Influence of basic fibroblast growth factor on proliferation and differentiation of growth plate chondrocytes / 中国组织工程研究

ABSTRACT

BACKGROUND: The proliferative capacity of in vitro cultured chondrocytes is gradually reduced following several passages. Many studies have demonstrated that basic fibrublast growth factor (bFGF) can promote chondrocyte proliferation, and influence its phenotype and differentiation. OBJECTIVE: To observe the influence of bFGF on the proliferation and differentiation of growth plate chondrocytes and to screen the best dosage for in vitro culture of chondrocytes. DESIGN, TIME AND SETTING: For observation and comparison of cells, the experiment was performed at Tissue Engineering Laboratory, Department of Biochemistry, Jinan University Medical School between October 2004 and February 2005. MATERIALS Twelve New Zealand rabbits were selected for chondrocyte isolation; bFGF was purchased from PeproTech, UK. METHODS: Rabbit growth plate chondrocytes were isolated and cultured in low serum. Different doses of bFGF (0, 1, 2.5, 5, 10, 25, 50 and 100 μg/L) were added. MAIN OUTCOME MEASURES: Cell proliferative multiple was determined using modified MTT method; chondrocyte collagen content was determined using hydroxyproline method; alkaline phosphatase activities were detected by enzyme kinetics. RESULTS: bFGF (5-100 μg/L) could promote chondrocyte proliferation, especially at dose of 25 μg/L. bFGF at dose of larger than 25 μg/L would inhibit collagen production; bFGF at dose of larger than 1 μg/L would inhibit alkaline phosphatase activities. CONCLUSION: bFGF can stimulate growth plate chondrocyte proliferation, and inhibit growth plate chondrocyte differentiation at dose25 μg/L.