Collagen cell carriers seeded with human urothelial cells for urethral reconstructive surgery: first results in a xenograft minipig model
Collagen cell carriers seeded with human urothelial cells for urethral reconstructive surgery: first results in a xenograft minipig model
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DOI：
10.1007/s00345-016-1959-3
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发表时间：
2017-07-01
影响因子：
3.4
通讯作者：
Sievert, Karl-Dietrich
中科院分区：
文献类型：
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作者：
Aufderklamm, Stefan;Vaegler, Martin;Sievert, Karl-Dietrich
目的尿道狭窄是男性下尿路的常见病。目前，对于长期或复发性狭窄，使用颊粘膜是首选的方法。然而，自体组织工程移植在泌尿外科重建中的应用仍处于研究阶段。本初步研究的目的是在动物模型中评估人类尿路上皮细胞（HUC）在牛胶原i型细胞载体（CCC）上的应用，并评估手术过程的短期结果。方法采用免疫抑制剂环孢素A对4只雄性哥廷根小型猪进行初步研究。从人良性输尿管组织中获得HUC，用PKH26染色，并将其植入胶原细胞载体（CCC）。在尿道狭窄诱导和保护性膀胱造口术后7周，分别用HUC管腔和抗管腔植入细胞种子CCC。两周后对动物实施安乐死，进行尿道造影和组织学评估。结果所有小型猪手术在技术上是可行的。诱导后7周影像学证实狭窄。两周后可见CCC，并表现出良好的整合，无炎症或排斥迹象。在最后的尿道造影中，没有发现剩余的狭窄。在猪尿路上皮附近，即使与CCC部分分离，也发现pkh26阳性区域。pankeratin, CK20， E-cadherin和ZO-1的免疫荧光显示在CCC上和附近的几个区域尿路上皮完整。最后，本研究表明huc种子的CCC作为小型猪的异种移植物在技术上是可行的，并且具有进一步研究的前景。
Purpose Urethral strictures are a common disease of the lower urinary tract in men. At present, the use of buccal mucosa is the method of choice for long or recurrent strictures. However, autologous tissue-engineered grafts are still under investigation for reconstructive urological surgery. The aim of this pilot study was to evaluate the use of human urothelial cells (HUC) seeded on bovine collagen type I-based cell carriers (CCC) in an animal model and to evaluate short-term outcome of the surgical procedure.Methods Four male Gottingen minipigs were used with immunosuppression (cyclosporine A) for this pilot xenograft study. HUC obtained from human benign ureteral tissue were stained by PKH26 and seeded on a collagen cell carrier (CCC). Seven weeks after urethral stricture induction and protective vesicostomy, cell-seeded CCC was implanted in the urethra with HUC luminal and antiluminal, respectively. After two weeks animals were euthanized, urethrography and histological assessment were performed.Results Surgery was technically feasible in all minipigs. Stricture was radiologically established 7 weeks after induction. CCC was visible after two weeks and showed good integration without signs of inflammation or rejection. In the final urethrography, no remaining stricture could be detected. Near porcine urothelium, PKH26-positive areas were found even if partially detached from CCC. Although diminished, immunofluorescence with pankeratin, CK20, E-cadherin and ZO-1 showed intact urothelium in several areas on and nearby CCC.Conclusion Finally, this study demonstrates that the HUC-seeded CCC used as a xenograft in minipigs is technically feasible and shows promising results for further studies.