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.

1 . A method for treating a degenerative intervertebral disc in a patient in need thereof comprising:
 harvesting stem cells from the patient in need thereof;   culturing the harvested stem cells under selective pressure that corresponds to the environment of a damaged intervertebral disc; and   implanting the selected stem cells in a posterior disc annulus of the degenerative intervertebral disc.   
     
     
         2 . The method of  claim 1  wherein the selective pressure include culturing the harvested stem cells in from about 3 to about 10% oxygen. 
     
     
         3 . The method of  claim 2  wherein the selective pressure includes culturing the harvested stem cells in from about 3 to about 7% oxygen. 
     
     
         4 . The method of  claim 1  wherein the selective pressure include culturing the harvested stem cells in from about 2 to about 10% carbon dioxide. 
     
     
         5 . The method of  claim 2  further comprising the selective pressure including culturing the harvested stem cells in from about 2 to about 10% carbon dioxide. 
     
     
         6 . The method of  claim 1  further comprising:
 harvesting NP cells from the patient in need thereof; and 
 co-culturing the NP cells with the harvested stem cells under selective pressure. 
 
     
     
         7 . The method of  claim 1  wherein the stem cells are mesenchymal stem cells. 
     
     
         8 . 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 selected stem cells; 
 wherein the implanting of both platelets and stem cells is in the posterior disc annulus. 
 
     
     
         9 . The method of  claim 8  wherein the platelets are treated with thrombin and calcium chloride 1-7 days prior to implanting in posterior disc annulus. 
     
     
         10 . The method of  claim 9  wherein the amount of thrombin is 28.56 U/ml and the amount of calcium chloride is 2.86 mg/ml. 
     
     
         11 . The method of  claim 8  wherein the platelets are treated with thrombin, calcium chloride or its salts, thromboxane A2, adenosine triphosphate and arichidonate. 
     
     
         12 . 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 aerteriogenesis. 
     
     
         13 . The method of  claim 12  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. 
     
     
         14 . The method of  claim 1  further comprising administering one or more growth factors before, during or after implanting selected stem cells in the posterior disc annulus. 
     
     
         15 . The method of  claim 14  wherein the one or more growth factor is selected from the group consisting of: TGF-β, FGF, PDGF, and IGF. 
     
     
         16 . The method of  claim 1  wherein the culturing of harvested stem cells is performed in a basal culture media made from a D-MEM base. 
     
     
         17 . A pharmaceutical composition comprising stem cells, platelets, one or more growth factors, and a pharmaceutical carrier or diluent. 
     
     
         18 . The composition of  claim 17  wherein the stem cells are mesenchymal stem cells. 
     
     
         19 . The composition of  claim 17  wherein the stem cells have been selected for their capacity for viability and expansion under hypoxic conditions. 
     
     
         20 . The compositions of  claim 17  wherein the stem cells have been selected for their capacity for viability and expansion under culture conditions comparable to a damaged intervertebral disc. 
     
     
         21 . A method for treating an avascular zone in a patient in need thereof comprising:
 harvesting stem cells from the patient in need thereof;   culturing the harvested stem cells under selective pressure that corresponds to the environment of a damaged or aged avascular zone; and   implanting the selected stem cells in the avascular zone.   
     
     
         22 . The method of  claim 21  wherein the avascular zone is in a patient's shoulder. 
     
     
         23 . The method of  claim 21  wherein the avascular zone is in a patient's hip.