高浓度唑来膦酸对人牙周膜干细胞凋亡及成骨分化影响的实验研究

摘要/Abstract

摘要： 目的：研究高浓度唑来膦酸对体外分离的健康人牙周膜干细胞增殖、凋亡和成骨分化的影响。方法：体外分离培养健康人牙周膜干细胞，用不同浓度的唑来膦酸（5μmol/L、10 μmol/L）处理，不加药组为空白对照。以 CCK8 检测细胞增殖，流式细胞技术检测细胞凋亡，ALP、茜素红染色及半定量分析检测细胞体外成骨分化，荧光定量PCR检测成?标志物I型胶原(COL1A1)和骨钙素（OCN）基因的表达，免疫荧光检测 OCN和 COL1A1 蛋白的表达。结果：唑来膦酸可呈计量依赖性抑制人牙周膜干细胞增殖；唑来膦酸用药组细胞的凋亡率显著?于对照组，且随着药物浓度的增加，凋亡百分比逐渐上升；ALP和茜素红染?结果表示唑来膦酸用药组细胞的体外成骨分化能力较对照组显著下降；荧光定量PCR和免疫荧光结果提示，药物处理后OCN和 COL1A1的基因表达和蛋白表达均降低。结论：高浓度唑来膦酸显著抑制人牙周膜干细胞的增殖及体内外成骨分化，并诱导其凋亡。

关键词: 唑来膦酸, 人牙周膜干细胞, 颌骨坏死, 凋亡, 成骨分化

Abstract: Objective To investigate the effect of high concentration of zoledronate on proliferation, apoptosis and osteogenic differentiation of healthy human periodontal stem cells. Methods Human periodontal ligament stem cells （hPDLSCs）were isolated from health tissues and exposed to zoledronate with different concentrations (0 μmol/L, 5μmol/L，10 μmol/L) in vitro. Cell proliferation ability was detected using CCK-8 kit and flow cytometry was applied to identity cell apoptosis. ALP staining and Alizarin red staining were used to evaluate the osteogenesis differentiation of hPDLSCs. Real-time PCR were used to measure the gene expression of COL1A1 and OCN and immunofluorescence was used to detect the levels of protein expression of COL1A1 and OCN. Osteogenesis was analyzed with ectopic osteogenesis in nude mice. Results Zoledronate inhibited proliferation of hPDLSCs in a dose dependent manner. Flow cytometry results indicated higher proportion of apoptosis cells in zoledronate treated groups. ALP staining and Alizarin red staining both revealed decreased osteogenesis differentiation ability in zoledronate treated groups. Significant down-regulation of the levels of mRNA expression and protein expression in COL1A1 and OCN were also found in zoledronate treated groups. Conclusion High concentration of zoledronate significantly inhibited the proliferation and osteogenic differentiation of hPDLSCs and induced their apoptosis.

Key words: zoledronate, human periodontal ligament stem cell (hPDLSCs), Osteonecrosis of the jaw, Apoptosis, Osteogenesis differentiation

李萌宇俞叶佳施越琦戈旌王绍义. 高浓度唑来膦酸对人牙周膜干细胞凋亡及成骨分化影响的实验研究[J]. , 2019, 39(10): 880-888.

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