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目的:探究不同浓度Mg~(2+)对hPDLCs成骨向分化的影响,为后续牙周组织再生实验选择合适的Mg~(2+)注入浓度提供依据。方法:取P3-P5代hPDLCs于Mg~(2+)浓度分别为0、10、15、25、35、50mmol/L条件中常规培养,用CCK-8法检测hPDLCs增殖情况。成骨诱导后比较各组ALP活性差异及茜素红矿化结节着色情况,进行RT-qPCR检测成骨相关基因Runx2、ALP、Col1、OPN及Bglap的表达差异。采用SPSS16.0软件,运用单因素方差分析进行统计学分析。结果:10、15、25 mmol/L Mg~(2+)浓度可促进细胞增殖并提高ALP活性,35、50 mmol/L Mg~(2+)浓度抑制细胞增殖及ALP活性。结论:适宜浓度镁离子(0~25mmol/L)可促进hPDLCs早期成骨分化并抑制矿化。
OBJECTIVE: To investigate the effects of different concentrations of Mg 2+ on the osteogenic differentiation of hPDLCs, and to provide a basis for selecting suitable concentrations of Mg 2+ for periodontal regeneration experiments. Methods: P3-P5 generation hPDLCs were cultured routinely in Mg 2+ concentrations of 0, 10, 15, 25, 35 and 50 mmol / L respectively. Proliferation of hPDLCs was detected by CCK-8 assay. After osteogenic induction, the differences of ALP activity and the alizarin red mineralization nodules were compared. The expressions of osteoblast-related gene Runx2, ALP, Col1, OPN and Bglap were detected by RT-qPCR. Using SPSS16.0 software, using one-way analysis of variance analysis. Results: 10, 15, 25 mmol / L Mg 2+ concentration could promote cell proliferation and ALP activity. 35,50 mmol / L Mg 2+ concentration could inhibit cell proliferation and ALP activity. Conclusion: Appropriate concentration of magnesium ion (0 ~ 25mmol / L) can promote the early osteogenic differentiation and inhibit the mineralization of hPDLCs.