滇南小耳猪自体跟腱移植前交叉韧带重建模型的构建  

Constructing a model of anterior cruciate ligament reconstruction with autologous Achilles tendon in southern Yunnan small-ear pigs

作　　者：熊波涵 余洋[1] 郑礼玲 杨腾云 卢晓君 王旭[3] 李开炜 余鸿 李雅娟 董开颜 张瑶璋 刘津瑞 谷梓铭 胡必庚 李彦林[1] Xiong Bohan;Yu Yang;Zheng Liling;Yang Tengyun;Lu Xiaojun;Wang Xu;Li Kaiwei;Yu Hong;Li Yajuan;Dong Kaiyan;Zhang Yaozhang;Liu Jinrui;Gu Ziming;Hu Bigeng;Li Yanlin(Department of Sports Medicine,The First Affiliated Hospital of Kunming Medical University,Kunming 650032,Yunnan Province,China;Department of Clini­cal Laboratory,Quxian Hospital of Traditional Chinese Medicine,Quxian 653200,Sichuan Province,China;Department of Orthopedics,the Second Affiliated Hospital of Kunming Medical University,Kunming 650032,Yunnan Province,China;Department of Cardiology,West China Hospital,Sichuan University,Cheng­du 610000,Sichuan Province,China)

机构地区：[1]昆明医科大学第一附属医院运动医学科,云南省昆明市650032 [2]四川省达州市渠县中医院检验科,四川省渠县653200 [3]昆明医科大学第二附属医院骨科,云南省昆明市650032 [4]四川大学华西医院心内科,四川省成都市610000

出　　处：《中国组织工程研究》2024年第20期3157-3163,共7页Chinese Journal of Tissue Engineering Research

基　　金：云南省重大科技专项项目计划项目(202102AA100015),项目负责人:李彦林;云南省骨关节疾病临床医学中心项目(ZX2019-03-04),项目负责人:李彦林;云南省卫生和计划生育委员会医学领军人才培养计划任务书(L-201601),项目负责人:李彦林。

摘　　要：背景:目前关于前交叉韧带重建术的大动物模型仍较为缺乏,而滇南小耳猪作为实验动物广泛用于其他学科基础研究,但仍未见将其用于前交叉韧带重建造模的相关报道。目的:建立自体跟腱移植前交叉韧带重建的大型动物滇南小耳猪模型。方法:选取20只成年雌性滇南小耳猪,随机平均分为自体跟腱组和假手术组。自体跟腱组取小耳猪自体跟腱作为移植物行右膝关节前交叉韧带重建术,假手术组小耳猪在右膝采取类似的操作,但对前交叉韧带不行任何处理。术前及术后12个月,分别观察小耳猪一般情况;对韧带及移植物进行大体观察及MAS评分;苏木精-伊红染色观察韧带形态学特点;免疫组化评估Ⅰ型胶原与Ⅲ型胶原染色与排列情况;透射电镜观察韧带胶原纤维的类型、大小、直径、比例和分布。结果与结论:①所有动物饮食活动均正常,伤口愈合良好,无明显炎症反应,局部无化脓性感染,精神及尿便情况较术前无明显改变;膝关节重建的交叉韧带连续性完整,关节无僵硬、活动度正常,前抽屉及Lachman试验均阴性。②移植物大体观察:术后12个月关节腔内移植物位置良好,完整性尚可,张力较明显,韧带色泽接近原有前交叉韧带,表面滑膜覆盖较为完整。③自体跟腱组术后12个月关节腔内韧带大部为MASⅠ型,少数为MASⅡ型;假手术组关节腔内韧带为MASⅠ型。④苏木精-伊红染色提示,术后12个月,自体跟腱组胶原纤维已开始出现束状、同向、均匀排列,同向波纹已较为明显,细胞核以线形或纺锤形为主,与假手术组正常前交叉韧带组织较相似。⑤免疫组化提示,术后12个月,自体跟腱组重建的前交叉韧带中有较大量Ⅰ型胶原表达,Ⅲ型胶原表达明显较少;两组Ⅰ型、Ⅲ型胶原染色程度相近。⑥透射电镜显示,术后12个月,自体跟腱组重建前交叉韧带中胶原纤维直径、排列、密度与原有BACKGROUND:As a dominant breed pig in southwest China,the southern Yunnan small-ear pig has been widely used as an experimental animal in the basic research of other disciplines,but there are still no reports on its application in anterior cruciate ligament reconstruction.OBJECTIVE:To establish a large animal model of the southern Yunnan small-ear pig with anterior cruciate ligament with autologous Achilles tendon was established.METHODS:Twenty adult female Yunnan small-ear pigs were equally randomized into two groups.In the autologous Achilles tendon group,the right knee anterior cruciate ligament was reconstructed with autologous Achilles tendon as a graft,while in the sham-operated group,a similar operation was performed on the right knee without any treatment of the anterior cruciate ligament.General conditions of each pig were observed and recorded before and 12 months after surgery.Ligaments and grafts were taken for gross observation and MAS scoring.Hematoxylin-eosin staining was performed to observe morphological characteristics of ligaments.The staining and arrangement of type I and type III collagen were evaluated by immunohistochemistry.Transmission electron microscopy was used to observe the type,size,diameter,ratio,and distribution of collagen fibers in ligaments.RESULTS AND CONCLUSION:All animals had normal diet and activity,good wound healing,no obvious inflammatory reaction,no local purulent infection,and no significant changes in mental and urinary conditions compared with those before surgery.The reconstructed cruciate ligament of the knee was intact,with no stiffness and normal range of motion.Both the anterior drawer and Lachman tests were negative.Gross observation of the graft:12 months after surgery,the grafts was in good position,with good integrity,obvious tension,ligament color close to the original anterior cruciate ligament,and complete surface synovial coverage.Most of the intraarticular ligaments in the autologous Achilles tendon group were defined as MAS I type and a few were define

关 键 词：前交叉韧带 动物模型 免疫组化 透射电镜 超微结构 韧带化 

分 类 号：R496[医药卫生—康复医学] R318[医药卫生—临床医学] R-332