人脐带间充质干细胞来源小胞外囊泡用于治疗放创复合型骨损伤的疗效研究OA
Therapeutic efficacy of human umbilical cord mesenchymal stem cell-derived small extracellular vesicles against radiation-combined bone injury
目的 构建放创复合型骨损伤(RCBI)大鼠模型,评估人脐带间充质干细胞来源小胞外囊泡(hUC-MSC-sEV)的治疗效应,并探讨其促进骨修复作用.方法 采用超速离心法分离 hUC-MSC-sEV,通过纳米颗粒追踪分析(NTA)、透射电镜(TEM)及Western印迹进行鉴定.将SD大鼠随机分为3组:未经电离辐射(IR)对照组(non-IR)、电离辐射模型组(IR)及IR后接受sEV治疗组(IR+sEV).利用⁶⁰Co γ射线(9.0 Gy)局部辐射联合胫骨钻孔手术建立RCBI模型.IR+sEV组分别于造模后第1天和第8天接受sEV局部皮下注射.采用微型CT(MicroCT)动态监测术后第7、14、21天的骨修复进程,定量分析骨骼矿物质密度(BMD)、骨体积(BV)及骨表面积体积比(BS/BV);术后第21天取材行苏木素-伊红(HE)染色和马松三色(Masson)染色,评估组织形态学改变.结果 分离所得sEV的平均粒径约163.4 nm,呈典型杯状结构,特异性表达肿瘤易感基因101蛋白(TSG101)和CD81标志蛋白.IR组骨修复显著延迟,第21天仍见较大缺损;与IR组比较,IR+sEV组骨修复进程显著改善,第14天已可见明显的新生骨形成,第21天骨缺损基本愈合.定量分析显示,IR+sEV组的新生骨BMD高于IR组、新生BV大于IR组,BS/BV低于IR组(P<0.05).组织学观察结果表明,IR+sEV组新生骨小梁结构更为完整,胶原纤维排列趋于规则有序.结论 hUC-MSC-sEV充质干细胞sEV显著促进RCBI修复,为放射性骨损伤的生物学治疗提供了新依据.
Objective To establish a rat model of radiation-combined bone injury(RCBI),evaluate the therapeutic efficacy of human umbilical cord mesenchymal stem cell-derived small extracellular vesicles(hUC-MSC-sEVs),and to investigate their role in promoting bone repair.Methods hUC-MSC-sEVs were isolated via ultracentrifugation and characterized using nanoparticle tracking analysis(NTA),transmission electron microscopy(TEM),and Western blotting.SD rats were randomly divided into three groups:non-ionizing radiation(IR)control group(non-IR),IR model group(IR),and sEVs-treated after IR group(IR+sEV).The RCBI model was established via local irradiation with⁶⁰Co γ-rays(9.0 Gy)combined with tibial drilling injury.The IR+sEV group received local sEVs injections on days 1 and 8 post-irradiation.Bone repair was dynamically monitored by micro computed tomography(MicroCT)on days 7,14 and 21 while bone mineral density(BMD),bone volume(BV),and bone surface/volume ratio(BS/BV)were quantitatively analyzed.On day 21,tibial specimens were harvested for hematoxylin-eosin staining and Masson's Trichrome staining to assess the histomorphological changes.Results The isolated sEVs had a mean particle size of approximately 163.4 nm,were typically cup-shaped morphologically,and expressed specific marker proteins-tumor susceptibility gene 101 protein(TSG101)and cluster of differentiation 81(CD81).Bone repair was significantly delayed in the IR group,with large defects still visible on day 21,but was markedly enhanced in the IR+sEV group,as evidenced by new bone formation observed as early as day 14 and nearly complete defect healing by day 21.Quantitative analysis found that BMD,BV and BS/BV were all significantly superior in the IR+sEV group to those of the IR group(P<0.05).Histological examination revealed that trabecular bone structures were more intact and collagen fibers were more orderly in arrangement in the IR+sEV group.Conclusion Human umbilical cord mesenchymal stem cell-derived sEVs can significantly boost the repair of radiation-combined bone injury,which is expected to provide new insight for the biological treatment of radiation-induced bone damage.
魏正阳;李雪;张昭玮;郑远静;王洋;刘元林;张毅
军事科学院军事医学研究院,北京 100850军事科学院军事医学研究院,北京 100850军事科学院军事医学研究院,北京 100850军事科学院军事医学研究院,北京 100850军事科学院军事医学研究院,北京 100850军事科学院军事医学研究院,北京 100850军事科学院军事医学研究院,北京 100850
医药卫生
放创复合型骨损伤间充质干细胞小胞外囊泡骨重塑无细胞治疗
radiation-combined bone injurymesenchymal stem cellsmall extracellular vesiclebone remodelingcell-free therapy
《军事医学》 2026 (5)
351-357,7
国家科技资源共享服务平台细胞疗法研究计划(PT-2024-04)
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