上海口腔医学

上海口腔医学 ›› 2025, Vol. 34 ›› Issue (5): 465-471.doi: 10.19439/j.sjos.2025.05.003

• 论著 • 上一篇    下一篇

掺锂化合物诱导乳牙牙髓干细胞成牙本质分化的实验研究

黄文燕1, 黄宇航1, 何雁冰2, 高敏1, 钟宁1, 吕锦鸿1, 李子凌1, 李馨1, 曾素娟1   

  1. 1.广州医科大学口腔医学院·附属口腔医院儿童 口腔科,广东省口腔组织修复与重建工程技术研究中心, 广州市口腔再生医学基础与应用研究重点实验室,广东 广州 510182;
    2.江门市中心医院 口腔科,广东 江门 529000
  • 收稿日期:2024-05-31 修回日期:2024-07-03 出版日期:2025-10-25 发布日期:2025-10-31
  • 通讯作者: 曾素娟,E-mail：Zengsujuan78@foxmail.com
  • 作者简介:黄文燕（1992-）,女,硕士,主治医师,E-mail：1545024244@qq.com
  • 基金资助:
    广州市卫生健康科技项目一般引导项目（20231A011105）

Experimental study of odontoblast differentiation of stem cells from human exfoliated deciduous teeth induced by lithium-doped compounds

Huang Wenyan1, Huang Yuhang1, He Yanbing2, Gao Min1, Zhong Ning1, Lyu Jinhong1, Li Ziling1, Li Xin1, Zeng Sujuan1   

  1. 1. Department of Pediatric Dentistry, School and Hospital of Stomatology, Guangzhou Medical University; Guangdong Engineering Research Center of Oral Restoration and Reconstruction; Guangzhou Key Laboratory of Basic and Applied Research of Oral Regenerative Medicine. Guangzhou 510182;
    2. Department of Stomatology, Jiangmen Central Hospital. Jiangmen 529000, Guangdong Province, China
  • Received:2024-05-31 Revised:2024-07-03 Online:2025-10-25 Published:2025-10-31

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摘要： 目的:以添加氯化锂（LiCl）的矿化液模拟成牙本质分化环境,探讨其诱导乳牙牙髓干细胞（stem cells from human exfoliated deciduous teeth,SHEDs）成牙本质分化的潜能。方法:提取原代SHEDs进行培养和鉴定,取P3~6 SHEDs进行后续实验。配制含0、1.0、2.5、5.0、10 mmol/L LiCl的成骨培养基（osteogenic medium,OM）用于诱导SHEDs成牙本质分化,1、3、5、7天时以CCK-8法检测细胞增殖能力。4、7、10天时以碱性磷酸酶染色检测细胞ALP活性。10天时,以RT-PCR检测DSPP、COL-1、Runx2基因水平,Western免疫印记法检测DSPP、Runx2蛋白水平。14天时,以茜素红染色和钙化结节半定量检测细胞矿化能力,筛选出最佳LiCl浓度。结果:CCK-8结果显示,低浓度（1.0、2.5 mmol/L）LiCl基本不影响细胞增殖,高浓度（5.0、10 mmol/L）LiCl抑制细胞增殖。4、7、10天时,低浓度LiCl提高SHEDs的ALP活性,其中,2.5 mmol/L LiCl组效果最佳,而高浓度LiCl则抑制ALP活性。RT-PCR及Western免疫印迹结果显示,低浓度LiCl促进SHEDs内DSPP、COL-1的基因表达及DSPP、Runx2蛋白表达。茜素红染色结果显示,1.0、2.5、5.0 mmol/L LiCl提高SHEDs矿化能力,10 mmol/L LiCl组与对照组形成的矿化结节量无显著差异。结论:低浓度锂化合物可促进SHEDs向成牙本质分化。

关键词: 人乳牙牙髓干细胞, 氯化锂, 成牙本质分化, 矿化能力

Abstract: PURPOSE: To simulate the mineralizing fluid with lithium chloride (LiCl) as a dentinogenic environment and investigate its potential to induce stem cells from human exfoliated deciduous teeth (SHEDs) into dentinogenesis. METHODS: Primary SHEDs were isolated, cultured, and identified, and P3-6 SHEDs were used for subsequent experiments. Osteogenic medium(OM) containing 0, 1.0, 2.5, 5.0, 10 mmol/L LiCl were prepared. Cell proliferation was assessed using CCK-8 assay at 1, 3, 5, and 7 days. Alkaline phosphatase(ALP) activity was examined by staining at 4, 7, and 10 days. Gene expression levels of DSPP, Runx2 and COL-1 were detected by RT-PCR, and protein levels of DSPP and Runx2 were analyzed by Western blot at 10 days. Alizarin red staining and semi-quantitative analysis were conducted at 14 days to evaluate mineralization capacity and select the optimal LiCl concentration. RESULTS: CCK-8 assay showed that low concentrations(≤2.5 mmol/L) of LiCl had minimal effect on cell proliferation from 1 to 7 days, while high concentrations (>2.5 mmol/L) inhibited cell proliferation. ALP activity was enhanced by low concentrations of LiCl at 4, 7, and 10 days, with the 2.5 mmol/L LiCl group showing the most significant effect, while high concentrations suppressed ALP activity. Western blot results revealed that low concentrations of LiCl promoted the protein expression of DSPP and Runx2 in SHEDs. RT-PCR demonstrated that low concentrations of LiCl upregulated the gene expression of DSPP and COL-1 in SHEDs. Qualitative and quantitative Alizarin red staining indicated that 1.0, 2.5, 5.0 mmol/L LiCl improved SHEDs mineralization capacity, while 10 mmol/L LiCl showed no significant difference in mineralized nodule formation compared to the control group. CONCLUSIONS: Low concentrations of lithium compounds can promote the dentinogenesis of SHEDs.

Key words: Stem cells from human exfoliated deciduous teeth, Lithium chloride, Dentinogenesis, Mineralization capacity

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