氟暴露对人牙周膜干细胞增殖及成骨分化的影响

doi:10.13701/j.cnki.kqyxyj.2020.09.015

口腔医学研究 ›› 2020, Vol. 36 ›› Issue (9): 866-870.DOI: 10.13701/j.cnki.kqyxyj.2020.09.015

氟暴露对人牙周膜干细胞增殖及成骨分化的影响

邱吟枫¹, 汤颖¹, 沈忆芬¹, 刘超¹, 沈昊¹, 顾永春^1*, 于金华^2*

1.苏州市第九人民医院口腔科江苏苏州 215200;
2.江苏省口腔疾病研究重点实验室江苏南京 210029

收稿日期:2020-02-22 出版日期:2020-09-28 发布日期:2020-09-15
通讯作者: *顾永春,E-mail: guyc7152@163.com;于金华,E-mail: yujinhua@njmu.edu.cn
作者简介:邱吟枫(1989~),男,江苏苏州人,学士,住院医师,主要从事口腔临床医学工作。
基金资助:
江苏省口腔疾病研究重点实验室开放课题基金(编号:JSK-LOD-KF-1705);苏州市吴江区卫健委科教兴卫项目(编号:wwk201705)

Effects of Fluoride Exposure on Cell Proliferation and Osteogenic Differentiation of Periodontal Ligament Stem Cells

QIU Yinfeng¹, TANG Ying¹, SHEN Yifen¹, LIU Chao¹, SHEN Hao¹, GU Yongchun^1*, YU Jinhua^2*

1. Ninth People's Hospital of Suzhou, Suzhou 215200, China;
2. Jiangsu Key Laboratory of Oral Diseases, Nanjing 210029, China

Received:2020-02-22 Online:2020-09-28 Published:2020-09-15

摘要/Abstract

摘要： 目的: 探讨氟暴露对牙周膜干细胞(periodontal ligment stem cells, PDLSCs)增殖及成骨分化的影响。方法: 从人牙周膜组织中分离、培养得到PDLSCs,取第3代细胞行流式细胞分析干细胞表面标志物。分别用含0.1、1、10、20、40 mg/L的含氟培养基进行培养,检测不同浓度氟对PDLSCs细胞增殖(CCK-8法)、集落形成、细胞周期变化(流式细胞分析)、成骨分化能力(茜素红染色及Western blot分析ALP及RUNX2的蛋白表达水平)的影响。结果: 氟暴露对PDLSCs增殖与成骨分化的影响呈浓度及时间依赖性。0.1~10 mg/L氟对PDLSCs增殖无显著影响(P>0.05),但0.1及1 mg/L氟可显著促进其成骨分化,表现为钙化结节增多(P<0.05),ALP及RUNX2蛋白表达水平升高(P<0.05)。高浓度氟(20、40 mg/L)显著抑制PDLSCs增殖(P<0.05),具有较强的细胞毒性。结论: 氟暴露会影响PDLSCs的增殖与成骨分化。高浓度氟抑制PDLSCs增殖,而氟浓度较低时(0.1、1 mg/L)对PDLSCs增殖无显著影响,并可以促进细胞成骨分化。

关键词: 牙周膜间充质干细胞, 氟暴露, 细胞增殖, 细胞分化

Abstract: Objective: To disclose the biological effects of fluoride (F) exposure on cell proliferation and osteogenic differentiation of periodontal ligament stem cells (PDLSCs). Methods: PDLSCs were isolated and cultured from human periodontal ligament tissues. The surface markers of passage 3 cells were analyzed by flow cytometry. After treated with different concentrations of F (0.1-40 mg/L) for indicated period of time, the effects of F exposure on cell proliferation (CCK-8 assay), capability of colony-forming, cell circle phase changes (flow cytometry), as well as the osteogenic potential (Alizarin red staining and Western blot assay of expression of RUNX2 and ALP) were examined. Results: F exposure affected the cell proliferation and osteogenic differentiation of PDLSCs in a dose- and time-dependent manner. 0.1-10 mg/L F did not significantly affect cell proliferation (P>0.05), while 0.1 and 1 mg/L F significantly enhanced the osteogenic potential, which manifested as increased calcified nodules and upregualtion of RUNX2 and ALP expression (P<0.05). High dose of F (20 and 40 mg/L) significantly inhibited the cell proliferation (P<0.05), exhibiting remarkable cytotoxicity. Conclusion: F exposure affects the proliferation and osteogenic differentiation of PDLSCs. High concentration of F inhibits the proliferation of PDLSCs, while low F concentration (0.1 and 1 mg/L) has no significant effect on the proliferation of PDLSCs, and can promote cell osteogenic differentiation.

