小肠黏膜下基质复合口腔黏膜细胞和转染TIMP-1 siRNA的成纤维细胞用于尿道重建的实验研究  

Experimental study of small intestinal submucosa seeded with oral keratinocytes and TIMP-1 siRNA transfected fibroblasts for urethral reconstruction

作　　者：郭海林[1] 撒应龙[1] 黄建文[1] 王周[1] 王林[1] 谢敏凯[1] 吕向国[1] 

机构地区：[1]上海交通大学附属第六人民医院泌尿外科,上海200233

出　　处：《临床泌尿外科杂志》2016年第3期264-269,共6页Journal of Clinical Urology

摘　　要：目的:探索应用小肠黏膜下基质(small intestinal submucosa,SIS)复合口腔黏膜细胞(oral keratinocytes,OK)和转染金属蛋白酶抑制剂-1(tissue inhibitor of metalloproteinase-1,TIMP-1)siRNA的成纤维细胞(fibroblasts,FB)构建组织工程化尿道的可行性。方法:分离培养OK,FB,并用TIMP-1siRNA转染FB,检测转染前后FB分泌Ⅰ型胶原的改变。剥离24只雄性兔子腹侧尿道黏膜2.0cm×0.8cm,并将其平均分为3组,每组8只。第1组应用单纯SIS修复,第2组应用SIS复合OK修复,第3组应用SIS复合OK和转染TIMP-1siRNA的FB修复。术后1个月、6个月行逆行尿道造影检查和组织学分析评估三组尿道重建的效果。结果:转染TIMP-1siRNA的FB分泌Ⅰ型胶原明显减少。逆行尿道造影下,第1组兔子中有5只尿道管腔通畅,第2组有6只,第3组有7只。组织学上,第1组在1个月时形成了不完整的尿路上皮,在6个月时纤维化和炎症较明显。第2组和第3组在1个月时形成了完整的尿路上皮,在6个月时纤维化和炎症较轻,且第3组生长的尿路上皮、平滑肌和血管较第2组明显增多。结论:OK和转染TIMP-1siRNA的FB可作为尿道组织工程中的种子细胞用于尿道修复重建,OK和转染TIMP-1siRNA的FB可抑制尿道瘢痕的产生。Objective：To investigate the feasibility of urethral reconstruction using small intestinal submucosa（SIS）seeded with oral keratinocytes（OK）and tissue inhibitor of metalloproteinase-1small interfering RNA（TIMP-1siRNA）transfected fibroblasts（FB）in rabbit models.Method：OK and FB were isolated and cultured.TIMP-1siRNA was transfected into FB to detect the changes of the secretion of typeⅠcollagen.In 24 male rabbits,a ventral urethral mucosal defect（2.0cm×0.8cm）was created.Urethroplasty was performed with SIS alone（eight rabbits,group one）,autogenic OK-seeded SIS（eight rabbits,group two）and autogenic OK and TIMP-1siRNA transfected FB-seeded SIS（eight rabbits,group three）.At one and six months after surgery（four rabbits at each time point）,retrograde urethrogram and histologic analysis were performed to evaluate the results of urethroplasty.Result：TIMP-1siRNA transfected FB decreased the secretion of typeⅠcollagen.Under retrograde urethrography,five rabbits in group one,six in group two and seven in group three maintained a wide urethral caliber.Histologically,the discontinued epidermal layer developed on the lumen surface of retrieval urethra at one month and inflammation and fibrosis were observed at six months in group one.However,the lumen surface formed intact urothelium at one month and mild inflammation and fibrosis were seen at six months in groups two and three.Furthermore,the speed of urothelium,smooth muscle and vessel regeneration in group three was faster than that in group two.Conclusion：OK and TIMP-1siRNA transfected FB could be used as a source of seed cells for urethral tissue engineering and could prevent the proliferation of urethral scar tissue.

关 键 词：组织工程 金属蛋白酶抑制剂-1 小肠黏膜下基质 口腔黏膜细胞 成纤维细胞 尿道重建 

分 类 号：R318[医药卫生—生物医学工程] R69[医药卫生—基础医学]