内纵外螺旋双层结构管状尿道支架复合体修复尿道缺损的可行性及其血管化方法  

The effects of inner vertical outer spiral complex scaffold in repairing long segment of urethral defect

作　　者：杜小文[1] 陈浩浩[1] 刘庆[1] 项见阳 王巧[1] 徐挺[1] 严秋亮[1] 冯超[2] Du Xiaowen Chen Haohao Liu Qing Xiang Jianyang Wang Qiao Xu Ting Yan Qiuliang Feng Chao(Department of Urinary Surgery, Jinhua People ＇s Hospital, Zhe Jiang Province, Jinhua 321000, China)

机构地区：[1]浙江省金华市人民医院泌尿外科,321000 [2]上海交通大学附属第六人民医院泌尿外科

出　　处：《中华泌尿外科杂志》2017年第1期59-65,共7页Chinese Journal of Urology

基　　金：浙江省科技计划项目（2013C33102）

摘　　要：目的研究内纵外螺旋双层结构的管状尿道支架复合体血管化的方法，探讨采用其修复长段尿道缺损的可能性。方法2014年8月至2015年10月选取雄性新西兰大白兔附睾旁脂肪，分离培养脂肪间充质干细胞（adipose－derived mesenchymal stem cells，ADSC）及经过转化的平滑肌细胞为种子细胞，并采用重组血管内皮生长因子（vascular endothelial growth factor，VEGF）基因慢病毒转染。以小肠黏膜下层脱细胞基质和经过Ⅰ型胶原蛋白表面修饰的聚乳酸乙酸共聚物（polylactic－co－glycolic acid，PLGA ）膜为组织工程材料，构建内纵外螺旋双层结构的管状尿道支架，支架长度3.0 cm。采用一次沉淀法将转染的ADSC和平滑肌细胞分别种植于尿道支架的内外层，构建管状尿道支架复合体。抽签法将27只清洁级雄性新西兰大白兔分为转染组（S1组）、带血管蒂转移皮管组（S2组）、单纯支架组（C组），每组9只。手术切除3.0 cm尿道构建尿道缺损动物模型。S1组采用尿道支架复合体直接修复尿道缺损；S2组将未转染慢病毒的ADSC和平滑肌细胞种植尿道支架后，包埋于实验动物腹股沟皮下血管旁，3周后采用带血管蒂转移皮管修复尿道缺损；C组直接采用尿道支架修复尿道缺损。术后4、8、24周分别选取每组各3只观察记录并发症发生情况和尿道造影检查结果，处死动物后留取修复段尿道标本行形态观察、扫描电镜观察以及HE、荧光示踪和相关蛋白免疫组化染色检查。结果S1组术后尿瘘、尿道下裂及尿道狭窄相关性死亡各1只；尿道造影检查示尿道光滑通畅、粗细较均匀；病理检查见尿道黏膜修复良好，较平整光滑。S2组术后尿瘘和尿道狭窄相关性死亡分别为1只和2只；尿道造影示尿道壁粗糙、局部变细或扩张；病理检查见尿道黏膜下多量脂肪堆积，黏膜收缩。C组术后尿瘘、尿道下裂、尿道�Objective To explore the possibility of the inner vertical outer spiral complex tubular urethra scaffold vascularization in repairing long segment of urethral defect. Methods From August 2014 to October 2015,27 clean male New Zealand white rabbits were divided into 3 groups ,S1 group was transfeeted recombinant vascular endothelial growth factor（VEGF） gene lentiviral vector group. $2 group was vascular pedicle transfer tube group. C group was simple stent group. A 3.0 cm inner vertical outer spiral complex scaffold was constructed by using the small intestine aeellular matrix （SIS） and polylactic acid copolymer （PLGA） modified by type I collagen surface, and adipose-derived mesenehymal stem cells （ADSC） and smooth muscle cells after transformation from New Zealand white rabbits. In S1 group, the seed cells were transfected by recombinant vascular endothelial growth factor （VEGF） gene lentivirus, which express VEGF protein. The complex scaffold was used to repair 3.0 cm rabbit urethral defect In $2 group, the untransfected cells were seeded into the scaffold and embedded in the skin near the groin artery 3 weeks for repairing urethral defect with vascular pedicle transfer tube. In group C, the unseeded scaffold was used to repair the urethral defect alone. Postoperative observation and urethrography were followed 4, 8 and 24 weeks after implantation. The HE staining, fluorescence tracing, immunohistochemical and scanning electron microscopy were evaluated at the same phase. Results In S1 group, there were one urinary fistula and one urethral stricture-related death, respectively. The urethra was smooth and patent, histological examination showed active hyperplasia of urethral capillary. In S2 group, there were one urinary fistula and two urethral stricture-related deaths, respectively. The urethral was rough, local thinning or dilated. Fat accumulation and mucosal contraction were found in the urethral submucosal, respctively. In C group, there were one urinary fistula, three hypospadias, and t

关 键 词：组织工程 支架 尿道缺损 血管化 

分 类 号：R699[医药卫生—泌尿科学]