Methods and compositions to facilitate repair of avascular tissue

Abstract

Compositions and methods are provided for repairing damaged avascular zones, including intervertebral disc, in a patient in need thereof.

What is claimed is: 
     
         1 . A method for treating an avascular zone in a patient in need thereof comprising:
 culturing harvested nucleated cells from the patient in need thereof in a culture medium under a selective pressure of about 1% to about 10% oxygen and a pH of about 6.6 to about 7.0;   selecting viable, undifferentiated mesenchymal stem cells capable of growth in the culture medium under the selective pressure of about 1% to about 10% oxygen and a pH of about 6.6 to about 7.0; and   providing the selected, viable, undifferentiated mesenchymal stem cells for implantation in the avascular zone.   
     
     
         2 . The method of  claim 1 , wherein the selective pressure is about 3 to about 7% oxygen. 
     
     
         3 . The method of  claim 1 , wherein the selective pressure further includes from about 2 to about 10% carbon dioxide. 
     
     
         4 . The method of  claim 1 , wherein the selective pressure further includes a pH of about 6.7 to about 6.9. 
     
     
         5 . The method of  claim 1 , wherein the culture medium is supplemented with about 10 to about 20% platelet cell lysate. 
     
     
         6 . The method of  claim 1 , further comprising:
 harvesting platelets from the patient in need thereof; and   implanting the platelets prior to, during, or after implanting the selected, viable, undifferentiated mesenchymal stem cells;   wherein the implanting of the platelets and the selected, viable, undifferentiated mesenchymal stem cells is in the avascular zone.   
     
     
         7 . The method of  claim 6 , wherein the platelets are treated with thrombin and calcium chloride 1-7 days prior to implanting. 
     
     
         8 . The method of  claim 7 , wherein the amount of thrombin is about 28.56 U/ml and the amount of calcium chloride is about 2.86 mg/ml. 
     
     
         9 . The method of  claim 6 , wherein the platelets are treated with a composition selected from the group consisting of thrombin, calcium chloride or its salts, thromboxane A2, adenosine triphosphate, arachidonate, and combinations thereof. 
     
     
         10 . The method of  claim 1 , further comprising:
 administering one or more compounds selected from the group consisting of growth factors, cytokines, integrins, cadherins, molecules or drugs that promote angiogenesis, molecules or drugs that promote vasculogenesis, molecules or drugs that promote aerteriogenesis, and combinations thereof.   
     
     
         11 . The method of  claim 10 , wherein the one or more compounds is selected from a group consisting of VEGF-A, PIGF, VEGF-B, VEGF-C, VEGF-D, TGF-β, Ang-1, Ang-2, IGF, HGF, FGF, Tie2, PDGF, CCL2, Alpha-V Beta-5, Alpha-5 Beta-1, VE-cadherin, PECAM-1, plasminogen activator, nitrogen oxide synthase, and combinations thereof. 
     
     
         12 . The method of  claim 1 , further comprising:
 administering one or more growth factors before, during, or after implanting the selected, viable, undifferentiated mesenchymal stem cells in the avascular zone.   
     
     
         13 . The method of  claim 12 , wherein the one or more growth factors is selected from the group consisting of TGF-β, FGF, PDGF, IGF, and combinations thereof 
     
     
         14 . The method of  claim 1 , further comprising:
 administering one or more compounds selected from the group consisting of growth factors, cytokines, integrins, cadherins, molecules or drugs that promote angiogenesis, molecules or drugs that promote vasculogenesis, molecules or drugs that promote aerteriogenesis, and combinations thereof.   
     
     
         15 . The method of  claim 1 , wherein the culturing is for multiple passages. 
     
     
         16 . The method of  claim 1 , wherein the culturing is for about two to about five passages. 
     
     
         17 . The method of  claim 1 , wherein the harvested nucleated cells are cultured at 6000 to 12,000 cells/cm 2 . 
     
     
         18 . The method of  claim 1 , wherein the harvested nucleated cells are cultured at less than or equal to 12,000 cells/cm 2 . 
     
     
         19 . A method for treating a degenerative intervertebral disc in a patient in need thereof, the method comprising:
 culturing harvested nucleated cells in a culture medium under a selective pressure of about 1% to about 10% oxygen and a pH of about 6.6 to about 7.0;   selecting viable, undifferentiated mesenchymal stem cells capable of growth in the culture medium under the selective pressure of about 1% to about 10% oxygen and a pH of about 6.6 to about 7.0; and   providing the selected, viable, undifferentiated mesenchymal stem cells for implantation in an intervertebral disc.   
     
     
         20 . A method for treating a degenerative intervertebral disc in a patient in need thereof, the method comprising:
 culturing harvested nucleated cells in a culture medium under a selective pressure of about 1% to about 10% oxygen and a pH of about 6.6 to about 7.0;   selecting viable, undifferentiated mesenchymal stem cells capable of growth in the culture medium under the selective pressure of about 1% to about 10% oxygen and a pH of about 6.6 to about 7.0; and   implanting the selected, viable, undifferentiated mesenchymal stem cells in an intervertebral disc.