Effects of fibrin gel combined with basic fibroblast growth factor on alkaline phosphatase activity in fetal rat limb cells / 中国组织工程研究

ABSTRACT

BACKGROUND: Fibrin is a natural biodegradable polymer material with good histocompatibility. Basic fibroblast growth factor (bFGF) is an important polypeptide growth factor with mitogenic effect. Fibrin gel (FG) combined with bFGF is conducive to cell migration and promotes proliferation and differentiation. OBJECTIVE: To investigate the effects of fibrin gel combined with bFGF on alkaline phosphatase activity (ALP) in fetal rat limb cells. METHODS: Four groups were included FG group fetal rat limb cells were seeded in FG (three-dimensional, 3D) with normal medium; FG + bFGF group cells were seeded in FG containing 10 μg/L bFGF for 24-hour 3D culture; bFGF group cells were seeded in 10 μg/L bFGF medium; normal control group cells were seeded in normal culture medium. Under aseptic condition, rat fetal limb cells were isolated. Passage 3 cells were used for each group above mentioned. ALP activity, mRNA expression and cell morphology were analyzed at 3, 5, 7, 14, 21, and 28 days after culture. RESULTS AND CONCLUSION: The cells in the FG had more processes and formed an interconnected network after 7 days of incubation, while the cells in no gels remained cuboidal or "spindle-shaped". At 7, 14, and 21 days, ALP activity was greater in the FG+ bFGF group than in the other groups (P < 0.05). At each time point, mRNA expression of ALP was greater in the FG+ bFGF group than in the control group (P < 0.01). These findings indicate that FG combined with bFGF is conducive to cell morphogenesis and markedly enhances ALP activity.