M2型巨噬细胞来源外泌体通过抑制NLRP3炎症小体活化介导的细胞焦亡促进人牙周膜干细胞成骨分化
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达州市中西医结合医院

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四川省卫健委卫生健康科研课题(23PJ421)


M2-type Macrophage-derived Exosomes Promote Osteogenic Differentiation of Human Periodontal Ligament Stem Cells by Inhibiting Pyroptosis Mediated by NLRP3 Inflammasome Activation
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    目的 探究M2型巨噬细胞来源外泌体(M2-type macrophage-derived exosomes, M2-Exo)对脂多糖(lipopolysaccharide, LPS)诱导的人牙周膜干细胞(human periodontal ligament stem cells, hPDLSCs)成骨分化的作用及其机制研究。方法 以RAW264.7和hPDLSCs细胞为研究对象。诱导RAW264.7细胞向M2型巨噬细胞极化并提取M2-Exo,采用流式细胞术、Western blot(WB)、透射电镜等方法进行鉴定。以hPDLSCs细胞为研究对象,首先采用LPS诱导构建牙周炎模型,随后将M2-Exo与hPDLSCs细胞进行共培养(共培养期间更换含新鲜M2-Exo的成骨诱导培养基),通过碱性磷酸酶(alkaline phosphatase,ALP)染色、茜素红S法(alizarin red?S staining,ARS)染色及WB等方法进行分析,探究M2-Exo的调控作用。加入Nigericin(NLRP3激动剂)就行回复实验。结果 成功获得M2型巨噬细胞及M2-Exo;M2-Exo可显著逆转LPS对hPDLSCs细胞成骨分化的抑制,提升ALP活性、促进矿化结节形成,上调成骨相关标志物表达(P < 0.001);同时缓解LPS诱导的细胞损伤,降低促炎因子水平,抑制NOD样受体家族含吡啶结构域蛋白3(NODlike receptor family pyrin domain containing 3,NLRP3)介导的细胞焦亡(P < 0.001);Nigericin可逆转M2-Exo的抑焦亡及促成骨作用(P < 0.05)。结论 M2-Exo通过抑制NLRP3炎症小体活化介导的细 胞焦亡,缓解LPS诱导的hPDLSCs细胞损伤,进而促进其成骨分化,为牙周炎临床治疗及牙周组织再生提供了新型干预策略。

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  • 收稿日期:2026-05-14
  • 最后修改日期:2026-06-25
  • 录用日期:2026-07-02
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