Patent ID: 6464729
Filing Date: 2002-10-15
Classification: A61K,A61L,A61P,C12N,Y10S

Abstract:
A method for producing an isolated mineralized biological material comprising a continuous layer of mineralization comprising chondrocytes having alkaline phosphatase activity and surrounded by matrix containing type I, type II and type X collagens, sulphated proteoglycans having a large hydrodynamic size, matrix vesicles and calcium hydroxyapatite crystal deposits, comprising:(a) isolating chondrocytes from the lower 15% of animal articular cartilage tissue having an alkaline phosphatase activity of at least 2 &mgr;M PNP/hr/&mgr;g DNA; (b) forming a layer of the chondrocytes on a substrate wherein the substrate is bone, an engineered biomaterial or a porous tissue culture insert; (c) culturing the chondrocytes in growth media under suitable conditions so that the chondrocytes accumulate matrix and form cartilaginous tissue which contains components associated with cartilage mineralization, and (d) culturing the cartilaginous tissue in the presence of a mineralizing agent selected from the group consisting of &bgr;-glycerophosphate, ATP, and phosphoethanolamine, to form the mineralized biological material.