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 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;   implanting the viable mesenchymal stem cells in a posterior disc annulus of the degenerative intervertebral disc; and   implanting autologous platelets or platelet lysate in the posterior disc annulus, either separately or in combination with the viable mesenchymal stem cells.   
     
     
         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 autologous platelets from the patient in need thereof. 
     
     
         6 . The method of  claim 1 , wherein the autologous platelets are implanted prior to, during or after implanting the viable mesenchymal stem cells. 
     
     
         7 . The method of  claim 5  wherein the autologous platelets are treated with thrombin and calcium chloride 1-7 days prior to implanting in the posterior disc annulus. 
     
     
         8 . The method of  claim 7  wherein the amount of thrombin is 28.56 U/ml and the amount of calcium chloride is 2.86 mg/ml. 
     
     
         9 . The method of  claim 5  wherein the autologous platelets are treated with thrombin, calcium chloride or its salts, thromboxane A2, adenosine triphosphate and arachidonate. 
     
     
         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, and molecules or drugs that promote arteriogenesis. 
     
     
         11 . The method of  claim 10  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. 
     
     
         12 . 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. 
     
     
         13 . The method of  claim 12  wherein the one or more growth factor is selected from the group consisting of: TGF-β, FGF, PDGF, and IGF. 
     
     
         14 . 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. 
     
     
         15 . A method for treating a degenerative intervertebral disc in a patient in need thereof comprising:
 culturing harvested nucleated cells in a culture medium 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;   providing the viable mesenchymal stem cells for implantation in a posterior disc annulus of the degenerative intervertebral disc; and   providing autologous platelets or platelet lysate for implantation in the posterior disc annulus.   
     
     
         16 . The method of  claim 15 , wherein the autologous platelets or platelet lysate is implanted either separately or in combination with the viable mesenchymal stem cells. 
     
     
         17 . The method of  claim 15  wherein the selective pressure further includes culturing the harvested nucleated cells in from about 2 to about 10% carbon dioxide. 
     
     
         18 . The method of  claim 15  wherein the autologous platelets are treated with thrombin and calcium chloride 1-7 days prior to implanting in the posterior disc annulus. 
     
     
         19 . The method of  claim 15  wherein the autologous platelets are treated with thrombin, calcium chloride or its salts, thromboxane A2, adenosine triphosphate and arachidonate. 
     
     
         20 . A method for treating an avascular zone in a patient in need thereof comprising:
 culturing harvested nucleated cells in a culture medium 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;   implanting the viable mesenchymal stem cells in the avascular zone; and   implanting autologous platelets or platelet lysate in the posterior disc annulus, either separately or in combination with the viable mesenchymal stem cells.