A Bovine Collagen Type I-Based Biodegradable Matrix as a Carrier for Tissue-Engineered Urothelium

Martin Vaegler Lisa Daum

doi:10.4172/2157-7633.1000275

Abstract

Engineering of lower urinary tract tissue suitable for reconstructive surgery requires biomaterials as cell carriers, particularly in patients for whom autologous grafts are not available. Matrices should support growth, improve mechanical stability, feature excellent biocompatibility, and fully degrade without signs of scarring at the implantation side. In this study, a new bovine collagen type I-based biodegradable non-cross linked matrix was investigated for its suitability as a carrier for porcine and human urothelial cells in vitro. Initial cell adherence, metabolic activity, and proliferation behaviour of cells isolated from tissue biopsies were analyzed. Constructs were characterized immunohistologically in comparison with matrix-free cell sheets established on plastic surface (=controls). Even for high-density seeding, adherence on collagen cell carrier (CCC) was excellent. Metabolic activity and proliferation of stratifying porcine and human urothelial cells cultured on CCC were comparable to that of controls. Immunofluorescence analysis confirmed epithelial phenotype, cell-cell junction formation and ongoing differentiation of the multilayered urothelium on CCC. This study proved CCC as a suitable carrier for urothelial cells for the future aim for urethral reconstruction.

Highlights

Urogenital impairment resulting from congenital diseases, traumatic injuries, inflammation, tumours, or medical intervention requires reconstructive surgery techniques
For adherence of HUC (Figure 2B), the results for high-density seeding were equivalent between standard plastic surface and collagen cell carrier (CCC) (2.0% vs. 2.2%), whereas adherence was decreased for CCC with lowdensity seeding (2.2% vs. 16.2%)
For Porcine urothelial cells (PUC) cultures on CCC, metabolic activity determined as mean percentage of control values showed a slightly higher and increasing activity from 104% at day 0 to 119% at day 8 after induction of stratification

Summary

Introduction

Urogenital impairment resulting from congenital diseases, traumatic injuries, inflammation, tumours, or medical intervention requires reconstructive surgery techniques. In patients for whom autologous grafts are not available, suitable tissue equivalents are needed for reconstructive purposes [1]. For this reason, tissue engineering (TE) has become a promising technique in recent years [2,3]. The first step in TE techniques for the lower urinary tract include the isolation and the expansion of primary urothelial cells followed by the development of a 3D urothelial differentiated and stratified tissue. Monolayered primary urothelial cell cultures are either induced to differentiate and stratify into matrix-free urothelial tissue or are seeded on biodegradable cell carriers to obtain matrix-stabilized urothelium [5,6,7]

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Conclusion