Background: Efficient cell adhesion and proliferation is a central issue in cell-based tissue engineering, which offers great promise for repair of urethral defects or strictures. This study evaluated the adhesion and growth of rabbit uroepithelium on a surface-modified three-dimensional poly-L-lactic acid (PLLA) scaffold.

Methods: Urethral mucosa were harvested from male New Zealand rabbits and the urothelium were dissociated and then cultured. Immunocytochemistry on cultured uroepithelium for pancytokeratin and uroplakin II and TE-7 confirmed pure populations. After in vitro proliferation, cells were seeded onto a surface-modified urethral scaffold with non-knitted filaments. The morphology and viability of the cells were examined by immunohistochemical and fluorescence staining. Inverted and scanning microscopes were used to document cell growth and adhesion.

Results: Three to five days after primary culture, the uroepithelial cells gradually became confluent, assuming a cobblestone pattern. The filaments of the urethral scaffold had excellent biocompatibility and allowed growth of the uroepithelium, without affecting viability. The uroepithelial cells adhered to and grew well on the scaffold. After 3 - 7 days, the cells grew vigorously and meshes of the scaffold were full of uroepitheliums.

Conclusions: The surface-modified urethral scaffold with non-knitted filaments allows the growth of uroepithelium and can serve as a carrier for the tissue engineering of urethra.

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Article Synopsis
  • - The study compares two decellularization methods (enzyme-detergent and detergent-detergent) to see how they affect the structure and mechanics of human urethral tissue, using samples from 18 individuals.
  • - Both methods successfully removed cellular content while maintaining essential extracellular matrix components like collagen and elastic fibers, indicating they preserved the tissue's structural integrity.
  • - Biomechanical tests showed that the detergent-detergent method resulted in lower stress levels compared to native tissues, and both methods had reduced elasticity, but none were toxic to stem cells, suggesting they are viable for tissue engineering and biobanking.
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Urethroplasty for the management of long-segment urethral strictures associated with lichen sclerosus presents considerable clinical challenges. Oral mucosal grafts are commonly employed but are vulnerable to posttransplantation infection and recurrent stricture formation. Furthermore, the necessity for anesthesia and oral graft harvesting restricts their application in primary healthcare settings.

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Scaffold-based tissue engineering strategies for urethral repair and reconstruction.

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Department of Urology, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai Eastern Institute of Urologic Reconstruction, Shanghai Jiao Tong University, Shanghai 200233, People's Republic of China.

Urethral strictures are common in urology; however, the reconstruction of long urethral strictures remains challenging. There are still unavoidable limitations in the clinical application of grafts for urethral injuries, which has facilitated the advancement of urethral tissue engineering. Tissue-engineered urethral scaffolds that combine cells or bioactive factors with a biomaterial to mimic the native microenvironment of the urethra, offer a promising approach to urethral reconstruction.

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Urethral reconstruction presents a challenging issue in urology, primarily due to the limited availability of alternative materials for repair. The advancement of bioengineering technology has brought new hope to researchers, with a focus on the selection of appropriate biological scaffolds and seed cells. In order to find an ideal alternative material, we used platelet-rich fibrin as the bioscaffold and urothelial cells as the seed cells, meanwhile, we intended to investigate the effect of platelet-rich fibrin on the biological properties of urothelial cells.

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