氟对体外培养大鼠成釉细胞活性和细胞凋亡的影响

Effect of fluoride on the viability and apoptosis of ameloblasts in vitro

目的:观察氟对体外培养的大鼠成釉细胞活性的影响,为探讨氟斑牙的形成机制提供依据。方法 :取对数生长期的大鼠成釉细胞,在培养液中加入浓度为0、0.4、0.8、1.6、3.2、6.4 mmol/L的NaF溶液,培养24、48、72 h后,采用CCK-8检测各组细胞的活性。荧光显微镜下观察成釉细胞核形态的变化,流式细胞术分析氟对细胞凋亡的影响。采用SPSS13.0软件包对数据进行统计学分析。结果:1NaF浓度为0.4、0.8 mmol/L时,对成釉细胞有促增殖作用;NaF浓度为1.6、3.2、6.4 mmol/L时,对成釉细胞的活性有抑制作用,随着NaF浓度的增加,对细胞的抑制作用也逐渐增强,并且这种双向调节作用呈时间依赖性。2NaF浓度为0.4、0.8 mmol/L时,鲜见核破碎;NaF浓度为1.6、3.2 mmol/L时,存在核破碎,并且随着浓度的提高,核破碎的数量随之增加,即1.6 mmol/L浓度的NaF可引起成釉细胞凋亡,随着浓度的增加,细胞凋亡的数量随之增加。结论:1氟对体外培养成釉细胞的增殖具有双向调节作用,即低浓度促进,高浓度抑制。2浓度超过1.6 mmol/L时,NaF诱导成釉细胞凋亡。

PURPOSE： To evaluate the effect of fluoride on viability of rat ameloblasts in vitro. METHODS： The ameloblasts of rat was exposed to different concentrations of NaF （0, 0.4, 0.8, 1.6, 3.2, 6.4 mmol/L） for 24, 48 and 72 hours. CCK-8 assays were performed to measure the cells proliferation; The morphology of apoptosis was observed by Hoechst 33258 staining and the rate of apoptosis was determined by flow cytometry. The data was analyzed using SPSS 13.0 software package. RESULTS： ①The proliferation of ameloblasts was increased when concentrations of NaF between 0.4 mmol/L and 0.8 mmol/L, whereas inhibited at 1.6 mmol/L NaF and above. The effects were in time-dependent manner.②Cells in the 1.6 mmol/L NaF groups showed unclear karyorrhexis and apoptotic cell morphology. The effects were in concentration- dependent manner. CONCLUSIONS： ①Fluoride has two-phase effects to ameloblasts： At low doses, it promoted cell proliferation while at high doses it had negative effects.②1.6 mmol/L NaF could induce apoptosis of ameloblasts. Supported by National Natural Science Foundation of China （81072245） and Natural Science Foundation of Liaoning Province （20102278）.