10-MDP钙盐对小鼠成纤维细胞L929增殖、凋亡及MMPs表达的影响

Effects of 10-MDP calcium salt on proliferation,apoptosis and MMPs expression of L929 fibroblasts

目的通过10-MDP钙盐对小鼠成纤维细胞L929的研究调查了不同浓度不同结构的10-MDP钙盐对牙周组织的影响。方法通过X射线衍射(X-Ray diffraction,XRD)对合成的三种不同比例10-MDP钙盐进行鉴定。使用CCK-8法检测10-MDP钙盐对小鼠成纤维细胞L929的细胞增殖毒性;流式细胞术检测细胞凋亡;蛋白免疫印迹实验检测细胞基质金属蛋白酶(matrix metalloproteinase,MMP)2、MMP9表达。结果XRD的结果证实三种合成方式均有不同结构10-MDP钙盐的产生;细胞增殖毒性CCK-8实验中仅有1.0 mg/mL MDP-1组在72 h后引起细胞增殖活性降低;各组培养72 h后均不诱导细胞凋亡;0.2 mg/mL和0.1 mg/mL MDP-2、MDP-1组均抑制L929细胞的MMP2和MMP9蛋白分泌。结论10-MDP钙盐可能对牙周组织中成纤维细胞没有细胞毒性作用,并且可以通过抑制MMP2、MMP9蛋白的分泌减少牙周组织破坏。

Objective To investigate the effects of different concentrations of 10-MDP calcium salts on biological behaviors of L929 fibroblasts in the periodontium.Methods The X-Ray diffraction(XRD)was used to examine 10-MDP calcium salts obtained by the three synthesized formulas.The cell proliferation and cytotoxicity of L929 was measured by CCK-8 method;the flow cytometry was used to detect cell apoptosis,and the protein expression of matrix metalloproteinase(MMP)-2 and MMP-9 was measured by Western blot.Results The XRD results confirmed that the 10-MDP calcium salts obtained by the three synthesized formulas were in different structures.In the CCK-8 assay,only the 1.0 mg/mL MDP-1 group caused a decrease in cell proliferation activity after 72 h.No cell apoptosis was induced in each group after 72 hours of culture.The secretion of MMP2 and MMP9 proteins in L929 cells were inhibited in 0.2 mg/mL and 0.1 mg/mL MDP-2 and MDP-1 groups.Conclusion 10-MDP calcium salts may not have a cytotoxic effect on the periodontium;they can exert an inhibitory effect on the destruction of the periodontium by inhibiting the secretion of MMP2 and MMP9.