硫化氢促进炎症微环境中正畸力作用下的牙周组织成骨分化

口腔生物医学 ›› 2022, Vol. 13 ›› Issue (1): 20-25.

硫化氢促进炎症微环境中正畸力作用下的牙周组织成骨分化

赵逸,王月,华咏梅,廖崇珊

同济大学附属口腔医院

收稿日期:2021-11-15 修回日期:2022-01-23 出版日期:2022-03-25 发布日期:2022-03-21
通讯作者: 廖崇珊 E-mail:liaochongshan@163.com
基金资助:
国家自然科学基金;中华口腔医学会青年临床科研基金口腔正畸研究项目

Hydrogen sulfide improves orthodontic force-induced osteogenesis of periodontal tissue in inflammatory microenvironment

Received:2021-11-15 Revised:2022-01-23 Online:2022-03-25 Published:2022-03-21
Contact: Chongshan LIAO E-mail:liaochongshan@163.com
Supported by:
National Natural Science Foundation of China

摘要/Abstract

摘要： 目的：研究外源性硫化氢（H2S）对牙周炎大鼠正畸牙移动中骨形成和炎症微环境中人牙周膜细胞（hPDLCs）成骨分化的影响。方法：将48只SD大鼠随机分为对照组、正畸治疗（OT）组、牙周炎正畸治疗（P+OT）组、牙周炎正畸+硫氢化钠（NaHS，H2S供体）（P+OT+NaHS）组，通过显微CT扫描、骨钙素（OCN）免疫组化染色和碱性磷酸酶（ALP）染色观察牙周炎状态下正畸牙张力侧的骨形成，通过实时定量RT-PCR检测牙周炎状态下正畸张力侧牙周组织中成骨相关因子的表达。体外实验分为对照组、张力（T）组、脂多糖（LPS）+T组、LPS+T+NaHS组，通过ALP和茜素红染色观察LPS诱导的hPDLCs在张力下的成骨分化，通过实时定量RT-PCR检测炎症环境中张力作用下hPDLCs成骨相关因子的表达。结果：体内实验发现P+OT+NaHS组张力侧骨组织参数高于P+OT组（P<0.05）；与P+OT组相比，P+OT+NaHS组中Col1、Alp和Ocn成骨相关基因及蛋白表达也显著增加（P<0.05）。体外实验表明，与LPS+T组相比，LPS+T+NaHS组的ALP和茜素红染色较深，OD值较高（P<0.05），COL1、ALP、OCN表达均显著增加（P<0.05）。结论：NaHS可维持牙周组织中的骨矿物质密度，且上调受炎症抑制的成骨因子的mRNA表达，促进炎症微环境下正畸力诱导的骨形成。H2S可能在促进牙周炎环境下正畸牙的骨形成方面具有重要意义。

Abstract: Objective:This study investigated the effects of H2S on orthodontic bone formation in rats with periodontitis and osteogenic differentiation of human periodontal ligament cells (hPDLCs) in the inflammatory microenvironment. Methods:Forty-eight rats were divided into four groups: phosphate-buffered saline control group, orthodontic treatment (OT) group, ligature induced-periodontitis+OT group, ligature induced-periodontitis+OT with NaHS (H2S donor) group. Bone formation at tension sites of periodontal tissue was observed by micro-computed tomography (micro-CT), immunohistochemistry staining for osteocalcin (OCN) and alkaline phosphatase (ALP) staining. Expression of osteogenesis related factors were examined by real-time quantitative PCR. In vitro, experiments were divided into control group, tension (T) group, lipopolysaccharide+T group, lipopolysaccharide+T with NaHS group. Real-time quantitative PCR was used to detect the expression of hPDLCs osteogenesis-related factors under tension force in the inflammatory environment. Results:It was found that the bone tissue parameters of the tension side in the P+OT+NaHS group were higher than those in the P+OT group in vivo. Compared with the P+OT group, the expression of Col1, Alp and Ocn osteogenesis-related genes and proteins were significantly increased in the P+OT+NaHS group（P<0.05）. At the same time, it was showed that the LPS+T+NaHS group, compared with the LPS+T group, had deeper staining results, higher OD value and significantly increased expressions of COL1, ALP and OCN osteogenesis-related genes in vitro（P<0.05）. Conclusions:NaHS could maintain bone mineral density in periodontal tissue, up-regulate the mRNA expression of osteogenic factors inhibited by inflammation, and promote orthodontic force-induced bone formation in an inflammatory microenvironment. Our findings suggested that H2S may play an important role in promoting bone formation of orthodontic teeth in periodontitis environment.

中图分类号:

R781.4

赵逸王月华咏梅廖崇珊. 硫化氢促进炎症微环境中正畸力作用下的牙周组织成骨分化[J]. 口腔生物医学, 2022, 13(1): 20-25.

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