干细胞与膀胱脱细胞基质构建组织工程管状移植物修复长段输尿管缺损  

Tissue-engineered tubular grafts constructed with stem cells and bladder acellular matrix in long-segmental ureteral reconstruction

作　　者：廖文彪[1] 杨嗣星[1] 熊云鹤[1] 宋超[1] 孟令超[1] 

机构地区：[1]武汉大学人民医院泌尿外科二科,湖北省武汉市430060

出　　处：《中国组织工程研究》2015年第10期1570-1574,共5页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(31400835)~~

摘　　要：背景:临床上重建长段输尿管缺损的方法存在并发症多且创伤大等缺点,因此,需要寻找其他的修复方式。目的:探讨组织工程管状移植物修复长段输尿管缺损的可行性。方法:将兔骨髓间充质干细胞和膀胱平滑肌细胞种植到兔膀胱脱细胞基质的两面,培养7 d后制作成4 cm长的组织工程管状移植物,仅将膀胱平滑肌细胞种植到兔膀胱脱细胞基质构建组织工程管状移植物作为对照;新西兰兔25只,随机分为实验组20只和对照组5只,将2种组织工程管状移植物移植到兔体内修复输尿管缺损,修复同时利用网膜包裹,分别于术后2,4,8周取实验组标本做苏木精-伊红染色及免疫组化检测。结果与结论:组织学检查发现实验组尿路上皮层再生呈连续性,术后8周可见组织工程管状移植物管腔内覆盖多层尿路上皮结构及规则的平滑肌束,免疫组化可见uroplakinⅢa及CK AE1/AE3阳性,证实了组织工程管状移植物组织内有成熟上皮细胞覆盖,且上皮细胞之间有连接功能。对照组5只动物术后拔除输尿管支架管2周内均死亡,尸检见流出道瘢痕形成,组织工程管状移植物内腔闭锁,同侧肾脏重度积水,未作组织学观察。以上结果表明将骨髓间充质干细胞和膀胱平滑肌细胞种植于兔膀胱脱细胞基质上构建组织工程管状移植物修复长段输尿管缺损是可行的,组织工程管状移植物内腔有成熟上皮覆盖,骨髓间充质干细胞能促进尿路上皮再生。BACKGROUND： The main way for long-segmental ureteral reconstruction may cause a lot of traumas and complications. Therefore, to seek a new repair method is urgent.OBJECTIVE： To investigate the feasibility of a tissue-engineered tubular graft for ureteral reconstruction.METHODS： Bone marrow mesenchymal stem cells and smooth muscle cells of rabbits were seeded into the two surfaces of bladder acellular matrix and cultivated for 7 days. Then the graft was used to prepare a 4-cm long tissue-engineered tubular graft, which was regarded as experimental group. Smooth muscle cells seeded onto the bladder acellular matrix was used to construct the tissue-engineered tubular graft as control group. Twenty-five New Zealand rabbits were randomly divided into experimental group（n=20） and control group（n=5）, and two kinds of tubular grafts covered with omentum were implanted into the two groups, respectively, for repair of ureteral defects. Hematoxylin-eosin staining and immunohistochemical detection were performed at 2, 4, 8weeks after implantation.RESULTS AND CONCLUSION： In the experimental group, hematoxylin-eosin staining showed epithelial coverage and muscle fibers on the lumen of tissue-engineered tubular grafts at 8 weeks after implantation;immunohistochemistry showed that anti-AE1/AE3 antibody and anti-uroplakin Ⅲa antibody were positive,confirming that there were mature epithelial cells on the lumen of tissue-engineered tubular grafts. In the control group, five rabbits were dead within 2 weeks after removal of ureteral scaffold, and autopsy showed scar formation inside the graft and severe hydronephrosis. These results demonstrate that it is feasible to construct the tissue-engineered tubular graft using bone marrow mesenchymal stem cells and smooth muscle cells into the bladder acellular matrix for ureteral reconstruction. Bone marrow mesenchymal stem cells can potentially promote urothelial regeneration.

关 键 词：骨髓 间质干细胞 肌细胞 平滑肌 输尿管疾病 干细胞 移植 膀胱脱细胞基质 骨髓间充质干细胞 平滑肌细胞 输尿管缺损 尿路上皮再生 组织工程 国家自然科学基金 

分 类 号：R394.2[医药卫生—医学遗传学]