目的 观察氟对体外培养小鼠成骨细胞增殖的影响。方法 原代培养小鼠成骨细胞,以0(对照组)、5、10、20、40mg/L剂量染氟,光、电镜观察成骨细胞的形态学变化;采用生长曲线、噻唑蓝(MTT)法及流式细胞仪检测细胞周期,观察细胞数量变化和细胞周期时相分布。结果 第10天时,5、10、20mg/L染氟组从形态学上表现出细胞增殖。而40mg/L组中出现细胞凋亡。染氟24h时,20mg/L组与对照组相比细胞增殖明显,差异具有统计学意义(P〈0.05),随着时间延长,染氟组表现出更强的增殖活性,至72h时,各染氟组成骨细胞增殖均高于对照组(P〈0.05)。24h时各染氟组间细胞增殖水平未见明显差异(P〉0.05),48h以后40mg/L组明显低于其他各染氟组(P〈0.05)。而染氟72h时5mg/L组增殖强度也低于10、20mg/L组(P〈0.05)。与对照组相比,10、20mg/L组处于G0/G1期的细胞数减少,而S期细胞数增加(P〈0.05),同时两组的细胞增殖指数也高于对照组(P〈0.05),生长表现出增殖状态;其中10mg/L组与其他染氟组之间,差异有统计学意义(P〈0.05),表现为细胞增殖能力增强。结论 低剂量的氟在较短作用时间内可以促进体外培养小鼠成骨细胞增殖,随剂量增加和暴露时间延长其促进作用减弱。
Objective To observe the proliferation of in vitro cultured mice osteoblasts exposed to different does of fluoride. Methods After primary culture osteoblasts exposed to fluoride at concentrations of 0,5,10,20,40 mg/L for 10 days, the morphological changes was observed by HE staining and electronic microscope, the osteoblasts were countered, and the distribution of cell cycle phases was analyzed. Results Exposed to fluoride for 10 days, osteoblasts of 5, 10,20 mg/L groups morphologically proliferated, some of 40 mg/L group appeared apoptosis; after 24 hours, there was a significantly difference between 20 mg/L and control group(P 〈 0.05). Along with time goning, groups exposed to fluoride had stronger ability of proliferation that was stronger than that in control group(P 〈 0.05). The absorbance was not significantly different between all groups(P〉 0.05) exposed to fluoride for 24 hours, but that of 40 mg/L group were lower than others (P 〈 0.05) after 48 hours, and 5 mg/L group were lower than 10,20 mg/L groups after 72 hours. Compared to control group, osteoblast numbers of G0/G1 in phase 10,20 mg/L groups were decreased, but those of S phase were increased (P 〈 0.05), and same thing happened with proliferation index (PI); the ability of proliferation was stronger in, significantly different from others (P 〈 0.05). Conclusions Low dose of fluoride improves the proliferation of in vitro osteoblasts in a short time exposure, which will be weakened with dose of fluoride increasing and time of exposure prolonging.