口腔医学研究 ›› 2024, Vol. 40 ›› Issue (3): 214-220.DOI: 10.13701/j.cnki.kqyxyj.2024.03.005

• 口腔生物学研究 • 上一篇    下一篇

芒柄花素促进牙囊干细胞成骨向分化

陶天翼1, 李正强2, 郑晓雪3, 韩冰1*   

  1. 1.吉林大学口腔医院口腔颌面外科 吉林省牙发育及颌骨重塑与再生重点实验室 吉林 长春 130021;
    2.南方医科大学口腔医院口腔颌面外科 广东 广州 510515;
    3.吉林省肿瘤医院耳鼻咽喉口腔颌面头颈外科 吉林 长春 130012
  • 收稿日期:2023-11-06 出版日期:2024-03-28 发布日期:2024-03-25
  • 通讯作者: * 韩冰,E-mail:hbing@jlu.edu.cn
  • 作者简介:陶天翼(1996~ ),男,吉林人,硕士在读,医师,研究方向:口腔颌面部骨缺损的修复重建及再生。
  • 基金资助:
    吉林省科技发展计划项目(编号:20230204079YY)

Formononetin Motivates Osteogenic Differentiation of Dental Follicle Stem Cells

TAO Tianyi1, LI Zhengqiang2, ZHENG Xiaoxue3, HANG Bing1*   

  1. 1. Department of Oral and Maxillofacial Surgery, Jilin University Stomatological Hospital, Key Laboratory of Tooth Development and Craniofacial Reconstruction and Regeneration, Changchun 130021, China;
    2. Department of Oral and Maxillofacial Surgery, Stomatology Hospital, Southern Medical University, Guangzhou 510515, China;
    3. Department of Otolaryngology, Oral-Maxillofacial Head and Neck Surgery, Jilin Province Cancer Hospital, Changchun 130012, China
  • Received:2023-11-06 Online:2024-03-28 Published:2024-03-25

摘要: 目的:研究芒柄花素对牙囊干细胞(human dental follicle stem cells,hDFSCs)成骨分化的影响。方法:首先对hDFSCs进行分离、培养及鉴定,并使用含不同浓度芒柄花素(0、0.1、1、10 μmol/L)培养基培养细胞。通过CCK-8实验、结晶紫染色和活/死细胞荧光染色评估细胞的活性状况;通过细胞骨架荧光染色观察细胞骨架状况;采用RT-qPCR检测细胞成骨相关基因的表达变化;利用碱性磷酸酶(alkaline phosphatase,ALP)染色及活性检测法评估细胞成骨分化过程中ALP活性的影响;通过茜素红染色法评估细胞成骨分化过程中钙化结节数量的影响。结果:hDFSCs具有干细胞特性;不同浓度的芒柄花素对hDFSCs活性无显著性影响;1 μmol/L的芒柄花素可显著促进Runx2、OCN、Col-Iα1的mRNA表达;并能够提高的细胞内ALP活性和钙化结节的数量。结论:1 μmol/L芒柄花素可促进hDFSCs的成骨分化。

关键词: 芒柄花素, 牙囊干细胞, 成骨分化

Abstract: Objective: To elucidate the effect of formononetin on osteogenic differentiation of human dental follicle stem cells (hDFSCs). Methods: hDFSCs were isolated, cultured, and identified. The cells were treated with different concentrations of formononetin (0, 0.1, 1, and 10 μmol/L) in the culture medium. Cell viability was assessed by CCK-8 assay, crystal violet staining, and live/dead cell fluorescence staining. The cell cytoskeleton was examined using fluorescent staining. The expression of osteogenic-related genes was analyzed through RT-qPCR. Alkaline phosphatase (ALP) activity was assessed to evaluate the osteogenic differentiation of cells. The influence of formononetin on calcium nodule formation during osteogenic differentiation was evaluated using alizarin red staining. Results: hDFSCs exhibited stem cell characteristics. Different concentrations of formononetin had no significant effect on hDFSCs viability. Whereas, 1 μmol/L formononetin facillitated the expression of Runx2, OCN, and Col-Iα1 mRNA. Additionally, 1 μmol/L formononetin enhanced the intracellular ALP activity and the number of calcium nodules. Conclusion: Formononetin at 1 μmol/L promotes the osteogenic differentiation of hDFSCs.

Key words: formononetin, human dental follicle stem cells, osteogenesis differentiation