnetic field and said platelet-rich plasma to the site.

2. The method for promoting angiogenesis at a site on a patient as set forth in claim 1 , wherein isolating platelet-rich plasma by density fractionation comprises centrifuging of said blood by:

loading said blood into a tube having a buoy;

spinning said tube so that said buoy defines an interface between platelet-rich plasma and platelet poor plasma contained within said blood; and

collecting said platelet-rich plasma.

3. The method for promoting angiogenesis at a site on a patient as set forth in claim 1 , wherein stimulating the site with the electromagnetic field following the administration of said platelet-rich plasma includes generating in the tissue a capacitively coupled time varying electromagnetic field having a peak amplitude of from about 0.1 to about 100 millivolts per centimeter, and having a frequency from about 1 kHz to about 200 kHz.

4. The method for promoting angiogenesis at the site on a patient as set forth in claim 1 , wherein stimulating the site with an electromagnetic field following the administration of said platelet-rich plasma includes generating a pulsed electromagnetic field having a pulse duration from about 10 to 2000 microseconds.

5. The method for promoting angiogenesis at a site on a patient as set forth in claim 1 , wherein promoting angiogenesis of the site upon application of said electromagnetic field and said platelet-rich plasma comprises:

promoting the production of FGF-2 at the site upon the application of said electromagnetic field; and

upregulating the proliferation of endothelial cells at the site upon application of said electromagnetic field.

6. The method for promoting angiogenesis at a site on a patient as set forth in claim 1 , further comprising administering stem cells to the site, said stem cells selected from the group consisting of adipose stromal cells, stem cells derived from bone marrow aspirate, and combinations thereof.

7. The method for promoting angiogenesis at a site on a patient as set forth in claim 6 , wherein administering stem cells to the site comprises:

performing lipoaspiration on the patient to obtain adipose tissue, said adipose tissue including adipose stem cells and adipocytes;

enzymatically digesting said adipose tissue; and

separating said adipose stem cells from said adipocytes in said enzymatically digested adipose tissue.

8. The method for promoting angiogenesis at a site on a patient as set forth in claim 6 , wherein administering stem cells to the site comprises:

isolating bone marrow aspirate from the patient, said bone marrow aspirate including stem cells;

centrifuging said bone marrow aspirate to concentrate said stem cells;

isolating said concentrated stem cells derived from bone marrow aspirate.

9. The method for promoting angiogenesis at a site on a patient as set forth in claim 1 , wherein administering platelet-rich plasma to the site further comprises administering an isolated angiogenic factor selected from the group consisting of angiogenin, angiopoietin-1, del-1 protein, acidic fibroblast growth factor (aFGF or FGF-1), basic fibroblast growth factor (bFGF or FGF-2), follistatin, granulocyte colony-stimulating factor (G-CSF), hepatocyte growth factor (HGF), interleukin-8 (IL-8), leptin, midkine, placental growth factor, platelet-derived endothelial growth factor (PD-ECGF), platelet-derived growth factor (PDGF), pleiotrophin (PTN), progranulin, proliferin, transforming growth factor alpha (TGF-α), transforming growth factor beta (TGF-β), tumor necrosis factor alpha (TNF-α), vascular endothelial growth factor (VEGF), vascular permeability factor (VPF), and combinations thereof.

10. The method for promoting angiogenesis at a site on a patient as set forth in claim 1 , wherein the site on the patient exhibits one of the following conditions: coronary artery disease, stroke, delayed wound healing, chronic wound, peripheral vascular disease, ischemia, chronic tendonosis, wound or tissue breakdown resulting from radiation treatment, and myocardial infarction.

11. The method for promoting angiogenesis at a site on a patient as set forth in claim 1 , wherein the site on the patient exhibits one of the following conditions: stroke, peripheral vascular disease, ischemia, and wound or tissue breakdown resulting from radiation treatment.

12. A method for promoting angiogenesis at the site of an ulcer on a patient comprising:

obtaining blood compatible with the patient;

isolating platelet-rich plasma by loading said blood into a tube having a buoy, spinning said tube so that said buoy defines an interface between platelet-rich plasma and platelet-poor plasma contained within said blood, and collecting said platelet-rich plasma;

administering said platelet-rich plasma to the site of the ulcer; and

stimulating the site with an electromagnetic field following administration of said platelet-rich plasma to the site, thereby

promoting angiogenesis at the site upon application of said electromagnetic field and said platelet-rich plasma to the site.

13. The method for promoting angiogenesis at a site on a patient as set forth in claim 12 , wherein stimulating the site with the electromagnetic field following the administration of said platelet-rich plasma includes generating in the tissue a capacitively coupled time varying electromagnetic field having a peak amplitude of from about 0.1 to about 100 millivolts per centimeter, and having a frequency from about 1 kHz to about 200 kHz.

14. The method for promoting angiogenesis at the site on a patient as set forth in claim 12 , wherein stimulating the site with an electromagnetic field following the administration of said platelet-rich plasma includes generating a pulsed electromagnetic field having a pulse duration from about 10 to 2000 microseconds.

15. The method for promoting angiogenesis at a site on a patient as set forth in claim 12 , further comprising administering stem cells to the site, wherein said stem cells are selected from the group consisting of adipose stromal cells, stem cells derived from bone marrow aspirate, and combinations thereof.

16. The method for promoting angiogenesis at a site on a patient as set forth in claim 15 , wherein administering stem cells to the site comprises:

isolating bone marrow aspirate from the patient, said bone marrow aspirate including stem cells;

centrifuging said bone marrow aspirate to concentrate said stem cells;

isolating said concentrated stem cells derived from bone marrow aspirate.

17. The method for promoting angiogenesis at a site on a patient as set forth in claim 12 , wherein administering platelet-rich plasma to the site further comprises administering an isolated angiogenic factor selected from the group consisting of angiogenin, angiopoietin-1, del-1 protein, acidic fibroblast growth factor (aFGF or FGF-1), basic fibroblast growth factor (bFGF or FGF-2), follistatin, granulocyte colony-stimulating factor (G-CSF), hepatocyte growth factor (HGF), interleukin-8 (IL-8), leptin, midkine, placental growth factor, platelet-derived endothelial growth factor (PD-ECGF), platelet-derived growth factor (PDGF), pleiotrophin (PTN), progranulin, proliferin, transforming growth factor alpha (TGF-α), transforming growth factor beta (TGF-β), tumor necrosis factor alpha (TNF-α), vascular endothelial growth factor (VEGF), vascular permeability factor (VPF), and combinations thereof.

18. A method for promoting angiogenesis at the site of a tissue defect in a patient, comprising:

obtaining blood compatible with the patient;

isolating platelet-rich plasma by loading said blood with a tube having a buoy, spinning said tube so that said buoy defines an interface between platelet-rich plasma and platelet-poor plasma contained within said blood, and collecting said platelet-rich plasma;

administering said platelet-rich plasma to the site; and

stimulating the site with an electromagnetic field following administration of said platelet-rich plasma to the site, thereby

promoting angiogenesis at the site upon application of said electromagnetic field and said platelet-rich plasma to the site;

wherein said tissue defect is stroke, peripheral vascular disease, ischemia, or wound or tissue breakdown resulting from radiation treatment.

19. The method for promoting angiogenesis at a site on a patient as set forth in claim 18 , wherein stimulating the site with the electromagnetic field following the administration of said platelet-rich plasma includes generating in the tissue a capacitively coupled time varying electromagnetic field having a peak amplitude of from about 0.1 to about 100 millivolts per centimeter, and having a frequency from about 1 kHz to about 200 kHz.

20. The method for promoting angiogenesis at the site on a patient as set forth in claim 18 , wherein stimulating the site with an electromagnetic field following the administration of said platelet-rich plasma includes generating a pulsed electromagnetic field having a pulse duration from about 10 to 2000 microseconds.

21. The method for promoting angiogenesis at a site on a patient as set forth in claim 18 , further comprising administering stem cells to the site, wherein said stem cells are selected from the group consisting of adipose stromal cells, stem cells derived from bone marrow aspirate, and combinations thereof.

22. The method for promoting angiogenesis at a site on a patient as set forth in claim 21 , wherein administering stem cells to the site comprises:

isolating bone marrow aspirate from the patient, said bone marrow aspirate including stem cells;

centrifuging said bone marrow aspirate to concentrate said stem cells;

isolating said concentrated stem cells derived from bone marrow aspirate.

23. The method for promoting angiogenesis at a site on a patient as set forth in claim 18 , wherein administering platelet-rich plasma to the site further comprises administering an isolated angiogenic factor selected from the group consisting of angiogenin, angiopoietin-1, del-1 protein, acidic fibroblast growth factor (aFGF or FGF-1), basic fibroblast growth factor (bFGF or FGF-2), follistatin, granulocyte colony-stimulating factor (GCSF), hepatocyte growth factor (HGF), interleukin-8 (IL-8), leptin, midkine, placental growth factor, platelet-derived endothelial growth factor (PD-ECGF), platelet-derived growth factor (PDGF), pleiotrophin (PTN), progranulin, proliferin, transforming growth factor alpha (TGF-α), transforming growth factor beta (TGF-β), tumor necrosis factor alpha (TNF-α), vascular endothelial growth factor (VEGF), vascular permeability factor (VPF), and combinations thereof.