【摘要】 目的 探讨锶离子(Sr²⁺)对聚甲基丙烯酸甲酯（polymethyl methacrylate, PMMA）颗粒环境中成骨细胞（osteoblast, OB）增殖、分化的作用及机制。方法 选取小鼠颅顶前骨细胞亚克隆14（MC3T3-E1 Subclone 14,MC3T3-E1）为研究对象，分为空白组、PMMA组、Sr²⁺组。细胞增殖-毒性检测试剂盒（cell proliferation-toxicity Assay Kit 8,CCK-8）检测1 g/L PMMA颗粒和（0.5 mM、1 mM、3 mM、10 mM）Sr²⁺处理MC3T3-E1细胞24 h、48 h、72 h后的增殖活性；酶联免疫吸附试验（enzyme-linked immunosorbent assay, ELISA）检测细胞碱性磷酸酶（alkaline phosphatase, ALP）活性；茜素红染色检测细胞基质矿化程度；实时定量聚合酶链反应（reverse transcription-polymerase chain reaction, RT-PCR）检测RUNT相关转录因子2（RUNT-related transcription factor 2,Runx-2）基因表达水平；蛋白免疫印迹（Western blot, WB）检测整合素α2（integrin α2,ITGα2）、黏着斑激酶（plaque adhesion kinase, FAK）、磷酸化FAK（p-FAK）蛋白相对表达量。结果 与PMMA组比较，3 mM Sr²⁺组能够促进MC3T3-E1细胞增殖，上调细胞ALP活性、钙结节数量以及Runx-2 mRNA水平，增加MC3T3-E1细胞中ITGα2蛋白表达量和p-FAK/FAK蛋白表达量比值，差异均有统计学意义（P<0.05）。结论 Sr²⁺通过上调ITGα2/FAK信号通路促进PMMA颗粒环境中OB活化，Sr²⁺有望成为解决关节假体无菌性松动的重要手段。更多还原

【Abstract】 Objective To investigate the effects and mechanism of strontium ions(Sr²⁺) on the proliferation and differentiation of osteoblasts（OB） in the granular environment of polymethyl methacrylate（PMMA）.Methods Mouse cranial anterior parietal osteocytes（MC3T3-E1） were selected as the research object, and were divided into the blank group, the PMMA group, and the Sr²⁺ group. The cell proliferation-toxicity assay kit（CCK-8） was used to detect the proliferation activity of 1 g/L PMMA particles and MC3T3-E1 cells treated with（0.5 mM, 1 mM, 3 mM, 10 mM） Sr²⁺ for 24 h, 48 h, and 72 h; the activity of alkaline phosphatase in cells was detected by enzyme-linked immunosorbent assay（ELISA）; alizarin red staining was used to detect the degree of cell matrix mineralization; the expression level of RUNT-related transcription factor 2（Runx-2） gene was detected by reverse transcription-polymerase chain reaction（RT-PCR）; the relative expression levels of integrin α2（ITGα2）, plaque adhesion kinase（FAK）, and phosphorylated FAK（p-FAK） proteins were detected by Western blot（WB）.Results Compared with the PMMA group, the 3 mM Sr²⁺ group could promote the proliferation of MC3T3-E1 cells, up-regulate the ALP activity, the number of calcium nodules and the Runx-2 mRNA level of the cells, increase the expression level of ITGα2 protein and the ratio of p-FAK/FAK protein in MC3T3-E1 cells, and the differences were statistically significant（P<0.05）.Conclusion Sr²⁺ promotes OB activation in the PMMA particle environment by upregulating the ITGα2/FAK signaling pathway, and Sr²⁺ is expected to become an important means to solve aseptic loosening of joint prostheses.更多还原