中国组织工程研究

中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (19): 3005-3010.doi: 10.12307/2023.639

• 牙髓及牙周膜干细胞 Dental pulp and periodontal ligament stem cells • 上一篇    下一篇

不同浓度钙离子干预人牙周膜干细胞的增殖和成骨分化

李颖辉1，2，齐芳芳2，3，韩  行1，2，赵力如1，2，马文盛1，2   

  1. 1河北医科大学口腔医院正畸科，河北省石家庄市   050000；2河北省口腔医学重点实验室，河北省石家庄市   050000；3河北医科大学口腔医院牙体牙髓科，河北省石家庄市   050000
  • 收稿日期:2022-06-14 接受日期:2022-08-19 出版日期:2023-07-08 发布日期:2022-11-28
  • 通讯作者: 马文盛，博士，教授，河北医科大学口腔医院正畸科，河北省口腔医学重点实验室，河北省石家庄市 050000
  • 作者简介:李颖辉，男，1983年生，河北省任丘市人，汉族，2017年日本信州大学毕业，博士，讲师，主要从事口腔正畸临床与基础方面的研究。
  • 基金资助:
    河北省卫生健康委医学科学研究课题计划(20191077)，项目负责人：李颖辉；河北省人社厅引进留学人员资助项目(C201858)，项目负责人：李颖辉

Different concentrations of calcium ions interfere with the proliferation and osteogenic differentiation of human periodontal ligament stem cells

Li Yinghui1, 2 , Qi Fangfang2, 3, Han Xing1, 2 , Zhao Liru1, 2 , Ma Wensheng1, 2   

  1. 1Department of Orthodontics, Hospital of Stomatology, Hebei Medical University, Shijiazhuang 050000, Hebei Province, China; 2Hebei Key Laboratory of Stomatology, Shijiazhuang 050000, Hebei Province, China; 3Department of Operative Dentistry and Endodontics, Hospital of Stomatology, Hebei Medical University, Shijiazhuang 050000, Hebei Province, China
  • Received:2022-06-14 Accepted:2022-08-19 Online:2023-07-08 Published:2022-11-28
  • Contact: Ma Wensheng, PhD, Professor, Department of Orthodontics, Hospital of Stomatology, Hebei Medical University, Shijiazhuang 050000, Hebei Province, China; Hebei Key Laboratory of Stomatology, Shijiazhuang 050000, Hebei Province, China
  • About author:Li Yinghui, PhD, Lecturer, Department of Orthodontics, Hospital of Stomatology, Hebei Medical University, Shijiazhuang 050000, Hebei Province, China; Hebei Key Laboratory of Stomatology, Shijiazhuang 050000, Hebei Province, China
  • Supported by:
    the Medical Science Research Project of Hebei Provincial Health Commission, No. 20191077 (to LYH); the Hebei Provincial Department of Human Resources and Social Security Introduced Overseas Students Funding Project, No. C201858 (to LYH)

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摘要：


文题释义：

牙周膜干细胞：是存在于牙周膜中的一种未分化的间充质干细胞，具有高增殖能力和多向分化能力，能定向分化为成骨细胞及成脂细胞，其分化能力受细胞因子、激素、药物等诸多因素的调节。
钙离子(Ca2+)：机体各项生理活动不可缺少的离子，其对于维持细胞膜两侧的生物电位、正常的神经传导功能、正常的肌肉伸缩与舒张功能以及神经-肌肉传导功能有重要作用，还有一些激素的作用机制均通过钙离子表现出来。

背景：钙离子作为第二信使，调控着众多的生理过程，但在牙周膜干细胞成骨分化方面的生物学效应尚不明确。
目的：探讨钙离子在人牙周膜干细胞增殖和成骨分化过程中的作用。
方法：使用含钙离子浓度为0(对照组)，1，2，5，10，15 mmol/L的完全培养基及成骨诱导液对第3代人牙周膜干细胞进行增殖能力检测及成骨诱导。培养1，3，5，7 d使用CCK-8检测细胞增殖能力；在成骨诱导第3，7天采用Real-Time PCR和Western blot检测RUNX2、OPN、OCN的基因及蛋白表达；成骨诱导第14天时进行茜素红染色分析人牙周膜干细胞的成骨矿化情况。
结果与结论：①CCK-8检测结果显示，2，5 mmol/L钙离子组人牙周膜干细胞在第5，7天的增殖能力优于对照组(P < 0.05)；②体外成骨诱导7 d后，5，10 mmol/L钙离子组人牙周膜干细胞中RUNX2、OPN、OCN的表达高于对照组(P < 0.05)；③体外成骨诱导14 d后，含钙离子组人牙周膜干细胞的矿化结节数量均高于对照组(P < 0.05)；④以上结果表明，在一定浓度范围内，钙离子可促进人牙周膜干细胞的增殖与成骨分化。
https://orcid.org/0000-0002-8813-3961 (李颖辉) 
中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 钙离子, 人牙周膜干细胞, 增殖, 成骨分化

Abstract: BACKGROUND: As a second messenger, Ca2+ regulates many physiological processes, but its biological effect on the osteogenic differentiation of periodontal ligament stem cells is still unclear.  
OBJECTIVE: To investigate the effect of Ca2+ on the osteogenic differentiation of human periodontal ligament stem cells.
METHODS: The proliferation ability and osteogenic differentiation of the third generation human periodontal ligament stem cells were detected by the complete medium containing Ca2+ at the concentrations of 0 (control group), 1, 2, 5, 10, and 15 mmol/L and osteogenic induction solution.  CCK-8 assay was used to detect cell proliferation at 1, 3, 5, and 7 days. Real-time PCR and western blot assay were used to detect the expression levels of genes and proteins related to osteogenic differentiation factors RUNX2, OPN and OCN on days 3 and 7 of osteogenic induction. Alizarin red staining was performed on day 14 to analyze the osteogenic mineralization of human periodontal ligament stem cells.  
RESULTS AND CONCLUSION: (1) CCK-8 assay showed that the proliferation ability of human periodontal ligament stem cells in 2 and 5 mmol/L groups was better than that in control group on days 5 and day 7 (P < 0.05).  (2) On day 7 after in vitro osteogenic induction, the expression levels of osteogenic genes RUNX2, OPN and OCN in human periodontal ligament stem cells of 5 and 10 mmol/L groups were higher than those in control group (P < 0.05). (3) After 14 days of in vitro osteogenic induction, the numbers of mineralization nodules of human periodontal ligament stem cells in Ca2+ groups were higher than that in control group (P < 0.05). (4) These results suggest that Ca2+ can promote the proliferation and osteogenic differentiation of human periodontal ligament stem cells in a certain concentration range.

Key words: Ca2+, human periodontal ligament stem cell, proliferation, osteogenic differentiation

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