富血小板血浆源性外泌体对肌腱干/祖细胞增殖及迁移能力的影响  

Effects of Platelet-Rich Plasma-Derived Exosomes on Proliferation and Migration of Tendon Stem/Progenitor Cell

作　　者：李墨琳 朱亚琼 丁雨菲 易丹 葛乃侨 陈思明 王月香 LI Molin;ZHU Yaqiong;DING Yufei;YI Dan;GE Naiqiao;CHEN Siming;WANG Yuexiang(Chinese PLA Medical School,Beijing 100853,China;Department of Ultrasound,The First Medical Center of Chinese PLA General Hospital,Beijing 100853,China;Department of Orthopedics,Beijing Friendship Hospital,Capital Medical University,Beijing 100050,China;Department of Radiology,Armed Police Corps Hospital of Jilin Province,Jilin,Jilin 130000,China)

机构地区：[1]解放军医学院,北京100853 [2]中国人民解放军总医院第一医学中心超声诊断科,北京100853 [3]首都医科大学附属北京友谊医院骨科,北京100050 [4]武警吉林省总队医院医学影像科,吉林吉林130000

出　　处：《中国医学科学院学报》2024年第3期307-315,共9页Acta Academiae Medicinae Sinicae

基　　金：国家自然科学基金(82102079)。

摘　　要：目的探究富血小板血浆源性外泌体(PRP-Exos)对肌腱干/祖细胞(TSPC)增殖及迁移能力的影响。方法采用聚合沉淀结合超速离心法提取PRP-Exos,采用透射电镜和纳米颗粒跟踪分析技术鉴定其形态、浓度及粒径,采用蛋白免疫印迹法检测外泌体表面标志蛋白及血小板膜糖蛋白的表达水平。提取并培养大鼠TSPC,采用流式细胞术及免疫荧光染色检测TSPC表面标志分子的表达情况。CCK-8法及EdU实验评估PRP-Exos对TSPC增殖情况的影响。细胞划痕实验及Transwell实验评估PRP-Exos对TSPC迁移情况的影响。结果提取的PRP-Exos呈典型茶托状结构,浓度为4.9×10^(11)个/mL,平均粒径为(132.2±56.8)nm,PRP-Exos表面表达CD9、CD63及CD41。提取的TSPC表达CD44蛋白。PRP-Exos可被TSPC摄取,共培养48 h时,浓度为50、100μg/mL的PRP-Exos可显著促进TSPC的增殖(P均<0.001),且两组之间差异无统计学意义(P=0.283);共培养24 h后,浓度为50μg/mL的PRP-Exos可显著促进TSPC的迁移(P<0.001)。结论体外培养条件下,PRP-Exos对大鼠TSPC的增殖及迁移具有显著促进作用。Objective To investigate the effects of platelet-rich plasma-derived exosomes(PRP-Exos)on the proliferation and migration of tendon stem/progenitor cell(TSPC).Methods PRP-Exos were extracted through the combination of polymer-based precipitation and ultracentrifugation.The morphology,concentration,and particle size of PRP-Exos were identified by transmission electron microscopy and nanoparticle tracking analysis.The expression levels of surface marker proteins on PRP-Exos and platelet membrane glycoproteins were determined by Western blot analysis.Rat TSPC was extracted and cultured,and the expression of surface marker molecules on TSPC was detected using flow cytometry and immunofluorescence staining.The proliferation of TSPC influenced by PRP-Exos was evaluated using CCK-8 assay and EdU assay.The effect of PRP-Exos on the migration of TSPC was evaluated by cell scratch assay and Transwell assay.Results The extracted PRP-Exos exhibit typical saucer-like structures,with a concentration of 4.9×10^(11)particles/mL,an average particle size of(132.2±56.8)nm,and surface expression of CD9,CD63 and CD41.The extracted TSPC expressed the CD44 protein.PRP-Exos can be taken up by TSPC,and after co-cultured for 48 h,concentrations of 50 and 100μg/mL of PRP-Exos significantly promoted the proliferation of TSPC(both P<0.001),with no statistical difference between the two concentrations(P=0.283).Additionally,after co-cultured for 24 h,50μg/mL of PRP-Exos significantly promoted the migration of TSPC(P<0.001).Conclusion Under in vitro culture conditions,PRP-Exos significantly promote the proliferation and migration of rat TSPC.

关 键 词：富血小板血浆 外泌体 肌腱干/祖细胞 增殖 迁移 肩袖损伤 

分 类 号：R459.9[医药卫生—治疗学] R686.5[医药卫生—临床医学]