蚕丝-胶原支架用于兔前交叉韧带重建的研究  

Anterior cruciate ligament reconstruction in a rabbit model using silk-collagen scaffold

作　　者：田科[1] 陈央娣 梁魁 李智富[1] 许建中[1] 毕方刚[1] Tian Ke;Chen Yangdi;Liang Kui;Li Zhifu;Xu Jianzhong;Bi Fanggang(Department of Orthopedic Surgery,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Henan University of Chinese Medicine,Zhengzhou 450052,China;Department of Surgery,Zhengzhou University Hospital,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院骨科,450052 [2]河南中医药大学,郑州450052 [3]郑州大学医院外科,450052

出　　处：《中华实验外科杂志》2020年第9期1691-1694,共4页Chinese Journal of Experimental Surgery

基　　金：河南省教育厅2019年重点科研项目(32211051);河南省科技厅2020年科技攻关项目(22170139)。

摘　　要：目的评估蚕丝-胶原支架用于兔前交叉韧带(ACL)重建后,关节腔内韧带再生和骨隧道内腱-骨愈合。方法Na2CO3脱胶法对生蚕丝进行脱胶处理,乙酸抽提法获取Ⅰ型胶原,然后使用热交联和冷冻干燥法制备蚕丝-胶原材料。20只12周龄新西兰大白兔(由河南省实验动物中心提供),对左膝关节使用蚕丝-胶原支架重建ACL。术后4、16周两个时间点随机处死实验动物的一半,每组标本中的5个进行组织学检测,包括关节腔内韧带的组织学检测和骨隧道腱-骨愈合的组织学检测;每组中余下5个标本进行Micro-CT和生物力学检测。组间比较采用两独立样本t检验。结果术后4周,关节腔内的蚕丝-胶原支架表面可见大量的细胞浸润,细胞排列方向杂乱,细胞周围基质较少,且材料内部细胞浸润较少;骨道内蚕丝-胶原材料内可见细胞浸润,细胞排列较为松散,腱-骨界面可见新生骨组织,骨小梁结构明显;Micro-CT显示骨隧道内新生骨较少,新生骨小梁稀疏,骨体积分数(BV/TV)=(19.36±2.29)%,骨密度(BMD)=(245.04±17.68)mg/cm3。术后16周,关节腔内支架表面和内部均可见大量细胞浸润,细胞呈纤维细胞形态,排列方向较为一致,与支架长轴方向一致,细胞周围基质较多,免疫组织化学染色显示肌腱蛋白-C(Tenascin-C)表达较4周明显增多;骨道内蚕丝-胶原材料内细胞浸润进一步增多,细胞排列较4周时更为有序,腱-骨界面骨组织趋于成熟,骨组织与支架材料结合更加紧密;Micro-CT显示骨隧道内新生骨较4周明显增多,BV/TV=(39.25±1.51)%(t=16.010,P<0.05),BMD=(400.88±58.32)mg/cm3,组间比较差异有计学意义(t=5.718,P<0.05);16周5例标本测得最大拉力[(43.67±6.52)N],刚度[(9.18±0.76)N/mm],较4周差异有统计学意义[最大拉力(25.87±4.57)N,t=4.994,P<0.05;刚度(4.85±0.84)N/mm,t=8.556,P<0.05]。结论蚕丝-胶原支架用于兔ACL重建,不仅能获得较好的关节腔内韧带再生,同时能够达到良Objective To investigate the ligament regeneration in the knee cavity and the osteointegration at the tendon-bone interface after anterior cruciate ligament(ACL)reconstruction using silk-collagen scaffold in a rabbit model.Methods The silk-collagen scaffold was fabricated using degummed knitted silk scaffold and collagenⅠmatrix.The silk-collagen scaffold was rolled up to make a graft for replacing native ACL in the rabbit model.All specimens were harvested at 4 and 16 weeks postoperatively to evaluated ligament regeneration by hematoxylin and eosin(HE)and immunohistochemistry staining,and tendon-bone healing by HE and Russell-Movat staining.Micro-CT,biomechanical tests were also used to assess the tendon-bone healing process.Results At 16 weeks postoperatively,HE and immunohistochemistry staining revealed that more aligned cell arrangement and more extracellular matrix(ECM)deposition in the silk-collagen scaffold in the knee cavity than those of 4 weeks postoperatively.At week 16,more bone formation which grew into the silk-collagen scaffold was observed,while new trabecular bone could be observed at 4 weeks after surgery.BV/TV=(39.25±1.51)%,bone mineral density(BMD)=(400.88±58.32)mg/cm3,at 16 weeks postoperatively were significantly higher than those at 4 weeks[BV/TV=(19.36±2.29)%(t=16.010,P<0.05),BMD=(245.04±17.68)mg/cm3,t=5.718,P<0.05]after surgery.Higher signals indicating more bone mineral formation were detected in the specimens of 16 weeks than those of 4 weeks postoperatively,which were consistent with the results of biomechanical test[failure load,(25.87±4.57)N vs.(43.67±6.52)N,t=4.994,P<0.05;stiffness,(4.85±0.84)N/mm vs.(9.18±0.76)N/mm,t=8.556,P<0.05].Conclusion Silk-collagen scaffold induced ligament regeneration in the knee cavity and osteointegration at the tendon-bone interface after ACL reconstruction.

关 键 词：蚕丝-胶原支架 前交叉韧带 再生 腱-骨愈合 

分 类 号：R687.4[医药卫生—骨科学] R318.08[医药卫生—外科学]