摘要
目的:探讨M2巨噬细胞外泌体(exosomes derived from M2 macrophages,M2-Exos)对牙髓干细胞(dental pulp stem cells,DPSCs)和人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)成血管能力的影响,以评估其在牙髓再生中的促血管效应及应用潜力。方法:差速离心法分别提取M1巨噬细胞外泌体(exosomes drived from M1 macrophages,M1-Exos)与M2-Exos,经透射电子显微镜(transmission electron microscope,TEM)、纳米颗粒跟踪分析(nanoparticle tracking analysis,NTA)和蛋白印迹(Western blotting,WB)鉴定。PKH-26标记两种外泌体后与DPSCs、HUVECs共培养,观察摄取情况。通过Transwell迁移实验检测细胞迁移能力。采用基质胶小管形成实验评估成血管能力。结果:本研究成功分离鉴定符合标准的M1-Exos与M2-Exos,两者均可被DPSCs和HUVECs有效摄取。与M1-Exos抑制迁移不同,M2-Exos显著促进DPSCs和HUVECs迁移(P<0.05)。M2-Exos促进DPSCs内皮细胞分化,诱导血管样管腔结构形成,显著增强HUVECs成血管潜能,形成规则有序的毛细血管样结构(P<0.05)。结论:M2-Exos可在体外明显增强DPSCs和HUVECs的迁移能力,诱导DPSCs内皮细胞分化,显著促进体外HUVECs血管发生,提示M2-Exos具有优异的成血管潜能。
Objective:This study was designed to investigate the effects of M2 macrophage-derived exosomes(M2-Exos)on the angiogenic potential of dental pulp stem cells(DPSCs)and human umbilical vein endothelial cells(HUVECs),and to evaluate their proangiogenic competency and application potential in dental pulp regeneration.Methods:M1-Exos and M2-Exos were extracted by differential centrifugation and identified by transmission electron microscope(TEM),nanoparticle tracking analysis(NTA),and western blotting(WB).Next,PKH-26-labeled exosomes were co-cultured with DPSCs and HUVECs to observe uptake efficiency.Subsequently,the effect of M1-Exos and M2-Exos on the migration capability of DPSCs and HUVECs was assessed by transwell migration assay.Finally,angiogenic capacity was evaluated using Matrigel tube formation assay.Results:In this study,M1-Exos and M2-Exos meeting exosomal standards were successfully isolated and identified.Meanwhile,the two exosomes could be effectively taken up by DPSCs and HUVECs.In addition,compared to M1-Exos,which significantly inhibited the migration of DPSCs and HUVECs,the number of migrated cells was considerably increased after M2-Exos application(P<0.05).Importantly,in sharp contrast with M1-Exos,M2-Exos not only promoted the endothelial cell differentiation of DPSCs and induced the formation of vascular-like tubular structures,but also remarkably enhanced the angiogenic potential of HUVECs to generate regular and orderly capillary-like structures(P<0.05).Conclusion:M2-Exos could significantly facilitate the migration of DPSCs and HUVECs in vitro.Furthermore,they induced the differentiation of endothelial cells of DPSCs and promoted the angiogenesis of HUVECs,suggesting that M2-Exos possess desirable proangiogenic property.
作者
王宇洁
汪一帆
段大洁
刘甄苑
胡筱涵
毛靖
石鑫
WANG Yu-jie;WANG Yi-fan;DUAN Da-jie;LIU Zhen-yuan;HU Xiao-han;MAO Jing;SHI Xin(Center of Stomatology,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Hubei Wuhan 430030,China;School of Stomatology,Tongji Medical College,Huazhong University of Science and Technology,Hubei Wuhan 430030,China;Hubei Province Key Laboratory of Oral and Maxillofacial Development and Regeneration,Hubei Wuhan 430022,China;Outpatient Department Office,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Hubei Wuhan 430030,China)
出处
《临床口腔医学杂志》
2025年第9期515-520,共6页
Journal of Clinical Stomatology
基金
国家自然科学基金项目(82401074)
湖北省教学改革研究项目(HBJG-250012)
湖北省大学生创新基金项目(S202510487620)
华中科技大学研究生创新基金项目(YCJJ20252415)
华中科技大学大学生创新基金项目(X202510487134)
华中科技大学同济医学院第二临床学院教学研究基金项目(2022058)
华中科技大学同济医学院附属同济医院科研基金项目(24-2KYC13061-02)。
关键词
再生性牙髓治疗
血运重建
巨噬细胞
外泌体
Regenerative endodontic therapy
Revascularization
Macrophage
Exosome
