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Urethroplasty performed with an autologous urothelium-vegetated collagen fleece to treat urethral stricture in the minipig model

  • Karl-Dietrich SievertEmail author
  • L. Daum
  • S. Maurer
  • P. Toomey
  • M. Vaegler
  • S. Aufderklamm
  • B. Amend
Topic Paper
  • 11 Downloads

Abstract

Introduction and objective

Tissue-engineered materials in urethral reconstructive surgeries are a promising field for innovative therapy. Collagen matrices increase stability of cell-based implants and can promote viability and proliferation of urothelial cells. In this study, a collagen type I-based cell carrier (CCC) with stratified multi-layer autologous urothelium was used for urethroplasty after induction of urethral stricture in eight minipigs.

Materials and methods

Minipigs underwent surgical procedures to induce urethral stricture by thermocoagulation. Simultaneously, bladder tissue was harvested. Urothelial cells were expanded, labeled with PKH26 and seeded onto CCC in high density. 3 weeks after strictures were induced and verified by urethrography, minipigs underwent urethroplasty using the seeded CCC. Two animals were euthanized after 1, 2, 4, and 24 weeks. Urethras were histologically examined for integration and survival of seeded CCC. In vivo phenotype of multi-layered urothelium matrix constructs was characterized via immunofluorescence staining with pancytokeratin, CK20, p63, E-cadherin and ZO-1.

Results

Seeded CCCs showed excellent stability and suturability after manipulation and application. Transplanted cells were detected using positive PKH26 fluorescence up to 6 months after labeling. Urothelium matrix implants integrated well into the host tissue without sign of inflammation. Animals showed no sign of rejection or stricture recurrence (urethrography) at any time during experimental period. Immunofluorescence analysis confirmed epithelial phenotype, junction formation and differentiation after 2 weeks.

Conclusion

CCC can be suitable for urologic reconstructive surgeries and represents a promising option for clinical application. Longer follow-up results are required to exclude re-occurrence of stricture reformation.

Keywords

Urethroplasty Urethral stricture Urothelium Biomaterial Collagen matrix Regenerative medicine Large animal model Cell collagen carrier Urothelium Reconstructive therapy 

Notes

Acknowledgements

The study was supported by Viscofan BioEngineering Germany (CU2/12). The authors would also like to thank Kathyrn Emily Nelson of KNM Communications for her editorial assistance.

Author contributions

KDS: project development, supervision, analysis and supervision of data, manuscript writing, critical revision of manuscript; LD: data collection and manuscript drafting; SM: data collection; PT: manuscript writing/editing; MV: data collection/management, analysis and supervision of data, manuscript drafting; SA: data collection/management, analysis and supervision of data, critical revision of manuscript; BA: analysis and supervision of data, analysis and supervision of data, critical revision of manuscript.

Compliance with ethical standards

Conflict of interest

The author declare that there no conflicts of interests.

Statement of welfare of animals

All applicable, international, national and/or institutional guidelines for the care and use of animals were followed.

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Laboratory of Tissue Engineering, Department of UrologyEberhard-Karls-UniversityTübingenGermany
  2. 2.Department of UrologyEberhard-Karls-UniversityTübingenGermany
  3. 3.Department of UrologyKlinikum LippeDetmoldGermany
  4. 4.Department of UrologyUniversity Clinic of ViennaViennaAustria

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