外周血间充质干细胞凝胶对兔肌腱移植物重建前交叉韧带腱骨愈合的影响  

The tendon-to-bone healing of ACL reconstruction in a rabbit model using autograft seeded with PBMSCs gel

作　　者：刘家能[1] 徐斌[1] 徐洪港[1] 朱雪坤[1] 陈鹏[1] 

机构地区：[1]安徽医科大学第一附属医院运动创伤与关节镜外科,合肥230032

出　　处：《安徽医科大学学报》2015年第8期1081-1085,共5页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金(编号:1208085MH157)

摘　　要：目的：探讨在兔自体肌腱移植物重建前交叉韧带（ACL）模型股骨道内注入自体外周血间充质干细胞（PBM-SCs）凝胶对早期腱骨愈合的影响。方法健康3～4月龄新西兰大白兔32只,随机均分为 PBMSCs 组与对照组,PBM-SCs 组予以皮下注射粒细胞集落刺激因子（G-CSF）动员6 d后采集外周血,采用密度梯度离心法及贴壁培养法分离纯化PBMSCs,光镜下观察细胞形态并行流式细胞术检测鉴定表面抗原表达。建立自体肌腱移植物重建前交叉韧带模型, PBMSCs 组于动物模型股骨道内注入 PBMSCs 凝胶,对照组仅注入凝胶。分别于术后第2、4、8、12周每组随机取4只处死取材,1只做 Masson 三色染色观察组织愈合的病理表现,3只做股骨道移植物抗牵拉试验观察其愈合强度。结果流式细胞术检测结果提示 PBMSCs 表面表达 CD29、CD90,不表达 CD11b、CD34。 Masson 三色染色显示 PBMSCs 组术后第4周胶原纤维开始增生,排列紊乱,第8周胶原纤维显著增生,排列不规则,第12周胶原纤维大量增生,排列有序。 PBM-SCs 组腱骨愈合速度较对照组迅速。 PBMSCs 组和对照组的股骨道移植物抗牵拉强度随时间延长呈现上升趋势,其中PBMSCs 组移植物的抗牵拉强度自第8周起明显高于对照组（P ＜0．01）。结论 PBMSCs 凝胶对兔自体肌腱移植物重建ACL 的早期腱骨愈合有促进作用。Objective This study was conducted to analyze the effect of peripheral blood mesenchymal stem cells （PBMSCs） gel on the early period of tendon-to-bone healing after reconstruction of the anterior cruciate ligament （ACL） using autograft. Methods Thirty-two New Zealand rabbits aged 3 ～ 4 months were randomly divided into two groups （n = 16 per group）： experimental and control. After rabbits from the experimental group were mobilized by subcutaneous injection of granulocyte colony stimulating factor （G-CSF） for 6d, the PBMSCs were isolated from rabbit peripheral blood samples by combination of density gradient centrifugation and adhesive culture, morphology of cell proliferation was observed by microscopy and phenotypes of PBMSCs were detected by flow cytometry. A model of ACL reconstruction using autograft was performed in all of the rabbits, the experimental group received in-jection of PBMSCs gel into the femur tunnel and the control group received gel only. At 2, 4, 8 and 12 weeks, 4 animals in each group were euthanized, 1 for histologic assessment by Masson trichrome staining and 3 for biome-chanical testing. Results Flow cytometry showed that the PBMSCs expressed CD29, CD90 positive, CD11b, CD34 negative. According to the Masson trichrome staining, in the experimental group, collagen fibers were pres-ented but had a disordered arrangement at 4 weeks after surgery. At 8 weeks after surgery, collagen fibers were ob-viously increased but irregularly arranged. At 12 weeks after surgery, collagen fibers were formed in abundance and regularly arranged. The healing at tendon-bone interface of the experimental group was quicker than those of the control group. The maximum biomechanical pull-out strength of the autograft in the femur tunnel presented a rising trend with the prolong of repair time in both experimental and control groups, and it was significantly higher in the experimental group at 8 and 12 weeks after surgery （P 〈 0. 01）. Conclusion PBMSCs gel could enhance the early period of

关 键 词：前交叉韧带 腱骨愈合 外周血间充质干细胞 

分 类 号：R686.5[医药卫生—骨科学]