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目的:研究3种牙科金属材料(金合金、纯钛、镍铬合金)浸提液对小鼠成纤维细胞L929的细胞毒性及凋亡相关基因与蛋白表达的影响,探讨3种牙科金属材料引起细胞凋亡的可能通路。方法:以金合金(A组)、纯钛(B组)、镍铬合金(C组)的浸提液(每组8个样本)培养小鼠成纤维细胞L929,以含10%胎牛血清的RPMI1640培养基作为阴性对照(D组),以铜合金作为阳性对照(E组)。采用MTT法在细胞水平评价3种牙科金属材料的细胞毒性,采用RT-PCR在分子水平上检测3种牙科金属材料浸提液对L929细胞凋亡相关基因caspase-3、-8、-9 mRNA及蛋白表达的影响。采用SPSS11.5软件包对数据进行统计学分析。结果:各实验组细胞毒性均为0级。镍铬合金组caspase-3mRNA与caspase-9mRNA的表达与其他实验组、阴性对照及阳性对照组间差异显著,镍铬合金组caspase-3蛋白与caspase-9蛋白的表达与其他实验组及阴性对照组间差异显著,各组caspase-8mRNA及蛋白的表达无显著差异。结论:金合金与纯钛浸提液对小鼠L929细胞凋亡相关基因及蛋白的表达无显著诱导作用;镍铬合金诱导小鼠L929细胞凋亡相关基因caspase-3mRNA、caspase-9mRNA及蛋白表达增加,可能通过线粒体通路诱导细胞凋亡。
OBJECTIVE: To study the cytotoxicity and apoptosis related gene and protein expression of three kinds of dental metal materials (gold alloy, pure titanium and nickel-chromium alloy) on mouse fibroblast L929 cells and explore the effects of three kinds of dental metal materials Possible pathway of apoptosis. Methods: Mouse fibroblasts L929 were cultured in the leaching solution of gold alloy (group A), pure titanium (group B) and nickel-chromium alloy (group C) (8 samples per group), with 10% fetal bovine serum Of RPMI1640 medium as a negative control (group D), with copper alloy as a positive control (group E). MTT assay was used to evaluate the cytotoxicity of three dental materials at cellular level. The effects of three dental material extracts on the expression of caspase-3, -8, -9 mRNA in L929 cells were detected by RT-PCR And protein expression. SPSS11.5 software package for statistical analysis of the data. Results: The cytotoxicity of each experimental group was grade 0. Ni-Cr alloy caspase-3mRNA and caspase-9mRNA expression and other experimental groups, negative control and positive control group significant difference, Ni-Cr alloy caspase-3 protein and caspase-9 protein expression compared with other experimental group and negative control There was significant difference between the two groups. There was no significant difference in the expression of caspase-8mRNA and protein in each group. CONCLUSION: There is no significant induction of the expression of apoptosis-related genes and proteins in L929 cells induced by gold alloy and pure titanium; the expression of caspase-3 mRNA and caspase-9 mRNA and protein in L929 cells induced by Ni-Cr alloy Increased, may induce apoptosis through the mitochondrial pathway.