釉基质蛋白对人牙周膜细胞生物学影响的体外研究

目的:了解釉基质蛋白对人牙周膜细胞的增殖、矿化、蛋白合成及超微结构的影响。方法:组织块法原代培养人牙周膜细胞,茜素红染色进行细胞表型的鉴定;采用细胞增殖试剂盒及流式细胞仪检测釉基质蛋白对人牙周膜细胞增殖及细胞周期的影响;BCA蛋白定量试剂盒分析釉基质蛋白对牙周膜细胞蛋白合成的影响;并用透射电镜观察牙周膜细胞超微结构的改变;免疫组化染色观察釉基质蛋白作用下对牙周膜细胞骨涎蛋白和骨桥蛋白表达的影响。结果:釉基质蛋白可促进牙周膜细胞增殖,促进牙周膜细胞DNA的合成,使牙周膜细胞周期的G1期细胞所占百分比降低,而S期细胞所占百分比升高,并可提高牙周膜细胞蛋白总量,透射电镜观察可见经釉基质蛋白作用后的牙周膜细胞胞浆丰富,与蛋白合成相关细胞器粗面内质网及高尔基体发达,釉基质蛋白可促进牙周膜细胞矿化相关蛋白骨涎蛋白和骨桥蛋白的表达。结论:釉基质蛋白能促进牙周膜细胞增殖、蛋白合成及矿化相关蛋白骨涎蛋白和骨桥蛋白的表达,促进牙周组织的再生。

Effects of emdogain on human periodontal ligament cells in vitro

Objective: To investigate the effects of emdogain,enamel matrix derivative(EMD),on the proliferation,cell cycle,mineralization and ultrastructure of human periodontal ligament(PDL) cells in vitro.Methods: The influence of cell growth on PDL cells was evaluated by Cell Counting Kit-8(CCK-8) in the presence and absence of emdogain,after 1,3,and 5 d of culture.DNA synthesis and ultrastructure of PDL cells were observed by flow cytometry(FCM) and transmission electron microscopy(TEM) in the presence and absence of emdogain after 3 d of culture.The increasing of osteogenic capa-city was verified by the expression changes of osteogenic differentiation markers of bone sialoprotein(BSP) and osteopontin(OPN) in emdogain-treated PDL cells by immunohistochemicl staining.Results: Incubation of PDL cells with emdogain after 3 d significantly stimulated cell growth and DNA synthesis.Emdogain enhanced the osteogenic potential of PDL cells by high expression of osteogenic diffe-rentiation markers of BSP and OPN.Conclusion: The data indicate that Emdogain enhances cell prolife-ration and promotes differentiation of PDL cells,which contributes to periodontal tissue regeneration.