脂肪间充质干细胞来源外泌体对施万细胞增殖功能及早期神经再生特征影响的研究  

Effects of adipose-derived mesenchymal stem cell exosomes on Schwann cell proliferation and early nerve regeneration

作　　者：任致奇 尚爱加 杨博尧[1,3] 柏钧 贾志博 刘修志 王玉 REN Zhiqi;SHANG Aijia;YANG Boyao;BO Jun;JIA Zhibo;LIU Xiuzhi;WANG Yu(Chinese PLA Medical School,Beijing 100853,China;Department of Neurosurgery,the First Medical Center,Chinese PLA General Hospital,Beijing 100853,China;Institute of Orthopedic Medicine,the Fourth Medical Center,Chinese PLA General Hospital,Beijing 100853,China)

机构地区：[1]解放军医学院,北京100853 [2]解放军总医院第一医学中心神经外科,北京100853 [3]解放军总医院第四医学中心骨科医学研究所,北京100853

出　　处：《解放军医学院学报》2023年第6期694-699,共6页Academic Journal of Chinese PLA Medical School

基　　金：国家自然科学基金面上项目(32171356)。

摘　　要：背景施万细胞是神经系统的重要构成部分,为促进轴突再生和诱导靶神经再生提供必要的信号和空间线索。脂肪间充质干细胞来源广泛,在再生医学方向效果明确,外泌体作为神经系统细胞间的重要信息媒介,在调节再生方面发挥着重要作用。目的探索脂肪间充质干细胞来源外泌体(adipose-derived stem cells-derived exosomes,ADSCs-exos)对施万细胞增殖功能及早期神经再生特征的影响。方法提取ADSCs-exos进行鉴定;利用显微镜和流式细胞学对ADSCs鉴定,通过超速离心法提取ADSCs来源外泌体并通过透射电镜拍摄图片;将ADSCs-exos与大鼠施万细胞进行直接共培养,观察ADSCs-exos对施万细胞增殖的影响;取雌性SD大鼠8只,Matrigel基质胶搭载ADSCs-exos注入商用硅胶管构建神经移植物为外泌体组,Matrigel搭载PBS注入商用硅胶管为空导管组(Hollow),每组4只。构建坐骨神经1 cm缺损模型,将神经移植物缝合于近端和远端神经残端之间,缝合伤口后放回笼中安置。3周取出神经移植物评价轴突再生情况。结果成功提取ADSCs,能够表达间充质干细胞表面标记蛋白CD73、CD90、CD105。成功提取ADSCs-exos,能在透射电镜下观察到磷脂双分子层结构。在ADSCs-exos和施万细胞共培养体系中,比较24 h和48 h施万细胞增殖情况,施万细胞增殖与ADSCsexos浓度呈剂量依赖性。动物实验结果表明,外泌体组3周时轴突生长优于Hollow组(P<0.05)。结论ADSCs-exos可以促进施万细胞增殖,负载ADSCs-exo神经移植物的轴突生长长度在3周时即具有优势。Background Schwann cells are important components of the nervous system,providing necessary signals and spatial cues to promote axonal regeneration and induce target nerve regeneration.Adipose mesenchymal stem cells are widely sourced and have clear effects in the direction of regenerative medicine,and exosomes play an important role in regulating regeneration as an important mediator of information between cells of the nervous system.Objective To explore the effect of adipose-derived stem cells-derived exosomes(ADSCs-exos)on the function and early neural regeneration characteristics of Schwann cells.Methods ADSCs-exos were extracted for identification;ADSCs were identified by microscopic observation and flow cytometry,and ADSCs-derived exosomes were extracted by ultracentrifugation and photographed by transmission electron microscopy;ADSCs-exos were co-cultured directly with rat Schwann cells to observe the effect of ADSCs-exos on the proliferation of Schwann cells.Eight female SD rats were taken,and matrigel matrix gel was injected into commercial silicone tubes with ADSC-exos to construct nerve grafts for the exosome group;matrigel was injected into commercial silicone tubes with PBS for the empty catheter group(Hollow),with 4 rats in each group.The 1 cm sciatic nerve defect model was constructed,and the nerve grafts were sutured between the proximal and distal nerve stumps and placed back in the cage after suturing the wound.At 3 weeks after removal of the nerve grafts,the axonal regeneration was evaluated.Results ADSCs were successfully extracted and were able to express the MSC surface marker proteins CD73,CD90,and CD105.ADSCs-exos were successfully extracted and the phospholipid bilayer structure could be observed under transmission electron microscopy.In the co-culture system of ADSCs-exos and Schwann cells,the proliferation of Schwann cells at 24h and 48h was compared,and the proliferation of Schwann cells showed a significant dose-dependent relationship with the concentration of ADSC-exos.The results of

关 键 词：外周神经损伤 脂肪来源干细胞 外泌体 修复 施万细胞 

分 类 号：R651[医药卫生—外科学]