{"title":"氯化镁对人牙周韧带干细胞的成骨诱导活性研究。","authors":"Supanat Lumbikananda, Kittiphoj Tikkhanarak, Sarai Pongjantarasatian, Vorapat Trachoo, Worachat Namangkalakul, Thanaphum Osathanon","doi":"10.1016/j.identj.2024.11.013","DOIUrl":null,"url":null,"abstract":"<p><strong>Objectives: </strong>Periodontal ligament stem cells (PDLSCs) are promising for regenerative therapies due to their self-renewal and multilineage differentiation, essential for periodontal tissue repair. Although magnesium plays a vital role in bone metabolism, its specific effects on PDLSCs and potential applications in regeneration are unclear. This study aimed to investigate the effects of magnesium chloride (MgCl₂) on the proliferation and osteogenic differentiation of human PDLSCs (hPDLSCs).</p><p><strong>Methods: </strong>hPDLSCs were isolated, characterised, and treated with 0.1-40 mM MgCl₂. Cell viability and proliferation were assessed using an MTT assay. Cell migration was measured by a scratch assay. Colony-forming unit formation and cell cycle analysis were examined using crystal violet and propidium iodide staining. Osteogenic differentiation was assessed through alkaline phosphatase activity, Alizarin Red S staining, and RT-qPCR for osteogenic-related gene expression. RNA sequencing was performed to evaluate differential gene expression patterns in hPDLSCs treated with 10 mM MgCl₂. All statistical analyses were evaluated at P < .05.</p><p><strong>Results: </strong>hPDLSCs exhibited mesenchymal stem cell characteristics. MgCl₂ concentrations higher than 10 mM were cytotoxic. Significant increases in cell proliferation, colony-forming unit percentages, and active cell cycle activity were observed when treated with 0.1, 0.5, and 1 mM MgCl₂. However, MgCl₂ had no effect on cell migration. Mineralised nodule formation was observed in hPDLSCs treated with 0.1 and 0.5 mM MgCl₂ in osteogenic induction media, mediated by TRPM7 cation channel, along with upregulated expression of osteogenic marker genes. Bioinformatic analysis indicated alterations in chemokine signalling and cellular calcium homeostasis pathways when treated with 10 mM MgCl<sub>2</sub>.</p><p><strong>Conclusions: </strong>MgCl<sub>2</sub> at a dose of 0.1 mM is the most effective concentration to promote cell proliferation and stimulate osteogenic differentiation of hPDLSCs in vitro. These findings indicate that MgCl<sub>2</sub> enhances both the proliferation and osteogenic differentiation of hPDLSCs, supporting its potential application in periodontal tissues and alveolar bone regeneration.</p>","PeriodicalId":13785,"journal":{"name":"International dental journal","volume":" ","pages":"1431-1440"},"PeriodicalIF":3.2000,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Osteogenic Induction Activity of Magnesium Chloride on Human Periodontal Ligament Stem Cells.\",\"authors\":\"Supanat Lumbikananda, Kittiphoj Tikkhanarak, Sarai Pongjantarasatian, Vorapat Trachoo, Worachat Namangkalakul, Thanaphum Osathanon\",\"doi\":\"10.1016/j.identj.2024.11.013\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Objectives: </strong>Periodontal ligament stem cells (PDLSCs) are promising for regenerative therapies due to their self-renewal and multilineage differentiation, essential for periodontal tissue repair. Although magnesium plays a vital role in bone metabolism, its specific effects on PDLSCs and potential applications in regeneration are unclear. This study aimed to investigate the effects of magnesium chloride (MgCl₂) on the proliferation and osteogenic differentiation of human PDLSCs (hPDLSCs).</p><p><strong>Methods: </strong>hPDLSCs were isolated, characterised, and treated with 0.1-40 mM MgCl₂. Cell viability and proliferation were assessed using an MTT assay. Cell migration was measured by a scratch assay. Colony-forming unit formation and cell cycle analysis were examined using crystal violet and propidium iodide staining. Osteogenic differentiation was assessed through alkaline phosphatase activity, Alizarin Red S staining, and RT-qPCR for osteogenic-related gene expression. RNA sequencing was performed to evaluate differential gene expression patterns in hPDLSCs treated with 10 mM MgCl₂. All statistical analyses were evaluated at P < .05.</p><p><strong>Results: </strong>hPDLSCs exhibited mesenchymal stem cell characteristics. MgCl₂ concentrations higher than 10 mM were cytotoxic. Significant increases in cell proliferation, colony-forming unit percentages, and active cell cycle activity were observed when treated with 0.1, 0.5, and 1 mM MgCl₂. However, MgCl₂ had no effect on cell migration. Mineralised nodule formation was observed in hPDLSCs treated with 0.1 and 0.5 mM MgCl₂ in osteogenic induction media, mediated by TRPM7 cation channel, along with upregulated expression of osteogenic marker genes. Bioinformatic analysis indicated alterations in chemokine signalling and cellular calcium homeostasis pathways when treated with 10 mM MgCl<sub>2</sub>.