【摘要】 目的 评估低氧预处理的骨髓间充质干细胞(BMSCs)细胞对小鼠颅顶前骨细胞(MC3T3-E1细胞)生物学活性和成骨分化的影响。方法 将低氧诱导的BMSCs无血清培养基上清加入培养的MC3T3-E1细胞作为低氧组,常氧诱导的BMSCs无血清培养基上清加入培养的MC3T3-E1细胞作为常氧组,正常培养的细胞为对照组。在培养的第1、3、5和7天,CCK8检测细胞增殖活性,生化试剂盒检测碱性磷酸酶(ALP)含量,qRT-PCR检测Runt相关转录因子2(Runx2)、牛骨钙素(OCN)及骨桥蛋白(OPN)的mRNA表达水平,Western blot检测血管内皮生长因子(VEGF)及骨形态发生蛋白2型(BMP-2)的蛋白表达。结果 处理后第1、3、5和7天,与对照组相比,低氧组细胞增殖能力增强(P＜0.05),ALP含量、Runx2、OCN及OPN的mRNA表达、VEGF及BMP-2的蛋白表达均显著上调(均P＜0.05),处理后第5和7天,Runx2、OCN及OPN的mRNA表达显著上调(均P＜0.05),而常氧组差异无统计学意义(P＞0.05)。结论 低氧预处理的BMSCs对MC3T3-E1细胞的增殖和成骨分化具有促进作用。更多还原

【Abstract】 Objective To evaluate the effects of the serum-free supernatant from hypoxia-pretreated bone marrow mesenchymal stem cells(BMSCs) on the biological activity and osteogenic differentiation of mouse calvarial preosteoblasts(MC3T3-E1 cells).Methods MC3T3-E1 cells cultured with the serum-free supernatant from hypoxia-pretreated BMSCs served as the hypoxia group; those cultured with the serum-free supernatant from normoxia-cultured BMSCs served as the normoxia group; and cells cultured in regular medium without any supernatant addition served as the control group. On days 1, 3, 5, and 7 of culture, cell proliferation activity was assessed using the CCK-8 assay. Alkaline phosphatase(ALP) activity was detected with a biochemical kit. The mRNA expression levels of Runt-related transcription factor 2(Runx2), osteocalcin(OCN), and osteopontin(OPN) were measured by quantitative real-time polymerase chain reaction(qRT-PCR). Western blot was used to detect the protein expressions of vascular endothelial growth factor(VEGF) and bone morphogenetic protein 2(BMP-2). Results On days 1, 3, 5, and 7 after treatment, compared with the control group, the hypoxia group showed enhanced cell proliferation(P＜0. 05), significantly increased ALP activity, and upregulated mRNA expressions of Runx2, OCN, and OPN, as well as protein expressions of VEGF and BMP-2(all P＜0. 05). Notably, the mRNA expressions of Runx2, OCN, and OPN were more significantly upregulated on days 5 and 7(all P＜0. 05). In contrast, there were no statistically significant differences between the normoxia group and the control group(P＞0. 05). Conclusion The serum-free supernatant from hypoxia-pretreated BMSCs exerts a promotional effect on the proliferation and osteogenic differentiation of MC3T3-E1 cells.更多还原