Collagen cell carriers seeded with human urothelial cells for urethral reconstructive surgery: first results in a xenograft minipig model

World J Urol. 2017 Jul;35(7):1125-1132. doi: 10.1007/s00345-016-1959-3. Epub 2016 Oct 25.

Abstract

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 Göttingen 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.

Keywords: Collagen matrix; Human urothelial cells; Reconstructive urology; Tissue engineering; Urethra; Xenograft.

MeSH terms

  • Animals
  • Cattle
  • Cell Transplantation / methods*
  • Collagen Type I / physiology
  • Disease Models, Animal
  • Heterografts
  • Humans
  • Male
  • Models, Anatomic
  • Reconstructive Surgical Procedures / methods*
  • Swine
  • Swine, Miniature
  • Tissue Engineering / methods*
  • Treatment Outcome
  • Urethral Stricture / surgery*
  • Urologic Surgical Procedures, Male / methods*
  • Urothelium / cytology*

Substances

  • Collagen Type I