镁离子影响人上颌窦黏膜干细胞体外成骨分化的研究

Effect of Magnesium Ions on Biological Activity of Human Maxillary Sinus Membrane Stem Cells

目的:研究不同浓度镁离子(Mg^(2+))对人上颌窦黏膜干细胞(human maxillary sinus membrane stem cells,hMSMSCs)生物行为的影响。方法:体外提取hMSMSCs后利用流式细胞术和多向分化实验来鉴定其间充质干细胞特征,随后将细胞分别培养在0.8 mmol/L、1.8 mmol/L、2.8 mmol/L和5.8 mmol/L的Mg^(2+)浓度下。通过CCK-8实验和Calcein AM/PI染色检测hMSMSCs的细胞黏附和增殖,采用罗丹明鬼笔环肽-DAPI染色评估细胞的黏附伸展。对成骨诱导培养后的hMSMSCs进行ALP活性、ALP染色和茜素红染色,同时利用qRT-PCR实验检测成骨相关生长因子的表达差异。结果:适宜浓度Mg^(2+)能有效改善hMSMSCs的黏附、伸展和增殖。1.8 mmol/L Mg^(2+)提高了hMSMSCs的ALP活性,并显著改善细胞外钙结节的形成并上调成骨相关基因的表达,而过高浓度的Mg^(2+)相对抑制了hMSMSCs的成骨分化(P<0.05)。结论:适宜浓度的Mg^(2+)(1.8 mmol/L)可以促进hMSMSCs的细胞骨架的伸展和黏附,并有利于增殖和成骨分化。

Objective:To investigate the effects of different concentrations of magnesium ions(Mg^(2+))on the biological behavior of human maxillary sinus membrane stem cells(hMSMSCs).Methods:hMSMSCs were extracted in vitro,and flow cytometry and multilineage differentiation experiments were conducted to identify their mesenchymal stem cell characteristics.Subsequently,the cells were cultured at concentrations of 0.8 mmol/L,1.8 mmol/L,2.8 mmol/L,and 5.8 mmol/L of Mg^(2+).Cell adhesion and proliferation of hMSMSCs were evaluated using CCK-8 assay and Calcein AM/PI staining,while cell adhesion extension was assessed using Rhodamine phalloidin-DAPI staining.ALP activity,ALP staining,and Alizarin Red staining were performed on hMSMSCs after osteogenic induction culture,and qRT-PCR experiments were conducted to detect the expression differences of osteogenic-related growth factors.Results:Optimal concentration of Mg^(2+)effectively improved cell adhesion,extension,and proliferation of hMSMSCs.1.8 mmol/L Mg^(2+)enhanced ALP activity in hMSMSCs,significantly improved extracellular calcium nodule formation,and upregulated the expression of osteogenic-related genes.Conversely,excessively high Mg^(2+)concentrations inhibited osteogenic differentiation of hMSMSCs(P<0.05).Conclusion:Appropriate concentration of Mg^(2+)(1.8 mmol/L)promotes cell spreading and adhesion of hMSMSCs,facilitating proliferation and osteogenic differentiation.