组织工程尿道粘膜的体外构建

Construction of Urethra Mucosa Structure in Vitro by Tissue Engineering

目的:利用组织工程方法在体外构建形成尿道粘膜结构。方法:酶消化法获得猪膀胱尿路上皮细胞(UC),以免疫荧光和RT-PCR方法进行UC鉴定。制备膀胱脱细胞基质移植物(BAMG)作为支架材料,以苏木精-伊红染色、Masson染色、免疫组化和扫描电镜进行评价。经过体外培养和扩增后,将P3代UC接种在BAMG上。结果:BAMG支架上的UC经过体外培养1周后即可形成沿基膜排列的复层上皮结构。扫描电镜显示细胞-材料复合良好,免疫组化检测支架上的UC广谱角蛋白表达阳性。结论:运用组织工程方法能够在体外快速进行尿道粘膜上皮结构的初步构建,为进一步临床修复诸如尿道下裂、尿道狭窄等尿道缺损奠定技术基础。

Objective ： To construct a structure of urethra mucosa in vitro by tissue engineering. Methods ： Primary porcine urothelial cells （UC） were obtained from the porcine bladder by enzymatic digestion and detected by immunofluorescence and RT-PCR. Bladder acellular matrix grafts （BAMG） were prepared, used as the scaffold and then evaluated by HE staining, Masson＇s trichrome staining, immunohistology and scanning electron microscopy. After in vitro culture and amplification, the UCs were seeded on the luminal surface the BAMGs. Results ： After 1 week of in vitro culture, the UCs formed a muhilayer structure on the luminal surface of the BAMGs along the basement membrane. The tissue-engineered urothelium and BAMG complex was well formed and pan cytokeratins were positively expressed in the UCs on the scaffold. Conclusion ： By tissue engineering, the urethra mucosa structure can be rapidly constructed in vitro, which can be applied to the repair of such urethral defect as hypospadias and urethral stricture.