Method for in vivo, ex vivo and in vitro repair and regeneration of cartilage and collagen and bone remodeling

Abstract

A method for in vivo, ex vivo and in vitro regeneration of cartilage and collagen. In vivo, ex vivo and in vitro regeneration and de novo formation of articular cartilage and collagen by intermittently applied hydrostatic pressure. The application of external interval loading consisting of repeated periods of applied hydrostatic pressure followed and interrupted by periods of recovery. The application of the intermittent hydrostatic pressure at physiological levels 5-10 MPA for an interval of 4 hours followed by a recovery period up to about 20 hours, said pressure applied to the cartilage cells in vitro, explants of cartilage ex vivo and in vivo to cartilage that remains intact within te joint space of diarthrotic joints. The interval loading results in the selective inhibition of matrix degrading enzymes, pro-inflammatory cytokines and chemokines that attract inflammatory cells into the joint cavity and in selective decrease of gene expression of growth factors that are inhibitory to type II collagen expression.

1 - 73 . (canceled) 
   
   
       74 . A method of increasing formation of extracellular matrix by increasing production of aggrecan and Type II collagen, said method comprising a step of treating adult human chondrocytes in vitro or ex vivo with an intermittently applied hydrostatic pressure of between about 0.5 and about 30 MPa, at a frequency of about 0.1 to about 10 Hz for a period of about 1 to about 8 hours, followed with a constant atmospheric pressure applied for a period of about 16 to 23 hours, wherein said period of hydrostatic pressure followed by the recovery period is repeated for about 4 to about 100 days. 
   
   
       75 . The method of  claim 74  wherein said human chondrocytes are normal healthy chondrocytes. 
   
   
       76 . The method of  claim 75  wherein said human chondrocytes are osteoarthritic chondrocytes. 
   
   
       77 . The method of  claim 76  wherein said osteoarthritic chondrocytes are metabolically inactive. 
   
   
       78 . The method of  claim 77  wherein said osteoarthritic chondrocytes are isolated from injured, diseased or aged articular cartilage. 
   
   
       79 . The method of  claim 78  suitable for treatment of an articular cartilage injury and repair or for cartilage regeneration. 
   
   
       80 . The method of  claims 74  wherein said method is used for increased production of extracellular matrix within a cartilage graft. 
   
   
       81 . The method of  claim 80  wherein said graft subjected to treatment of  claim 74  is used as an implant for treatment of cartilage injury or repair of cartilage. 
   
   
       82 . Activated osteoarthritic metabolically inactive chondrocytes isolated from a diseased, aged or injured articular cartilage pressure treated in vitro or ex vivo with an intermittently applied hydrostatic pressure of between about 0.5 and about 30 MPa, at a frequency of about 0.1 to about 10 Hz for a period of about 1 to about 8 hours, followed with a constant atmospheric pressure applied for a period of about 16 to 23 hours, wherein said period of hydrostatic pressure followed by the recovery period is repeated for about 4 to about 100 days, said chondrocytes suitable for treatment of osteoarthritic defects or cartilage injuries. 
   
   
       83 . The activated chondrocytes of  claim 82  wherein said treatment increases levels of aggrecan from initial levels between about 0.1 and 1%, wet weight, and type II collagen between about 1 and 10%, wet weight. 
   
   
       84 . The activated chondrocytes of  claim 83  wherein said levels of aggrecan and Type II collagen are increased to levels of about 4 and about 7% and levels of type II collagen are increased to about 10 and 20%. 
   
   
       85 . The activated chondrocytes of  claim 84  submitted to said treatment until said levels of aggrecan or Type II collagen reach levels of active chondrocytes.