口腔医学研究

口腔医学研究 ›› 2022, Vol. 38 ›› Issue (1): 65-70.DOI: 10.13701/j.cnki.kqyxyj.2022.01.014

• 牙周病学研究 • 上一篇    下一篇

牙龈卟啉单胞菌对破骨前体细胞分化的调控及其机制研究

吴姝萱1, 谭晶晶1, 李锋1, 万启龙1,2*   

  1. 1.武汉大学口腔医学院口腔基础医学省部共建国家重点实验室培育基地和口腔生物医学教育部重点实验室 湖北 武汉 430079;
    2.武汉大学口腔医院正颌与唇腭裂整形外科 湖北 武汉 430079
  • 收稿日期:2021-09-09 出版日期:2022-01-28 发布日期:2022-01-21
  • 通讯作者: * 万启龙,E-mail:qilong.wan@whu.edu.cn
  • 作者简介:吴姝萱(1997~ ),女,河南商丘人,硕士,主要从事口腔颌面外科的临床和基础研究。
  • 基金资助:
    湖北省自然科学基金面上项目(编号:2020CFB841)

Role and Mechanism of Porphyromonas Gingivalis in the Differentiation of Osteoclast Precursor

WU Shuxuan1, TAN Jingjing1, LI Feng1, WAN Qilong1,2*   

  1. 1. The State Key Laboratory Breeding Base of Basic Science of Stomatology & Key Laboratory of Oral Biomedicine Ministry of Education, School of Stomatology, Wuhan University, Wuhan 430079, China;
    2. Department of Orthognathic & Cleft Lip and Palate Plastic Surgery, Hospital of Stomatology, Wuhan University, Wuhan 430079, China
  • Received:2021-09-09 Online:2022-01-28 Published:2022-01-21

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摘要： 目的:探究牙龈卟啉单胞菌(Porphyromonas gingivalis, P.g)在体外直接共培养刺激和在体内远处局部刺激对破骨前体细胞(osteoclast precursors,OCPs)分化的调控及其机制。方法:提取培养6~8周雄性C57/BL6小鼠股骨OCPs。将OCPs与不同感染复数的P.g直接共培养6 h后,检测并比较各组OCPs的活性及亚型比例。去除P.g后,继续诱导OCPs向破骨细胞分化。于小鼠右侧股骨制造骨缺损,并在骨缺损处注射含或不含P.g的悬液,每周两次。4周后提取培养小鼠左侧股骨OCPs,并诱导OCPs向破骨细胞分化。分别在各组OCPs培养的第4天和第6天,检测并比较各组间的成熟破骨细胞数量、相关基因和蛋白的表达水平。结果:P.g的体外直接共培养刺激不影响OCPs的活性及各亚型比例,但抑制了OCPs向破骨细胞分化,这与P.g刺激后OCPs的破骨分化相关基因和蛋白均下降有关。P.g的体内远处局部刺激,上调了OCPs部分与破骨分化和炎性相关基因的表达,但OCPs向破骨细胞分化的能力不受其影响。结论:P.g的体外直接共培养刺激会抑制OCPs向破骨细胞分化,而体内局部刺激不影响其他部位OCPs向破骨细胞分化的能力。

关键词: 牙龈卟啉单胞菌, 破骨前体细胞, 破骨细胞, 直接共培养, 体内局部刺激

Abstract: Objective: To investigate the role and mechanism of Porphyromonas gingivalis(P.g)in the differentiation of osteoclast precursors (OCPs) through in vitro direct co-culture or in vivo remote local stimulation. Methods: The OCPs were isolated from 6 to 8-week-old male C57BL/6 mice femurs and cultured in vitro. The OCPs were direct co-cultured with different multiplicity of infection P.g for 6 hours. Then the viability and the ratio of subtypes of OCPs in each group were detected and compared. After removing P.g, the OCPs continued to be cultured to induce osteoclastogenesis. Bone defects were created on the right femurs of the mice, and the suspension with or without P.g was injected into the defects twice a week for 4 weeks. The OCPs from left femurs of mice were isolated, cultured, and induced osteoclastogenesis. After culturing OCPs for 4 or 6 days, the number of mature osteoclasts and the expression levels of related genes and proteins in each group were detected and compared. Results: Direct in vitro co-culture of P.g and OCPs did not affect the activity and the proportion of subtypes of OCPs, but inhibited the osteoclastogenesis of OCPs which was related to the declining expression of osteoclast-related genes and proteins. The remote in vivo local stimulation of P.g up-regulated the expression levels of some osteoclast-related and inflammation-related genes, but did not affect the osteoclastogenesis of OCPs. Conclusion: Direct in vitro co-culture of P.g and OCPs can inhibit the osteoclastogenesis of OCPs, while in vivo local stimulation does not affect the osteoclastogenesis of OCPs in other parts of the body.

Key words: Porphyromonas gingivalis, osteoclast precursors, osteoclast, direct co-culture, in vivo local stimulation