Key words: periodontal ligament stem cells, fluoride exposure, cell proliferation, cell differentiation

邱吟枫, 汤颖, 沈忆芬, 刘超, 沈昊, 顾永春, 于金华. 氟暴露对人牙周膜干细胞增殖及成骨分化的影响[J]. 口腔医学研究, 2020, 36(9): 866-870.

QIU Yinfeng, TANG Ying, SHEN Yifen, LIU Chao, SHEN Hao, GU Yongchun, YU Jinhua. Effects of Fluoride Exposure on Cell Proliferation and Osteogenic Differentiation of Periodontal Ligament Stem Cells[J]. Journal of Oral Science Research, 2020, 36(9): 866-870.

参考文献

[1] Barbier O, Arreola-Mendoza L, Del Razo LM. Molecular mechanisms of fluoride toxicity [J]. Chem Biol Interact, 2010,188(2):319-333.
[2] Li X, Meng L, Wang F, et al. Sodium fluoride induces apoptosis and autophagy via the endoplasmic reticulum stress pathway in MC3T3-E1 osteoblastic cells [J]. Mol Cell Biochem, 2019, 454(1-2):77-85.
[3] Aulestia FJ, Groeling J, Bomfim GHS, et al. Fluoride exposure alters Ca²⁺ signaling and mitochondrial function in enamel cells [J]. Sci Signal, 2020, 13(619): eaay0086.
[4] Wang L, Zhu Y, Wang D. High-dose fluoride induces apoptosis and inhibits ameloblastin secretion in primary rat ameloblast [J]. Biol Trace Element Res, 2016, 174(2):402-409.
[5] Wu RX, Bi CS, Yu Y, et al. Age-related decline in the matrix contents and functional properties of human periodontal ligament stem cell sheets [J]. Acta Biomater, 2015, 22:70-82.
[6] 郑树灿,邹晖,黄旭瑶,等.低氧对人牙周膜干细胞增殖和分化的影响[J].口腔医学研究, 2019, 35(1):43-46.
[7] 吴晓玲,郑茜,吕佳岭,等.雌激素微环境下非经典Wnt/Ca²⁺信号通路调控人牙周膜干细胞成骨分化的研究[J].口腔医学研究,2019,35(1):38-42.
[8] Seo BM, Miura M, Gronthos S, et al. Investigation of multipotent postnatal stem cells from human periodontal ligament [J]. Lancet, 2004, 364(9429):149-155.
[9] Jin SH, Lee JE, Yun JH, et al. Isolation and characterization of human mesenchymal stem cells from gingival connective tissue [J]. J Periodontal Res, 2015, 50(4):461-467.
[10] Zhu W, Liang M. Periodontal ligament stem cells: current status, concerns, and future prospects [J]. Stem Cells Int, 2015, 2015:972313.
[11] Li KQ, Jia SS, Ma M, et al. Effects of fluoride on proliferation and mineralization in periodontal ligament cells in vitro [J]. Braz J Med Biol Res, 2016, 49(8): e5291.
[12] Yang C, Wang Y, Xu H. Fluoride regulate osteoblastic transforming growth factor-β1 Signaling by mediating recycling of the type I receptor ALK5 [J]. PLoS One, 2017, 12(1): e0170674.
[13] Jain RB. Concentrations of fluoride in water and plasma for US children and adolescents: Data from NHANES 2013-2014 [J]. Environ Toxicol Phar, 2017, 50:20-31.
[14] 李海燕.饮水氟含量及尿氟、血清氟与氟斑牙患病率的相关性研究[J].中国卫生工程学,2017,16(4):512-513.
[15] Singer L, Ophaug R. Ionic and nonionic fluoride in plasma (or serum) [J]. Crit Rev Clin Lab Sci, 1982, 18(2): 111-140.
[16] Yi J, Weir MD, Melo MAS, et al. Novel rechargeable nano-CaF2 orthodontic cement with high levels of long-term fluoride release [J]. J Dent, 2019, 90:103214.
[17] Qiao W, Liu R, Li Z, et al. Contribution of the in situ release of endogenous cations from xenograft bone driven by fluoride incorporation toward enhanced bone regeneration [J]. Biomater Sci, 2018, 6(11): 2951-2964.

