种植自体尿路上皮细胞的包膜化输尿管组织工程支架的构建.docVIP

·48·  doi 10.3969/j.issn.1673-4254.2013.01.11  J South Med Univ, 2013, 33(1): 48-52  基础研究 种植自体尿路上皮细胞的包膜化输尿管组织工程支架的构建 谭海颂 1，符伟军 1，李健强 1，王忠新 1，李 钢 2，马 鑫 1，董 隽 1，高江平 1，王晓雄 1，张 旭 1 1 解放军总医院泌尿外科，北京 100853；2解放军第 309 医院泌尿外科，北京 100091  摘要：目的 探讨应用组织工程技术体外构建种植自体尿路上皮细胞的包膜化聚乳酸输尿管支架的可行性。方法 将聚乳酸输 尿管支架于比格犬皮下包埋 3 周，在其表面诱导形成一层结缔组织薄膜，获得包膜化聚乳酸输尿管支架。经脱细胞处理后，将 体外扩增培养的自体尿路上皮细胞种植于包膜输尿管支架表面。采用常规 HE 染色组织切片、扫描电镜和免疫组化法对包膜输 尿管支架及尿路上皮附着情况进行观察；应用 MTT 比色法比较尿路上皮细胞在包膜化聚乳酸输尿管支架组和管状小肠粘膜下 基质组的生长差异。结果 HE 染色及因子免疫组化染色结果显示结缔组织层内含有大量毛细血管，但无明显局部炎症反应； 扫描电镜和免疫组化染色提示包膜含有丰富的胶原纤维构成、表面呈现三维立体结构，尿路上皮细胞能在支架包膜表面粘附， 形成连续的上皮层；MTT 检测显示包膜化聚乳酸输尿管支架组和管状小肠粘膜下基质组组吸光光度值比较无明显差异（P 0.05），说明尿路上皮细胞在包膜化输尿管支架上能连续增殖。结论 包膜化聚乳酸输尿管支架适合尿路上皮细胞黏附和增殖， 体外构建的包膜化组织工程输尿管有潜在的临床应用价值。 关键词：输尿管；支架；脱细胞基质；组织工程 Construction of a capsular tissue-engineered ureteral stent seeded with autologous urothelial cells TAN Haisong1, FU Weijun1, LI Jianqiang1, WANG Zhongxin1, LI Gang2, MA Xin1, DONG Jun1, GAO Jiangping1, WANG Xiaoxiong1, ZHANG Xu1 1 China Abstract: Objective To investigate the feasibility of constructing a capsular poly L-lactic acid (PLLA) ureteral stent seeded with autologous urothelial cells using tissue engineering methods. Methods The capsular ureteral stent was constructed by subcutaneously embedding PLLA ureteral stent in the back of beagles for 3 weeks to induce the formation of connective tissue on the surfaces. After decellularization of the stent, the expanded autologous urothelial cells were seeded on the stent. The surface structure and cell adhesion of the stent were observed using HE staining, scanning electron microscope (SEM) and immunocytochemichal staining. MTT assay was used to evaluate urothelial cell proliferation on the capsular PLLA ureteral stent and on circumferential small intestinal submucosa graft. Results HE staining and VIII factor immunohistochemistry revealed numerous capillaries in the connective tissue encapsulating the stent without obvious local inflammatory response. The results of SEM and immunocytoch