Effects Of B-FGF And BMP-2 On Proliferation And Differentiation Of Rat Articular Chondrocytes Cultured In Vitro

ObjectiveTo study the effects of basic fibroblast growth factor (b-FGF) and Bone Morphogenetic Protein2 (BMP-2) on the proliferation and differentiation of articular chondrocytes cultured in monolayer,and search for the optimal concentration of b-FGF and BMP-2 applied vitro in order to improve the culture condition of tissue-engineered cartilage regeneration.MethodsArticular chondroeytes of Wistar rats were isolated and cultured in vitro. Make the third generation of chondroeytes cell which at good function as the experimental distribution after the added different combinations and concentrations of growth factor. A group is set to blank control group, the experimental group according to b-FGF, and BMP-2 concentrations were 6 groups, the concentration of the two factors were 1ng/ml, 3ng/ml, 5ng/ml, 10ng/ml, 20ng/ml , 30ng/ml. We measured the OD value in 490nm of each groups convert to cell relative numbers by MTT method in 24H,3days,5days after growth factors were given. Studied the expressions of collagen typeâ…¡and semiquantitative analysis of cell climbing sheets which gathered after 72h by immunocytochemistry . Two factors were obtained on the role of chondrocyte proliferation and differentiation of the optimal concentration, and then alone or in combination with the optimal concentration of culture are two factors come to a synergistic effect. ResultsB-FGF (5-30ng/ml) could promote chondrocyte proliferation, especially chondroeytes cell measured the crest OD value at dose of 20ng/ml. BMP-2 concentrations on the OD value of chondrocytes were not significantly increased relative to other. b-FGF on chondrocyte synthesis of collagen typeâ…¡had no effect, and BMP-2 can significantly promote collagen synthesis of cartilage cells to 20ng/ml for the best concentration. Optimal concentration of the two factors combined effects of cartilage cells, the OD value and the average gray value of the contrast in the other groups were statistically significant.ConclusionsB-FGF could obviously promote prophase chondrocyte proliferation contrast to anaphase of chondrocyte. BMP-2 could also promote chondrocytes proliferation, secretion of collagen typeâ…¡and maintenance of cell phenotype, the both factors have synergetic reaction.