Acellular human glans extracellular matrix as a scaffold for tissue engineering: in vitro cell support and biocompatibility

ABSTRACT Objectives: Diseases of the genitourinary tract can lead to significant damage. Current reconstructive techniques are limited by tissue availability and compatibility. This study aims to assess if the decellularized human glans can be used as a biomaterial for penile reconstruction. Materials and Methods: Samples of the glans matrices were descellularized. We evaluate the presence of collagen type I and III, and elastic fibers. Biocompatibility assays were performed to assess the cytotoxic and non-cytotoxic interactions between the acellular matrix and 3T3 cells. The matrices were seeded with mesenchymal stem cells and were assessed for viability and integration of these cells. Biomechanical tests in native tissue, descellularized matrix and seeded matrix were performed to characterize their biomechanical properties. Results: The tissue architecture of the decellularized matrix of human glans was preserved as well as the maintenance of the biomechanical and biological properties. The analyzes of glans seeded with mesenchymal stem cells revealed the integration of these cells to the matrices, and its viability during two weeks "in vitro". Conclusion: The decellularization process did not alter the biological and biomechanical characteristics of the human glans. When these matrices were seeded they were able to maintain the cells integrity and vitality.

Penile reconstruction using mesenchymal stem cells

PURPOSE: To compare the reconstruction of corpus cavernosum segments when seeded with mesenchymal stem cells and when stem cells are infused intravenously.METHODS: Sixteen New Zealand rabbits were submitted to reconstruction of the corpus cavernosum and distributed in Group A - decellularized matrices, Group B - decellularized matrices seeded with mesenchymal stem cells Group C - decellularized matrices submitted to intravenous infusion of mesenchymal stem cells. The mesenchymal stem cells were obtained by bone marrow aspiration. The venous filling aspect of the distal end of the corpus cavernosum was evaluated and the specimens were submitted to histological analisis and to immunohistochemistry. Cavernosometry was done in one animal of each groupRESULTS: Three animals on B and three animals on C presented full filling of distal end of the corpus cavernosum. No animals in A presented filling of the distal end of corpus cavernosum. At cavernosometry the animal on B attained 50 cmH2O, on C 110 cmH2O and on A 20 cmH2O. Trabeculae forming cavernous sinuses were found in groups B and C.CONCLUSION: The reconstruction of corpus cavernosum using descellularized matrices and mesenchymal stem cells, either by intravenous injection or directly seeded is possible, with growth of corpus cavernosum-like tissue.

Integration of collagen matrices into the urethra when implanted as onlay graft

Objective To assess the integration of decellularized heterologous collagen matrices into the urethra, when implanted with no cells or when seeded with autologous smooth muscle cells. Materials and Methods Eighteen New Zealand rabbits were randomly assigned to two groups: Group I (n = 9) - animals undergoing urethral segment resection with interposition of a patch of heterologous collagen matrix seeded with autologous smooth muscle cells; Group II (n = 9) - animals undergoing resection of a urethral segment with interposition of a decellularized heterologous collagen matrix patch. Two animals from each group were sacrificed on postoperative days seven, fourteen and twenty-eight; three animals from each group were sacrificed at the end of three postoperative months. At the end of the third month one animal from each group underwent urethroscopy for urethral integrity assessment and one animal from each group had its microcirculation image captured by a SDF device (Side-stream Dark Field - Microscan Analysis Software). One animal from each group in each euthanasia period underwent cystourethrography so as the urethra could be viewed at flow time. The matrices integration was assessed through histological examination using hematoxylin and eosin (H&E), Masson trichrome (MT), Picrosirius red and Von Willebrand staining. In a blind study with two pathologists all the slides were studied. Results The matrices whether seeded or not with autologous muscle cells were able to restore the architecture of the urethra, but were eliminated from the first week on, before incorporation. Microcirculation of the neourethra, at the end of the third month, showed the same characteristics as a normal urethra in both groups of animals. Conclusion Natural heterologous matrices implanted in the urethra as onlay graft were not incorporated into its walls but were able to fully restore the ...