钙离子对人牙囊细胞增殖、迁移和成骨分化的影响

doi:10.19439/j.sjos.2019.06.003

上海口腔医学 ›› 2019, Vol. 28 ›› Issue (6): 572-577.doi: 10.19439/j.sjos.2019.06.003

钙离子对人牙囊细胞增殖、迁移和成骨分化的影响

赵娴^1,2, 曾锦^1,2, 左东川³, 徐洁^1,2, 余京泓^1,2, 徐晓梅^1,2*

1.西南医科大学附属口腔医院正畸科,四川泸州 646000;
2.西南医科大学口颌面修复重建和再生实验室,
3.电生理学教育部重点实验室,心血管医学研究所,四川泸州 646000

收稿日期:2019-06-03 出版日期:2019-12-25 发布日期:2020-01-14
通讯作者: 徐晓梅,E-mail：xuxiaomei@hotmail.com
作者简介:赵娴(1993-),女,在读硕士研究生,E-mail：895572171@qq.com
基金资助:
泸州市-西南医科大学联合项目（2016LZXNYD-T04）; 泸州市科学技术和人才工作局应用基础研究（2018-JYJ-37）; 泸州市科技局-西南医科大学附属口腔医院联合项目（0800103009）

Effect of calcium on proliferation, migration and osteogenic differentiation of human dental follicle cells

ZHAO Xian^1,2, ZENG Jin^1,2, ZUO Dong-chuan³, XU Jie^1,2, YU Jing-hong^1,2, XU Xiao-mei^1,2

1. Department of Orthodontics, the Affiliated Stomatology Hospital of Southwest Medical University. Luzhou 646000;
2. Laboratory of Oral and Maxillofacial Reconstruction and Regeneration,
3.Institute of Cardiovascular Medicine, Key Laboratory of Electrophysiology of Ministry of Education, Southwest Medical University. Luzhou 646000, Sichuan Province, China

Received:2019-06-03 Online:2019-12-25 Published:2020-01-14

摘要/Abstract

摘要： 目的探讨Ca²⁺对人牙囊细胞增殖、迁移及成骨分化能力的影响。方法分离、培养人牙囊细胞（human dental follicle cells,hDFCs）,免疫荧光染色检测hDFCs来源,茜素红S和油红O染色鉴定多向分化能力;设置不同Ca²⁺浓度,CCK8法检测1、3、5、7 d时hDFCs的增殖能力;Transwell小室检测24 h时hDFCs的迁移能力;茜素红染色半定量分析法检测钙结节形成;反转录聚合酶链反应（RT-qPCR）检测成骨分化相关基因RUNT相关转录因子2（runt-related transcription factor 2,RUNX2）、骨钙素（osteocalcin,OCN）的表达。采用SPSS17.0软件包对数据进行统计学分析。结果与对照组相比,3、5、7 d时,3、4、5 mmol/L Ca²⁺显著促进hDFCs增殖（P<0.05）;3、4、5、6 mmol/L Ca²⁺显著促进hDFCs迁移（P<0.01）,高浓度Ca²⁺对其增殖、迁移无显著影响（P>0.05）。茜素红染色结果表明,Ca²⁺浓度达4 mmol/L时,可显著促进矿化结节生成（P<0.01）,矿化物生成与Ca²⁺呈浓度依赖性。RT-qPCR结果表明,Ca²⁺上调成骨分化相关基因RUNX2、OCN的表达（P<0.01）。结论低浓度Ca²⁺有利于人牙囊细胞增殖与迁移,而高浓度Ca²⁺更有利于人牙囊细胞成骨分化。

关键词: 人牙囊细胞, 钙离子, 迁移, 增殖, 成骨分化

Abstract: PURPOSE: To determine the role of Ca²⁺ in proliferation,migration and osteogenic differentiation of human dental follicle cells(hDFCs). METHODS: hDFCs were isolated and cultured. The source of hDFCs was detected by immunofluorescence staining. Osteogenesis and adipogenic differentiation of hDFCs was detected by alizarin red staining and oil red O staining, to identify its multi-directional differentiation ability. A series of Ca²⁺ solutions with different concentrations was prepared, CCK8 assay was used to detect the proliferative abilities at 1, 3, 5, and 7 d;migratory ability of 24 h was detected by Transwell assay. Calcium nodules were detected by semiquantitative analysis of alizarin red staining. mRNA expression of osteogenic differentiation related genes was examined by real-time quantitative polymerase chain reaction (RT-qPCR).Statistical analysis was performed using SPSS 17.0 software package. RESULTS: Compared with the control group, 3,4 and 5 mmol/L Ca²⁺ significantly promoted proliferation of hDFCs at 3, 5 and 7 d (P<0.05). 3, 4, 5 and 6 mmol/L Ca²⁺ significantly promoted the migration of hDFCs at 24 h(P<0.01). High concentration of Ca²⁺ had no significant effect on its proliferation and migration. The results of alizarin red staining showed that when Ca²⁺concentration reached 4 mmol/L, formation of mineralized nodules were increased(P<0.01), and Ca²⁺ concentration-dependent. RT-qPCR results showed that Ca²⁺ up-regulated the expression of RUNX2 and OCN in osteogenic differentiation genes (P<0.01). CONCLUSIONS: Low Ca²⁺ concentration is beneficial to proliferation and migration, and high Ca²⁺ concentration is beneficial to osteogenic differentiation of human dental follicle cells.

Key words: Human dental follicle cells, Calcium, Migration, Proliferation, Osteogenic differentiat

中图分类号:

R783.5

赵娴, 曾锦, 左东川, 徐洁, 余京泓, 徐晓梅. 钙离子对人牙囊细胞增殖、迁移和成骨分化的影响[J]. 上海口腔医学, 2019, 28(6): 572-577.

ZHAO Xian, ZENG Jin, ZUO Dong-chuan, XU Jie, YU Jing-hong, XU Xiao-mei. Effect of calcium on proliferation, migration and osteogenic differentiation of human dental follicle cells[J]. Shanghai Journal of Stomatology, 2019, 28(6): 572-577.

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钙离子对人牙囊细胞增殖、迁移和成骨分化的影响

Effect of calcium on proliferation, migration and osteogenic differentiation of human dental follicle cells

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