大网膜包埋移植肝脏去细胞生物支架血管化过程观察  

Observation on the process of vascular reconstruction in acellular hepatic scaffolds of rat after transplantation into the greater omentum

作　　者：黄俊杰[1,2] 王志斌 陈纳[3] 张建色 王志翊[1] 黎婷[1] 戴开宇[1] 梅劲[1] 楼新法[1] HUANG Junjie;WANG Zhibin;CHEN Na;ZHANG Jianse;WANG Zhiyi;LI Ting;DAI Kaiyu;MEI Jin;LOU Xinfa(Institute of Bioscaffold Transplantation and Immunol-ogy, Wenzhou Medical University, Wenzhou, 325035;School of the Second Clinical Medical Sciences, Wenzhou Medical University, Wenzhou, 325035;Department of General Surgery, the First Affliated Hospital of Zhejiang University, Hangzhou, 310058)

机构地区：[1]温州医科大学生物支架移植与免疫研究所,浙江温州325035 [2]浙江大学附属第一医院普外科,浙江杭州310058 [3]温州医科大学第二临床医学院,浙江温州325035

出　　处：《温州医科大学学报》2017年第10期703-707,共5页Journal of Wenzhou Medical University

基　　金：浙江省自然科学基金资助项目(LY13H030010,LY14H180008);浙江省医药卫生科技计划项目(2013KYA121,2016KYA061,2016KYA135);温州市科技局公益类项目(Y20140664);浙江省大学生科技创新活动计划项目(2014R413024,2015R413031)

摘　　要：目的:观察肝脏去细胞生物支架(AHS)大网膜包埋移植后的血管新生过程。方法:采用Triton X-100灌注法制备大鼠肝脏AHS,HE染色、扫描电镜、铸型、DNA含量检测、免疫荧光染色等方法鉴定去细胞效果;通过大网膜包埋移植肝脏AHS,分别于术后1、3、7、14、28d处死大鼠,病理切片HE染色观察移植肝脏AHS血管新生过程。结果:灌注法制备的肝脏AHS空间结构与细胞外基质成分保留完整,细胞成分基本被去除,AHS组DNA含量与正常对照组比差异有统计学意义(P<0.01);大网膜包埋移植肝脏AHS术后3d,靠近网膜移植区的支架内出现血窦,术后7d出现新生血管,并逐渐向移植支架中心区域延伸,血管逐步分化成熟,14d以后形成血管化网状组织,28d以后血管化网状组织仍存在。结论:大网膜包埋移植促使肝脏AHS再细胞化,最终可形成血管化的网状结构。Objective： To observe the process of vascular reconstruction in acellular hepatic scaffolds of rat after transplantation into the greater omentum. Methods： Acellular hepatic scaffolds were harvested from adult SD rats by perfusing with Triton X-100. The scaffolds and native livers were observed through genomic DNA content analysis, scanning electron microscopy, HE stain, immunofluorescence and vascular cast. Then the acellular hepatic scaffolds were transplanted into the greater omentum in rats to investigate the vascular reconstruction. Rats were sacrificed respectively at 1, 3, 7, 14 or 28 days post-implantation and sections from the explanted grafts were analyzed by HE stain for morphology and microstructures. Results： Scanning electron microscope, HE stain and immunofluorescence showed most collagen fibers but no visible cell nuclei remained after decellularization in the scaffolds compared with the native livers. Vascular cast showed the pipeline structures of acellular hepatic scaffolds were intact and clear. Quantitative analysis of DNA content within the scaffolds showed only（211.6 ± 25.3） ng/μL. Then the blood sinus were observed on border of grafts at 3 days after transplantation into greater omentum. Afterwards the neovessels appeared, got into the internal of scaffolds, and formed microvascular network gradually in hepatic scaffolds after 14 days. The de novo microvascular network were still existed in hepatic scaffolds on 28 days. Conclusion： The implantation of greater omentum can promote the recellularization and eventually formation of vascularized reticular structures in the decellularized hepatic scaffolds.

关 键 词：肝脏 去细胞 血管 组织工程 大鼠 

分 类 号：R622[医药卫生—整形外科]