中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (7): 1051-1056.doi: 10.12307/2022.143

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

柚皮素干预人牙周膜干细胞的成骨分化能力

罗小玲1，张  丽1，杨茂桦1，徐  洁2，徐晓梅2   

  1. 1西南医科大学口颌面修复重建和再生实验室，四川省泸州市  646000；2西南医科大学附属口腔医院正畸科，四川省泸州市  646000
  • 收稿日期:2020-10-22 修回日期:2020-10-24 接受日期:2020-11-21 出版日期:2022-03-08 发布日期:2021-10-29
  • 通讯作者: 徐晓梅，博士，主任医师，副教授，西南医科大学附属口腔医院正畸科，四川省泸州市 646000
  • 作者简介:罗小玲，女，1994年生，四川省资中县人，汉族，西南医科大学在读硕士，主要从事口腔正畸学的研究。
  • 基金资助:
    四川省大学生创新创业训练计划项目(S202010632063)，项目负责人：杨茂桦；泸州市市-校联合项目(2020LZXNYDZ06)，项目负责人：徐晓梅

Effect of naringenin on osteogenic differentiation of human periodontal ligament stem cells

Luo Xiaoling1, Zhang Li1, Yang Maohua1, Xu Jie2, Xu Xiaomei2   

  1. 1Oral & Maxillofacial Reconstruction and Regeneration Laboratory, Southwest Medical University, Luzhou 646000, Sichuan Province, China; 2Department of Orthodontics, Affiliated Stomatology Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • Received:2020-10-22 Revised:2020-10-24 Accepted:2020-11-21 Online:2022-03-08 Published:2021-10-29
  • Contact: Xu Xiaomei, MD, Chief physician, Associate professor, Department of Orthodontics, Affiliated Stomatology Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • About author:Luo Xiaoling, Master candidate, Oral & Maxillofacial Reconstruction and Regeneration Laboratory, Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • Supported by:
    the Innovation and Entrepreneurship Training Program for College Students in Sichuan Province, No. S202010632063 (to YMH); the Luzhou City-School Joint Project, No. 2020LZXNYDZ06 (to XXM)

PDF

58

摘要：

文题释义：
柚皮素：是广泛存在于芸香科植物中的一种二氢黄酮类化合物，能促进成骨细胞的增殖和分化，抑制破骨细胞的活性和生成。
牙周膜干细胞：是存在于牙周膜中的一种未分化的间充质细胞，具有高增殖能力和多能性，能定向分化为成骨细胞，其成骨分化能力受细胞因子、激素、药物等诸多因素的调节。
背景：柚皮素具有抗菌、抗炎、抗纤维化、抗氧化等多种生理活性，被广泛应用于医药和食品领域。近来有研究证明，柚皮素能有效促进间充质干细胞成骨分化，但柚皮素是否有调节牙周膜干细胞的成骨分化能力尚不清楚。
目的：探究不同浓度柚皮素对人牙周膜干细胞成骨分化能力的影响。
方法：原代分离培养人牙周膜干细胞，分别用含浓度为10，100 nmol/L、1，10，100 μmol/L和1 mmol/L柚皮素的α-MEM培养基处理72 h后，CCK-8法检测柚皮素对人牙周膜干细胞的细胞毒性。取第3代人牙周膜干细胞，分别用含0，1，10，100 μmol/L柚皮素的成骨诱导液培养，诱导3，5，7 d后分别进行碱性磷酸酶染色和碱性磷酸酶活性检测，诱导7 d后Real-time PCR检测成骨相关因子Runx2、骨桥蛋白和骨钙素的表达，诱导14 d后进行茜素红染色和矿化结节的定量分析。
结果与结论：10 nmol/L-100 μmol/L柚皮素对人牙周膜干细胞没有细胞毒性，1 mmol/L柚皮素对人牙周膜干细胞有明显的细胞毒性；1-100 μmol/L柚皮素显著促进碱性磷酸酶活性和矿物质沉积，上调Runx2、骨桥蛋白和骨钙素的基因表达。结果表明，柚皮素能显著促进人牙周膜干细胞的成骨分化，100 μmol/L时促成骨分化能力最强。

https://orcid.org/0000-0003-1935-5824(罗小玲)；https://orcid.org/0000-0003-2211-655X(徐晓梅) 

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: 干细胞, 人牙周膜干细胞, 柚皮素, 细胞毒性, 成骨, 分化, 因子

Abstract: BACKGROUND: Naringenin has various physiological activities such as anti-bacterial, anti-inflammatory, anti-fibrosis, and anti-oxidation, and is widely used in the fields of medicine and food. Recent studies have shown that naringenin can effectively promote the osteogenic differentiation of mesenchymal stem cells, but it is unclear whether naringenin regulates the osteogenic differentiation of periodontal ligament stem cells.
OBJECTIVE: To investigate the effects of naringenin with different concentrations on the osteogenic differentiation of human periodontal ligament stem cells. 
METHODS: The primary human periodontal ligament stem cells were isolated and cultured. After treatment with naringenin at concentrations of 10, 100 nmol/L, 1, 10, 100 μmol/L and 1 mmol/L for 72 hours, the cytotoxicity of naringenin on periodontal ligament stem cells was measured via CCK-8 assays. The third generation of human periodontal ligament stem cells was cultured with osteogenic medium containing 0, 1, 10 and 100 μmol/L naringenin respectively. Alkaline phosphatase staining and activity detection were performed after 3, 5, and 7 days. Real-time PCR was used to detect the expression of osteogenic factors including Runx2, osteopontin and osteocalcin in human periodontal ligament stem cells after induction for 7 days. Alizarin red staining and quantitative determination of mineralized nodule were performed after induction for 14 days. 
RESULTS AND CONCLUSION: Naringenin at a concentration of 10 nmol/L-100 μmol/L has no cytotoxicity to human periodontal ligament stem cells, and naringenin at a concentration of 1 mmol/L has obvious cytandotoxicity to human periodontal ligament stem cells. An appropriate concentration of naringenin (1-100 μmol/L) significantly promotes the increase of alkaline phosphatase activity and mineral deposition and upregulates the gene expression of Runx2, osteopontin and osteocalcin. The results showed that naringenin can significantly promote the osteogenic differentiation of human periodontal ligament stem cells, and the ability to promote osteogenic differentiation is strongest at 100 μmol/L. 


Key words: stem cells, human periodontal ligament stem cells, naringenin, cytotoxicity, osteoggenesis, differentiation, factor

中图分类号: