聚甲基丙烯酸甲酯/矿化胶原/Mg-Ca合金复合材料与小鼠前成骨细胞的成骨分化  

Effect of polymethyl methacrylate/mineralized collagen/Mg-Ca composite material on osteogenic differentiation of mouse preosteoblasts

作　　者：张红梅 孙溪饶 王程越 Zhang Hongmei;Sun Xirao;Wang Chengyue(Second Affiliated Hospital of Jinzhou Medical University,Jinzhou 121000,Liaoning Province,China;Affiliated Hospital of Chifeng College,Chifeng 024000,Inner Mongolia Autonomous Region,China)

机构地区：[1]锦州医科大学附属第二医院,辽宁省锦州市121000 [2]赤峰学院附属医院,内蒙古自治区赤峰市024000

出　　处：《中国组织工程研究》2022年第16期2498-2503,共6页Chinese Journal of Tissue Engineering Research

基　　金：辽宁省教育厅重点攻关项目(JYTZD2020004),项目负责人:王程越;辽宁省自然基金资助计划(2019-MS-141),项目负责人:孙溪饶。

摘　　要：背景:课题组前期通过冷冻干燥方法制备的矿化胶原/Mg-Ca合金复合体结构松散,使用聚甲基丙烯酸甲酯骨水泥作为外层加强层对其进行改良可增强复合材料的结构稳定性。目的:探讨聚甲基丙烯酸甲酯/矿化胶原/Mg-Ca合金复合材料对MC3T3-E1细胞成骨分化的影响。方法:分别制备聚甲基丙烯酸甲酯/矿化胶原/Mg-Ca合金、聚甲基丙烯酸甲酯/矿化胶原复合材料。将MC3T3-E1细胞分别接种于聚甲基丙烯酸甲酯/矿化胶原/Mg-Ca合金(A组)、聚甲基丙烯酸甲酯/矿化胶原复合材料(B组)及Mg-Ca合金材料(C组)上,以单独培养的细胞为空白对照(D组),扫描电镜下观察材料表面的细胞黏附形态,通过CCK-8、碱性磷酸酶活性、酶联免疫吸附实验、RT-PCR与Western blot检测分析3种材料对MC3T3-E1细胞增殖及成骨分化的影响。结果与结论:(1)扫描电镜下可见,MC3T3-E1细胞在A组复合材料表面黏附数量最多,生长状态良好,细胞拉伸明显,伪足伸出较多,细胞相互连接成网状结构;(2)CCK-8实验显示,与D组相比,3种材料均可促进MC3T3-E1细胞的增殖(P<0.05),其中以A组复合材料促增殖效果最显著(P<0.05);(3)与D组相比,3种材料均可提升MC3T3-E1细胞中的碱性磷酸酶活性(P<0.05),其中以A组提升最显著(P<0.05);(4)酶联免疫吸附实验显示,3种材料均可促进MC3T3-E1细胞分泌骨钙素与I型胶原,其中以A组促进作用最显著(P<0.05);(5)RT-PCR检测显示,3种材料均可提高MC3T3-E1细胞内RUNX2、OSX、碱性磷酸酶、骨钙素的m RNA表达,其中以A组提高最显著(P<0.05);(6)Western blot检测显示,3种材料均可提高MC3T3-E1细胞内RUNX2、OSX、碱性磷酸酶、骨钙素的蛋白表达,其中以A组提高最显著(P<0.05);(7)结果表明,聚甲基丙烯酸甲酯/矿化胶原/Mg-Ca合金复合材料可促进MC3T3-E1细胞成骨分化相关蛋白与基因的表达,对其成骨分化起积极作用。BACKGROUND: The structure of mineralized collagen/Mg-Ca composite prepared by freeze-drying method is loose, and the structural stability of the composite can be enhanced by using polymethyl methacrylate bone cement as the outer strengthening layer.OBJECTIVE: To investigate the effects of polymethyl methacrylate/mineralized collagen/Mg-Ca composite on osteogenic differentiation of MC3 T3-E1 cells. METHODS: The polymethyl methacrylate/mineralized collagen/Mg-Ca and polymethyl methacrylate/mineralized collagen composites were prepared respectively. The MC3 T3-E1 cells were directly inoculated on the surface of polymethyl methacrylate/mineralized collagen/Mg-Ca composite(group A), polymethyl methacrylate/mineralized collagen composite(group B) and Mg-Ca alloy(group C). Cells cultured alone were blank controls(group D). The cell adhesion morphology on the material surface was observed under the scanning electron microscope. CCK-8, alkaline phosphatase activity, enzyme linked immunosorbent assay, RT-PCR, and western blot assay were used to analyze the effects of three materials on MC3 T3-E1 cell proliferation and osteogenic differentiation. RESULTS AND CONCLUSION:(1) Scanning electron microscope showed that the largest adhesion number of MC3 T3-E1 cells on the surface of polymethyl methacrylate/mineralized collagen/Mg-Ca composite, grew well, stretched obviously, pseudopodia extended more, and connected with each other to form a network structure.(2) CCK-8 assay showed that compared with the group D, the three materials could promote the proliferation of MC3 T3-E1 cells(P < 0.05), and the composite had the most significant effect in group A(P < 0.05).(3) Compared with the group D, the three materials could promote the alkaline phosphatase activity in MC3 T3-E1 cells(P < 0.05), and the group A was the most significant(P < 0.05).(4) Enzyme linked immunosorbent assay showed that the three materials could promote the secretion of osteocalcin and type I collagen in MC3 T3-E1 cells, and the promotion effect of group A was t

关 键 词：聚甲基丙烯酸甲酯骨水泥 矿化胶原 成骨分化 镁钙合金 小鼠前成骨细胞 转录因子 骨钙素 碱性磷酸酶 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学]