成骨诱导人牙周膜干细胞来源外泌体促进炎症微环境下人牙周膜干细胞成骨分化  

Osteogenesis-induced exosomes derived from human periodontal ligament stem cells promote osteogenic differentiation of human periodontal ligament stem cells in an inflammatory microenvironment

作　　者：艾克帕尔·艾尔肯 陈晓涛 吾凡别克·巴合提[2] Aikepaer·Aierken;Chen Xiaotao;Wufanbieke·Baheti(School of Life Sciences,Xinjiang University,Urumqi 830000,Xinjiang Uygur Autonomous Region,China;Department of Stomatology,People’s Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830000,Xinjiang Uygur Autonomous Region,China)

机构地区：[1]新疆大学生命科学院,新疆维吾尔自治区乌鲁木齐市830000 [2]新疆维吾尔自治区人民医院口腔科,新疆维吾尔自治区乌鲁木齐市830000

出　　处：《中国组织工程研究》2025年第7期1388-1394,共7页Chinese Journal of Tissue Engineering Research

摘　　要：背景:成骨诱导间充质干细胞来源外泌体具有较强的成骨分化能力,但是在炎症微环境下对人牙周膜干细胞成骨分化的影响尚不明确。目的:探究成骨诱导人牙周膜干细胞来源外泌体在炎症微环境下对人牙周膜干细胞成骨分化的影响。方法:收集离体牙并分离培养人牙周膜干细胞,成骨诱导3 d后提取外泌体。将人牙周膜干细胞分为4组:对照组加入成骨诱导培养基,外泌体组加入含5μg/mL外泌体的成骨诱导培养基,炎症模型和炎症模型+外泌体组以1μg/mL脂多糖处理24 h构建细胞炎症微环境,炎症模型组在脂多糖处理后加入成骨诱导培养基,炎症模型+外泌体组在脂多糖处理后加入含5μg/mL外泌体的成骨诱导培养基。通过茜素红以及碱性磷酸酶染色法检测各组人牙周膜干细胞的成骨分化能力;实时荧光定量PCR与免疫印迹法检测各组人牙周膜干细胞中Runt相关转录因子2、骨桥蛋白、成骨细胞特异性转录因子Osterix(OSX)和wnt通路相关蛋白β-catenin的表达。结果与结论:(1)与对照组相比,炎症模型组碱性磷酸酶染色相对面积、矿化结节染色相对面积以及Runt相关转录因子2、骨桥蛋白、OSX的表达量显著降低(P <0.05);(2)与炎症模型组相比,炎症模型+外泌体组碱性磷酸酶染色相对面积、矿化结节染色相对面积以及Runt相关转录因子2、骨桥蛋白、OSX的表达显著升高(P <0.05);(3)与对照组相比,炎症模型组wnt通路相关蛋白β-catenin表达量显著增加(P <0.05);与炎症模型组相比,炎症模型+外泌体组β-catenin表达量显著降低(P <0.05)。结果表明,成骨诱导人牙周膜干细胞来源外泌体可促进炎症微环境下人牙周膜干细胞的成骨分化,其作用机制可能与wnt/β-catenin信号通路有关。BACKGROUND:The osteogenic differentiation ability of exosomes derived from osteogenic mesenchymal stem cells is well established.However,their impact on the osteogenic differentiation of human periodontal ligament stem cells in an inflammatory microenvironment remains unclear.OBJECTIVE:To examine the impact of exosomes derived from osteogenesis-induced human periodontal ligament stem cells on the osteogenic differentiation of human periodontal ligament stem cells within an inflammatory microenvironment.METHODS:Human periodontal ligament stem cells were isolated and cultured.After 3 days of osteogenic induction,exosomes were extracted.Human periodontal ligament stem cells were divided into four groups.Control group was treated with osteogenesis-induced medium.The exosome group was treated with osteogenesis-induced medium containing 5 μg/mL exosomes.Inflammatory model and inflammatory model + exosome groups were treated with 1 μg/mL lipopolysaccharide for 24 hours to construct a cellular inflammatory microenvironment.The inflammatory model group was treated with osteogenesisinduced medium after lipopolysaccharide intervention.The inflammatory model + exosome group was treated with osteogenesis-induced medium containing 5 μg/mL exosome.The osteogenic differentiation ability of human periodontal ligament stem cells was assessed using alkaline phosphatase staining and alizarin red staining.The expressions of Runt-related transcription factor 2,osteopontin,osteoblast-specific transcription factor Osterix(OSX) and wnt pathway-related protein β-catenin were detected by real-time fluorescence quantitative PCR and western blot assay.RESULTS AND CONCLUSION:(1) Compared with the control group,the relative area stained by alkaline phosphatase,the relative area stained by mineralized nodules and the expression levels of Runx2,osteopontin,and OSX were significantly decreased in the inflammatory model group(P < 0.05).(2) Compared with the inflammatory model group,the expression of Runx2,osteopontin,and OSX in the inflammator

关 键 词：人牙周膜干细胞 外泌体 炎症微环境 成骨分化 成骨细胞 信号通路 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R34