人脐带间充质干细胞联合富含血小板血浆促进大鼠跟腱损伤的修复

发布时间：2018-04-21 06:55

  本文选题：人脐带间充质干细胞 + 富含血小板血浆　； 参考：《第三军医大学学报》2017年02期

【摘要】：目的探讨不同浓度富含血小板血浆(platelet-rich plasma,PRP)在体外对人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,h UC-MSCs)增殖、分化的影响,评估h UC-MSCs联合PRP对大鼠跟腱损伤修复的影响。方法 PRP活化后成为PRP-clot releasate(PRCR)。取P3代h UC-MSCs于培养皿中培养24 h后,分别加入含0%(对照组)、10%、20%、40%PRCR的DMEM/F12培养基,于第1、2、3、4天测定h UC-MSCs的增殖速度。P3代h UC-MSCs按相同分组培养,于第1、3、5天收集细胞并提取蛋白。Western blot检测h UC-MSCs中成肌腱分化标志性基因肌腱蛋白C(tenascin c,TNC)、SCX(scleraxis)、Ⅰ型胶原的表达。选取48只8周龄雄性SD大鼠,Ⅰ型胶原酶溶液50μL注射于大鼠右侧跟腱制备跟腱损伤模型,3 d后于损伤部位再次分别注射50μL生理盐水(对照)、h UC-MSCs悬液、PRP或PRP+h UC-MSCs混悬液,术后2、4周安乐死大鼠,Western blot检测跟腱中TNC、SCX及Ⅰ型胶原的表达,跟腱切片HE染色行组织学评估。结果 PRCR在体外对h UC-MSCs增殖均有促进作用,20%PRCR组促增殖作用最显著(P0.05);20%PRCR组较对照组在体外可以显著地促进h UCMSCs成肌腱分化标志性基因的表达(P0.05);术后2、4周,PRP+h UC-MSCs组跟腱中成肌腱分化相关基因在蛋白水平表达更高,跟腱切片HE染色显示PRP+h UC-MSCs组纤维排列更规整,炎细胞更少。结论适宜浓度PRP在体外显著促进了h UC-MSCs的增殖、成肌腱分化;PRP联合h UC-MSCs能有效促进大鼠受损跟腱的修复。
[Abstract]:Objective to investigate the effects of platelet-rich plasma-rich plasma with different concentrations of platelet-rich plasma on proliferation and differentiation of human umbilical cord mesenchymal stem cells hUC-MSCs in vitro, and to evaluate the effects of h UC-MSCs combined with PRP on the repair of Achilles tendon injury in rats. Methods PRP was activated to become PRP-clot release. After cultured in culture dish for 24 hours, P3 generation h UC-MSCs was added into DMEM/F12 medium containing 0 (control group 10) and 20% 40 PRCR respectively. The proliferation rate of h UC-MSCs. P3 generation h UC-MSCs were cultured according to the same group on day 1, 2, 3 and 3, 4 days later. The cells were collected on the 3rd day of the first trimester, and the protein was extracted. Western blot was used to detect the expression of the tendon protein (C(tenascin), a typical gene of tendon differentiation in h UC-MSCs, SCX scleraxisus, type 鈪，

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