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 a degenerative intervertebral disc in a patient in need thereof comprising:
 culturing harvested nucleated cells in a culture medium supplemented with 10-20% platelet cell lysate under selective pressure of about 1% to about 10% oxygen for 1-28 days;   obtaining viable mesenchymal stem cells capable of growth in the culture medium under selective pressure of about 1% to about 10% oxygen; and   implanting the viable mesenchymal stem cells in a posterior disc annulus of the degenerative intervertebral disc.   
     
     
         2 . The method of  claim 1  wherein the selective pressure includes culturing the harvested nucleated cells in from about 3 to about 7% oxygen. 
     
     
         3 . The method of  claim 1  wherein the selective pressure further includes culturing the harvested nucleated cells in from about 2 to about 10% carbon dioxide. 
     
     
         4 . The method of  claim 2  wherein the selective pressure further includes culturing the harvested nucleated cells in from about 2 to about 10% carbon dioxide. 
     
     
         5 . 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 viable mesenchymal stem cells; 
 wherein the implanting of both platelets and viable mesenchymal stem cells is in the posterior disc annulus. 
 
     
     
         6 . The method of  claim 5  wherein the platelets are treated with thrombin and calcium chloride 1-7 days prior to implanting in the posterior disc annulus. 
     
     
         7 . The method of  claim 6  wherein the amount of thrombin is 28.56 U/ml and the amount of calcium chloride is 2.86 mg/ml. 
     
     
         8 . The method of  claim 5  wherein the platelets are treated with thrombin, calcium chloride or its salts, thromboxane A2, adenosine triphosphate and arachidonate. 
     
     
         9 . 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, and molecules or drugs that promote arteriogenesis. 
     
     
         10 . The method of  claim 9  wherein the compound is 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, or nitrogen oxide synthase. 
     
     
         11 . The method of  claim 1  further comprising administering one or more growth factors before, during or after implanting the viable mesenchymal stem cells in the posterior disc annulus. 
     
     
         12 . The method of  claim 11  wherein the one or more growth factor is selected from the group consisting of: TGF-β, FGF, PDGF, and IGF. 
     
     
         13 . The method of  claim 1  wherein the culturing of harvested nucleated cells is performed in a basal culture media made from a D-MEM base. 
     
     
         14 . The method of  claim 3  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, and molecules or drugs that promote arteriogenesis. 
     
     
         15 . The method of  claim 3  further comprising administering one or more growth factors before, during or after implanting the viable selected mesenchymal stem cells in the posterior disc annulus. 
     
     
         16 . The method of  claim 5  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, and molecules or drugs that promote arteriogenesis. 
     
     
         17 . The method of  claim 5  further comprising administering one or more growth factors before, during or after implanting the viable selected mesenchymal stem cells in the posterior disc annulus. 
     
     
         18 . A method for treating a degenerative intervertebral disc in a patient in need thereof comprising:
 culturing harvested nucleated cells in a culture medium supplemented with 10-20% platelet cell lysate under selective pressure of about 3 to about 10% oxygen for two to five passages;   obtaining viable mesenchymal stem cells capable of growth in the culture medium under selective pressure of about 3% to about 10% oxygen; and   providing the viable mesenchymal stem cells for implantation in a posterior disc annulus of the degenerative intervertebral disc.   
     
     
         19 . The method of  claim 18  wherein the selective pressure further includes culturing the harvested nucleated cells in from about 2 to about 10% carbon dioxide. 
     
     
         20 . A method for treating an avascular zone in a patient in need thereof comprising:
 culturing harvested nucleated cells in a culture medium supplemented with 10-20% platelet cell lysate under selective pressure of about 3% to about 10% oxygen for 1-28 days;   obtaining viable mesenchymal stem cells capable of growth in the culture medium under selective pressure of about 3% to about 10% oxygen; and   implanting the viable mesenchymal stem cells in the avascular zone.