氟暴露对人牙周膜干细胞增殖及成骨分化的影响

Effects of Fluoride Exposure on Cell Proliferation and Osteogenic Differentiation of Periodontal Ligament Stem Cells

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

[1]	张师贤, 林成泉, 谢长富, 陈伟辉. Gli1抑制剂GANT61对小鼠牙胚体外发育的影响[J]. 口腔医学研究, 2021, 37(1): 23-27.
[2]	尚佳鑫, 苏瑞, 马冲, 李新月. 两种抛光方式对两种充填材料表面形貌和细胞增殖能力的影响[J]. 口腔医学研究, 2020, 36(6): 581-584.
[3]	王薛藤, 聂敏海, 左东川, 邓浩, 曾锦. TRPM4通道对人口腔颊黏膜成纤维细胞增殖和迁移的影响[J]. 口腔医学研究, 2020, 36(5): 490-495.
[4]	张维甲, 潘爽, 李艳萍, 何丽娜, 吴作栋, 牛玉梅. 兔牙髓干细胞与雪旺细胞体外间接共培养对增殖和兔牙髓干细胞成神经向分化的研究[J]. 口腔医学研究, 2020, 36(10): 968-972.
[5]	黄奕智, 郭俊, 杨健. 富血小板纤维蛋白对牙周膜成纤维细胞成骨能力影响的实验研究[J]. 口腔医学研究, 2020, 36(1): 41-45.
[6]	于淼, 李倜, 白建文, 王丽美, 段晓琪, 孙龙, 孙钦峰. ProgranuIin衍生工程蛋白Atsttrin对牙周膜干细胞成骨分化作用的影响[J]. 口腔医学研究, 2019, 35(7): 643-646.
[7]	程海燕, 沈兰花, 张瑞, 孟玲娜, 刘迪, 刑北昱, 陶冠男. 雌孕激素对人牙周膜细胞成骨分化影响的研究[J]. 口腔医学研究, 2019, 35(7): 657-660.
[8]	徐方方,殷丽华. 富血小板纤维蛋白对干细胞增殖及成骨分化的研究进展[J]. 口腔医学研究, 2019, 35(6): 527-530.
[9]	肖婉鲁,潘爽,侯婷婷,李艳萍,何丽娜,牛玉梅. 载柚皮苷胶原蛋白凝胶对人牙髓干细胞增殖的影响[J]. 口腔医学研究, 2019, 35(6): 541-545.
[10]	于阳, 徐文品, 焦琨, 刘冬蕊, 邢如霄, 叶佳朋, 张伟. 诱导大鼠脂肪来源干细胞向腺泡样细胞转化的实验研究[J]. 口腔医学研究, 2019, 35(5): 493-496.
[11]	汪晓龙,曹顺顺,张鹏,舒传继,夏林. 人源性长寿保障基因2在口腔鳞癌组织中表达及对细胞增殖和侵袭能力的影响[J]. 口腔医学研究, 2019, 35(2): 142-146.
[12]	陆蓓. 重组人骨形态发生蛋白-9对人牙周膜成纤维细胞成骨分化的影响[J]. 口腔医学研究, 2018, 34(8): 857-861.
[13]	周茜雯, 曾豪, 张嘉昕, 刘恭奇, 王飒, 汪烈. 箭根酮对LPS刺激牙周膜成纤维细胞增殖能力的影响[J]. 口腔医学研究, 2018, 34(6): 653-656.
[14]	张厚华, 范华俐. RNA干扰沉默钙蛋白酶1对口腔鳞状细胞癌细胞株HSC3增殖和迁移的影响[J]. 口腔医学研究, 2018, 34(2): 152-156.
[15]	裴浩, 夏冬景, 黄莹莹. STAT6在涎腺腺样囊性癌组织中的表达及对细胞增殖和侵袭的影响[J]. 口腔医学研究, 2018, 34(11): 1212-1216.