炎症与正常牙周膜来源牙周膜干细胞的成骨分化能力和自噬水平  

Osteogenic ability and autophagy level between normal and inflammatory periodontal ligament stem cells

作　　者：毛家奇 赵力如 杨冬茹[1] 胡永青 代博文 李淑娟[1] Mao Jiaqi;Zhao Liru;Yang Dongru;Hu Yongqing;Dai Bowen;Li Shujuan(First Department of Periodontology,Hebei Key Laboratory of Stomatology,Hebei Clinical Research Center for Oral Diseases,School and Hospital of Stomatology,Hebei Medical University,Shijiazhuang 050017,Hebei Province,China;Department of Orthodontics,Hebei Key Laboratory of Stomatology,Hebei Clinical Research Center for Oral Diseases,School and Hospital of Stomatology,Hebei Medical University,Shijiazhuang 050017,Hebei Province,China;Hebei Medical University,Shijiazhuang 050017,Hebei Province,China)

机构地区：[1]河北医科大学口腔医学院•口腔医院牙周一科,河北省口腔医学重点实验室,河北省口腔疾病临床医学研究中心,河北省石家庄市050017 [2]河北医科大学口腔医学院•口腔医院正畸科,河北省口腔医学重点实验室,河北省口腔疾病临床医学研究中心,河北省石家庄市050017 [3]河北医科大学,河北省石家庄市050017

出　　处：《中国组织工程研究》2025年第1期74-79,共6页Chinese Journal of Tissue Engineering Research

基　　金：河北省医学科学研究课题(20211078),项目负责人:赵力如;河北省医学科学研究课题(20230185),项目负责人:毛家奇;河北省卫健委2019年度政府资助省级临床医学优秀人才培养和基础课题研究项目(2019061441-2),项目负责人:李淑娟;2017年河北省财政厅老年病防治项目(361029),项目负责人:李淑娟。

摘　　要：背景:炎症影响牙周膜干细胞的成骨分化,同时牙周膜干细胞的成骨能力与自噬水平密切相关,但是炎症是否影响牙周膜干细胞成骨分化不同阶段的成骨能力与自噬水平尚未见相关报道。目的:探讨牙周膜干细胞在牙周炎和正常状态下的碱性磷酸酶表达和自噬水平。方法:分离培养健康人群与牙周炎患者的牙周膜干细胞,进行Vimentin、pan-CK和Stro-1荧光染色。正常和炎症牙周膜干细胞成骨分化3,7,14d时,Westernblot检测碱性磷酸酶、LC3B、Beclin1、ATG5的蛋白表达,real-time PCR检测碱性磷酸酶、骨唾液蛋白、骨钙素、Runx2、LC3B、Beclin1、ATG5的mRNA表达。结果与结论:(1)牙周膜干细胞内Stro-1阳性表达,Vimentin阳性表达,pan-CK阴性表达;(2)成骨分化3,7,14 d,与正常牙周膜干细胞相比,炎症牙周膜干细胞所形成的矿化结节明显减少(P<0.01),碱性磷酸酶的蛋白和mRNA表达明显降低(P<0.05),骨唾液蛋白、骨钙素、Runx2的mRNA表达明显降低(P<0.05);(3)成骨分化7,14 d,与正常牙周膜干细胞相比,炎症牙周膜干细胞的ATG5、LC3B与Beclin1蛋白和mRNA表达均明显降低(P<0.05)。结果表明,炎症可减少牙周膜干细胞的矿化结节形成和碱性磷酸酶的表达,削弱牙周膜干细胞成骨分化7,14 d的自噬潜能。BACKGROUND:Inflammation affects the osteogenic differentiation of periodontal ligament stem cells,and the osteogenic ability and autophagy level of periodontal ligament stem cells are closely related.However,there are no relevant reports on whether inflammation affects the osteogenic ability and autophagy level of periodontal ligament stem cells at different stages of osteogenic differentiation.OBJECTIVE:To explore alkaline phosphatase expression and autophagy periodontal ligament stem cells levels in periodontitis and normal conditions.METHODS:Periodontal ligament stem cells from normal and periodontitis patients were isolated and cultured,and underwent Vimentin,pan-CK,and Stro-1 fluorescence staining.At 3,7,and 14 days of osteogenic differentiation,western blot assay was used to detect the protein expression levels of alkaline phosphatase,LC3B,Beclin1,and ATG5 in normal and inflammatory periodontal ligament stem cells.The mRNA expression levels of alkaline phosphatase,bone sialoprotein,osteocalcin,Runx2,LC3B,Beclin1,and ATG5 were detected by real-time PCR.RESULTS AND CONCLUSION:(1)Stro-1 was positive,Vimentin was positive,and pan CK was negative in periodontal ligament stem cells.(2)At 3,7,and 14days after osteogenic differentiation,compared with normal periodontal ligament stem cells,the mineralization nodules formed by periodontal ligament stem cells from inflammatory sources were significantly reduced(P<0.01);the expression of alkaline phosphatase protein and mRNA was significantly lower(P<0.05);the mRNA expression levels of bone sialoprotein,osteocalcin,and Runx2 were significantly decreased(P<0.05).(3)At 7 and 14 days after osteogenic differentiation,compared with normal periodontal ligament stem cells,the expression levels of ATG5,LC3B,and Beclin1 proteins and mRNA of periodontal ligament stem cells were downregulated(P<0.05).These findings suggest that inflammation reduces the activity of periodontal ligament stem cells in mineralizing nodule formation and the expression of alkaline phosphatase and weake

关 键 词：牙周膜干细胞 牙周炎 碱性磷酸酶 自噬 炎症 成骨分化 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R781.4