</p><p><strong>Conclusions: </strong>MgCl<sub>2</sub> at a dose of 0.1 mM is the most effective concentration to promote cell proliferation and stimulate osteogenic differentiation of hPDLSCs in vitro. These findings indicate that MgCl<sub>2</sub> enhances both the proliferation and osteogenic differentiation of hPDLSCs, supporting its potential application in periodontal tissues and alveolar bone regeneration.</p>\",\"PeriodicalId\":13785,\"journal\":{\"name\":\"International dental journal\",\"volume\":\" \",\"pages\":\"1431-1440\"},\"PeriodicalIF\":3.2000,\"publicationDate\":\"2025-04-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"International dental journal\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1016/j.identj.2024.11.013\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2024/12/25 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q1\",\"JCRName\":\"DENTISTRY, ORAL SURGERY & MEDICINE\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"International dental journal","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1016/j.identj.2024.11.013","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/12/25 0:00:00","PubModel":"Epub","JCR":"Q1","JCRName":"DENTISTRY, ORAL SURGERY & MEDICINE","Score":null,"Total":0}
引用次数: 0
摘要
目的：牙周韧带干细胞（Periodontal ligament stem cells, PDLSCs）具有自我更新和多谱系分化的特性，在牙周组织修复中具有重要的应用前景。尽管镁在骨代谢中起着至关重要的作用，但其对PDLSCs的具体作用及其在再生中的潜在应用尚不清楚。本研究旨在探讨氯化镁（MgCl 2）对人PDLSCs （hPDLSCs）增殖和成骨分化的影响。方法：分离、表征hPDLSCs，用0.1 ~ 40 mM mgcl2处理。使用MTT法评估细胞活力和增殖。用划痕法测定细胞迁移量。结晶紫染色和碘化丙啶染色检测菌落形成单位形成和细胞周期分析。通过碱性磷酸酶活性、茜素红S染色和RT-qPCR检测成骨相关基因表达来评估成骨分化。通过RNA测序来评估10 mM MgCl 2处理hPDLSCs的差异基因表达模式。所有统计分析均以P < 0.05进行评价。结果：hPDLSCs表现出间充质干细胞的特征。浓度大于10 mM的MgCl 2具有细胞毒性。在0.1、0.5和1 mM MgCl₂处理下，细胞增殖、集落形成单位百分比和活跃的细胞周期活性显著增加。然而，MgCl 2对细胞迁移没有影响。在成骨诱导培养基中，用0.1和0.5 mM MgCl 2处理hPDLSCs，观察到矿化结节形成，由TRPM7阳离子通道介导，同时成骨标记基因表达上调。生物信息学分析表明，当10 mM MgCl2处理时，趋化因子信号传导和细胞钙稳态途径发生了变化。结论：0.1 mM剂量的MgCl2是体外促进hPDLSCs细胞增殖和促进成骨分化最有效的浓度。这些发现表明，MgCl2可以促进hPDLSCs的增殖和成骨分化，支持其在牙周组织和牙槽骨再生中的潜在应用。
Osteogenic Induction Activity of Magnesium Chloride on Human Periodontal Ligament Stem Cells.
Objectives: Periodontal ligament stem cells (PDLSCs) are promising for regenerative therapies due to their self-renewal and multilineage differentiation, essential for periodontal tissue repair. Although magnesium plays a vital role in bone metabolism, its specific effects on PDLSCs and potential applications in regeneration are unclear. This study aimed to investigate the effects of magnesium chloride (MgCl₂) on the proliferation and osteogenic differentiation of human PDLSCs (hPDLSCs).
Methods: hPDLSCs were isolated, characterised, and treated with 0.1-40 mM MgCl₂. Cell viability and proliferation were assessed using an MTT assay. Cell migration was measured by a scratch assay. Colony-forming unit formation and cell cycle analysis were examined using crystal violet and propidium iodide staining. Osteogenic differentiation was assessed through alkaline phosphatase activity, Alizarin Red S staining, and RT-qPCR for osteogenic-related gene expression. RNA sequencing was performed to evaluate differential gene expression patterns in hPDLSCs treated with 10 mM MgCl₂. All statistical analyses were evaluated at P < .05.
Results: hPDLSCs exhibited mesenchymal stem cell characteristics. MgCl₂ concentrations higher than 10 mM were cytotoxic. Significant increases in cell proliferation, colony-forming unit percentages, and active cell cycle activity were observed when treated with 0.1, 0.5, and 1 mM MgCl₂. However, MgCl₂ had no effect on cell migration. Mineralised nodule formation was observed in hPDLSCs treated with 0.1 and 0.5 mM MgCl₂ in osteogenic induction media, mediated by TRPM7 cation channel, along with upregulated expression of osteogenic marker genes. Bioinformatic analysis indicated alterations in chemokine signalling and cellular calcium homeostasis pathways when treated with 10 mM MgCl2.
Conclusions: MgCl2 at a dose of 0.1 mM is the most effective concentration to promote cell proliferation and stimulate osteogenic differentiation of hPDLSCs in vitro. These findings indicate that MgCl2 enhances both the proliferation and osteogenic differentiation of hPDLSCs, supporting its potential application in periodontal tissues and alveolar bone regeneration.
期刊介绍：
The International Dental Journal features peer-reviewed, scientific articles relevant to international oral health issues, as well as practical, informative articles aimed at clinicians.
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