Zhang Zhihao, Gong Ning, Wang Ying, Xu Lei, Zhao Sinan, Liu Yanshan, Tan Fei
Department of Prosthodontic, The Affiliated Hospital of Qingdao University, Qingdao, 266003, China.
School of Stomatology, Qingdao University, Qingdao, 266023, China.
Biol Trace Elem Res. 2025 Apr;203(4):1922-1933. doi: 10.1007/s12011-024-04303-4. Epub 2024 Aug 16.
Human Maxillary Sinus Membrane Stem Cells (hMSMSCs) contribute significantly to bone formation following maxillary sinus floor augmentation (MSFA). The biological behavior of mesenchymal stem cells is notably influenced by varying concentrations of magnesium (Mg), strontium (Sr), and zinc (Zn) ions; however, their specific effects on hMSMSCs have not been comprehensively studied. We isolated hMSMSCs and identified their mesenchymal stem cell characteristics by flow cytometry and multilineage differentiation experiments. Subsequently, the hMSMSCs were cultured in media containing different concentrations of these metal ions. The proliferation and viability of hMSMSCs were assessed using CCK-8 and Calcein AM/PI staining. After osteogenic induction, cells were evaluated for alkaline phosphatase (ALP) activity, ALP staining, and Alizarin Red staining. Additionally, qRT-PCR was used to detect differences in osteogenic gene expression, and immunofluorescence staining was used to observe variations in OCN protein levels. The results indicated that 1 mM Mg, 0.01 mM Sr, and 0.001 mM Zn significantly improved the proliferation and activity of hMSMSCs. These concentrations also notably enhanced ALP secretion, increased bone-related gene expression, and augmented osteocalcin expression and formation of extracellular calcium nodules, thereby improving osteogenic differentiation. However, higher concentrations of Mg, Sr, and Zn decreased cell viability and osteogenic differentiation. Mg, Sr, and Zn promote osteogenic differentiation and proliferation of hMSMSCs in a concentration-dependent manner, indicating that the type and concentration of ions in the extracellular environment can significantly alter hMSMSCs behavior, which is a crucial consideration for material design in maxillary sinus elevation applications.
人上颌窦膜干细胞（hMSMSCs）在上颌窦底提升术（MSFA）后对骨形成有显著贡献。间充质干细胞的生物学行为受到不同浓度的镁（Mg）、锶（Sr）和锌（Zn）离子的显著影响；然而，它们对hMSMSCs的具体影响尚未得到全面研究。我们分离出hMSMSCs，并通过流式细胞术和多谱系分化实验鉴定其间充质干细胞特征。随后，将hMSMSCs培养在含有不同浓度这些金属离子的培养基中。使用CCK-8和钙黄绿素AM/PI染色评估hMSMSCs的增殖和活力。在进行成骨诱导后，评估细胞的碱性磷酸酶（ALP）活性、ALP染色和茜素红染色。此外，使用qRT-PCR检测成骨基因表达的差异，并使用免疫荧光染色观察骨钙素（OCN）蛋白水平的变化。结果表明，1 mM Mg、0.01 mM Sr和0.001 mM Zn显著改善了hMSMSCs的增殖和活性。这些浓度还显著增强了ALP分泌，增加了骨相关基因的表达，并增强了骨钙素的表达和细胞外钙结节的形成，从而改善了成骨分化。然而，更高浓度的Mg、Sr和Zn会降低细胞活力和成骨分化。Mg、Sr和Zn以浓度依赖的方式促进hMSMSCs的成骨分化和增殖，表明细胞外环境中离子的类型和浓度可显著改变hMSMSCs的行为，这是上颌窦提升应用中材料设计的关键考虑因素。
