Potential in two types of collagen scaffolds for urological tissue engineering applications – Are there differences in growth behaviour of juvenile and adult vesical cells?

Abstract

The aging society has a deep impact on patient care in urology. The number of patients in need of partial or whole bladder wall replacement is increasing simultaneously with the number of cancer incidents. Therefore, urological research requires a model of bladder wall replacement in adult and elderly people. Two types of porcine collagen I/III scaffolds were used in vitro for comparison of cell growth of two different pig breeds at different growth stages. Scaffolds were characterised with scanning electron and laser scanning microscopy. Urothelial and detrusor smooth muscle cells were isolated from 15 adult Göttingen minipigs and 15 juvenile German Landrace pigs. Growth behaviour was examined in cell culture and seeded onto the collagen scaffolds via immunohistochemistry, two-photon laser scanning microscopy and a viability assay. The collagen scaffolds showed different structured surfaces which are appropriate for seeding of the two different cell types. Moisturisation of the scaffolds resulted in a change of the structure. Cell growth of German Landrace urothelial cells and smooth muscle cells was significantly higher than cell growth of the Göttingen minipig cells. Seeding of scaffolds with both cell types from both pig races was possible which could be shown by immunohistochemistry and two-photon laser scanning microscopy. Growth behaviour on the scaffolds was significantly increased for the German Landrace compared to Göttingen minipig. Nevertheless, seeding with the adult Göttingen minipig cells resulted in a closed layer on the surface and urothelial cells and smooth muscle cells showed increasing growth until day 14. The results show that these collagen scaffolds are adequate for the seeding with vesical cells. Moreover, they seem appropriate for the use as an in vitro model for the adult or elderly as the cells of the adult Göttingen minipig too, show good growth behaviour.

Two differentially structured collagen scaffolds for potential urinary bladder augmentation: proof of concept study in a Göttingen minipig model

“…A future approach should entail a partial cystectomy and therefore, the fabrication of a larger prototype is definitely necessary. Also, as we were already proposing in our in vitro study [37], a combination of both scaffolds could be advantageous and provide a more convenient support for both cell types in vitro and in vivo and lead to a more stable implantation site.…”

“…Autologous UCs and SMCs were isolated as described previously [37]. Briefly, the urothelium and detrusor muscle were dissected from the bladder biopsy and the cells mechanically dissociated.…”

“…Immunohistochemistry with pancytokeratin (PanCK) and α-smooth muscle actin (α-SMA) was performed as described previously using the antibodies mentioned in Table 1 [37]. Briefly, sections were deparaffinized in descending ethanol solutions.…”

Future Research Directions in the Design of Versatile Extracellular Matrix in Tissue Engineering

“…The growth rates of German Landrace urothelial cells and smooth muscle cells were significantly higher on collagen-composed scaffolds. This growth behavior showed that these collagen scaffolds were adequate for urologic tissue regeneration [ 64 , 65 ].…”

The Impact of γ-Irradiation and EtO Degassing on Tissue Remodeling of Collagen-based Hybrid Tubular Templates

“…Second, young (porcine) animals were used, known for their fast growth, specific skin properties, and regeneration. 21 This may also cause excessive wound contraction. Third, the choice of sterilization technique may have extensively influenced the final outcome since γ-sterilization enhances Vicryl biodegradability from 2 months to less than 1 month.…”