韧带后移植物移植物血供教材.docx

1、韧带后移植物移植物血供教材目 录中文摘要2英文摘要4研究论文 应用自体和异体半腱肌重建兔前叉韧带后移植物血供的实验研究前言8材料与方法10结果19附图、附表23讨论29参考文献 35综述40致谢58个人简历59应用自体和异体半腱肌重建兔前叉韧带后移植物血供的实验研究硕士研究生： 郝建学 导 师： 高石军教授 河北医科大学第三附属医院 骨科摘 要实验目的 观察兔自体和同种异体半腱肌重建前交叉韧带（anterior cruciate ligament，ACL）的组织学和组织化学变化。探讨自体和同种异体移植物重建ACL塑性改建过程以及血管生成的区别。 实验方法 24只骨骼成熟新西兰兔分为两组，分别接

2、受自体半腱肌和同种异体半腱肌重建ACL，异体半腱肌采用-80储存方法，分别于术后3周、6周、12周、24周取材，肉眼观察关节腔内重建韧带的大体形态，HE、甲苯胺兰及免疫组织化学染色观察关节腔内重建韧带的组织学变化过程及血管生成情况。结果 解剖及组织学方面：兔交叉韧带在大体解剖形态上与人类基本相同，重建后韧带在关节内经历了再血管化、细胞长入、胶原纤维再生和再塑型的过程。组织学染色显示术后3周时，组织内部呈坏死状态，周边细胞开始长入，以炎性细胞为主。甲苯胺蓝染色无异染物质出现。免疫组化血管阳性染色位于移植物韧带束膜，自体移植物组和异体移植物组无明显差别。术后6周自体移植物组进入供体的细胞和血管多于

3、异体移植物组，两组甲苯胺蓝染色无异染物质出现。术后12周大量细胞进入移植物，自体移植物组细胞数目多于异体移植物组，自体移植物组甲苯胺蓝异染物质开始出现，两组的免疫组化血管阳性染色均增加，但异体移植物组仍然少于自体移植物组。术后24周自体移植物组细胞较12周时减少，但仍比正常ACL数目多，细胞呈较统一的梭形，周围区细胞核主要为纺锤体状、杆状，中心区圆形、椭圆形细胞核稍多，可见串珠样排列，核周围有陷窝，类似于软骨细胞；胶原纤维纵向排列规律有序，并呈波状，其组织学形态已经类似正常ACL。异体移植物组细胞数目比自体移植物组多，胶原纤维束沿应力方向排列、较规律。两组甲苯胺兰染色均可以见到异染。异体移植物

4、组血管阳性染色较12周时减少，但多于自体移植物组，自体移植物组阳性染色较前减少。组织学和组织化学变化过程说明异体移植物组塑性改建过程滞后于自体移植物组。结论 正常ACL在组织形态学、组织化学和胶原分布上均表现出其特异性，与一般的肌腱和韧带组织不同，具有类软骨特性。关节腔内用来重建ACL的自体和异体半腱肌腱移植物均经历了缺血坏死炎症期、再血管化过程、成纤维细胞增殖和植入期以及韧带的塑型改造期。对于ACL重建，异体移植物的塑性改建过程滞后于自体移植物，提示临床上ACL重建术后负重时间应根据移植物类型的不同制定不同的时间。关键词: 兔；前交叉韧带；重建；自体移植物；同种异体移植物；半腱肌The Ex

5、perimental Research of Anterior Cruciate Ligament Grafts by Revascularization: Comparison of Semitendinosus Tendon Autografs and Fresh-Frozen Free-Tendon Allografts in Rabbits MM candidate: Hao JianxueSupervisor: Prof. Gao shijun The Third hospital of Hebei medical universityAbstractObjective The ob

6、jective of this research was to investigate histological and immunohistochemical changes of a free semitendinosus tendon autografs and compare these to an identical nonsterilized fresh frozen allograft for anterior cruciate ligament(ACL) reconstruction in an in vivo rabbit model and estimate the dif

7、ference in remodeling and revascularization between autografts and allografts in ACL reconstruction. Methods Twenty-four skeletally mature New Zealand White rabbits received either a semitendinosus tendon autograf or fresh frozen allograft for ACL reconstruction, the semitendinosus tendon allografs

8、were harvested from rabbits and immediately sotred in sterile bags at -80 for later use. Specimens were collected at 3, 6, 12 and 24 weeks after surgery. The gross morphology of the intraarticular segment of the grafts was observed and the remolding and revascularization of the grafts were studied b

9、y haematoxylin-eosin, toluidine blue and immunohistochemical staining. Results Anatomy and Histology: The gross anatomy of rabbit cruciate ligaments was similar to that of the human being. After implantation, the allograft tissue underwent a gradual healing process including revascularization, cellu

10、lar migration, collagen formation and remodeling. According to HE staining, intraarticular graft necrosis was evident 3 weeks after surgery. In the periphery of the grafts, host cells had already invaded into both autografts and allografts, most of them were inflammatory cells. In addition, the tolu

11、idine blue staining negative material was not observed in the both autografts and allografts. According to immunohistochemical staining, the intraarticular grafts after surgery were mainly negative on the synovium of the grafts. There wasnt found significantly different between se autografs and allo

12、grafts. At 6 weeks, allografts had a significantly lower overall cellularity and vascularity than autografts. the toluidine blue staining negative material was still not observed in the both autografts and allografts. At 12 weeks, more fibroblasts infiltrated into the allografts and autografts. Auto

13、grafts had a higher overall cellularity than allografts. In addition, the toluidine blue staining negative material was now observed in some focal concentrations in the autografts. Vascularity become more homogenous in both allografts and autografts ACL reconstructions, while vascular density in the

14、 allografts was remained lower than in the autografts. At 12 weeks, cellularity in the autografts had decreased compared to 12 weeks, but was still higher than in the intact ACL. Cellular morphology displayed a unified fusiform form. In the periphery, cell nucleus were spindle or band-shaped. Howeve

15、r, in the central portion, cell nucleus were mainly ovoid or round-shaped. Cartilage lacuna-like structure was observed. the stress-oriented collagen fibers in the autograft were similar to the normal ligament. Cellularity in the allografts was higher than that in the autografts. The collagen fibers in the allografts aligned on the stress direction. The toluidine blue staining negative