Effects of bFGF and Collagen on the Construction of Tissue Engineering Cartilage In Vitro in the 3-D Culture System

Abstract:

Objective: To investigate effects of bFGF and collagen on tissue-engineered cartilage in pellet culture. Methods: Rabbit rib growth-plate chondrocytes were incubated in pellet culture. bFGF and Collagen was applied alone and in combination based on the results of previous search . The histological structure of neocartilage was observed by species stained with H＆E and Immunohistochemical analysis. The DNA content of neocartilage was measured by hoechst33258 dye. The production of matrix was estimated from the determination of hydroxyproline content and Alcian Blue method. Results: Chondrocytes formed cartilage-like tissue with distinct differentiated zones and regions. The DNA content of neocartilage was enhanced by bFGF and collagen. The production of collagen and PG was promoted by collagen. bFGF and collagen could suppress the expression of collagenⅠwhile they could decelerate the decrease of collagenⅡ. The combination of bFGF and collagen had synergistic effects. Conclusion: Pellet culture could maintain chondrocytes phenotype and enhance matrix production; bFGF could stimulate the proliferation of chondrocytes, collagen could stimulate the proliferation and the matrix production of chondrocytes, it also had a positive effect on the maintenance of cell phenotype, the combination of bFGF and collagen had synergistic effects on the maintenance of chondrocytes phenotype and the regeneration of cartilage.