促矿化液对大鼠成骨细胞增殖分化的影响

Effects of mineralization liquid on rat′s osteoblast proliferation and differentiation

目的比较大鼠成骨细胞在矿化条件与非矿化条件下增殖和分化的过程,评价促矿化液对其生理功能的影响,明确促矿化液加入细胞培养环境的适宜时间。方法取SD大鼠头盖骨做成骨细胞原代培养,将增殖稳定后的第4代细胞分别在矿化条件和非矿化条件下培养,检测其形态、碱性磷酸酶活性、细胞周期等。结果大鼠成骨细胞增殖基本稳定后促矿化液组与无促矿化液组细胞分裂增殖指数相似,但前者碱性磷酸酶活性明显较高且持久。结论成骨细胞增殖基本稳定后加入促矿化液对细胞增殖无影响,但可明显促进细胞矿化功能,是加入促矿化液诱导细胞矿化功能的较好时机。

Objective To compare the rat osteoblast proliferation and differentiation in the mineralization condition and nonineralization condition, and to prove the feasible time of adding the mineralization liquid to the cell surroundings. Methods SD rat＇s calvarial osteoblast was primary cultured and passaged to the 4^th generation. After proliferation stabilized, the 4^th generation rat＇s osteoblast was cultured in the mineralization condition and nonmineralization condition. The cellular modality was observed by inverted microscopy analyzing system. Proliferation was described by MTT chromatometry and growth curve. Expression of alkaline phosphatase （ALP） was proved by alkaline phosphatase staining, mineralized nodus alizarin red staining and enzyme-linked immunosorbent assay （ELISA）. The cell apoptosis and proliferation were examined by flow cytomctry. Results The proliferation of osteo- blast went to the flat stage about in the 8^th day. After in the mineralization condition 3 weeks, a number of mineralized nodus formed. After the osteoblast proliferation almost stabilized, the mineralization group and the nonmineralization group had the similar proliferation index （PI）, but in the mineralization group, the expression of alkaline phosphatase of the osteoblast was more and kept longer than the last group. Conclusion After the osteoblast proliferation almost stabilized, adding the mineralization liquid would not influence the proliferation, but accelerate the osteoblast alkaline phosphatase expression and mineralized nodus amount. It proved that the feasible time to add the mineralization liquid was after the cell proliferation almost stabilized.