Preparing Sheep Bladder Scaffold and Examining the Differentiation of Mesenchymal Stem Cell into Myocytes on Scaffolds

Background: Tissue engineering may be used to repair, preserve, or improve tissues and organs. In this regard, acellular biological scaffolds are mainly used to reconstruct damaged tissues in regenerative medicine. Objectives: The present study examined the in vitro process of myocytes differentiated from bone marrow mesenchymal stem cells (BM‐MSCs) on the sheep bladder scaffold induced by 5-azacytidine. Methods: Decellularization was performed using a mixed method (physical and chemical) to prepare scaffolds kept at -20°C. The 5-azacytidine was used to induce BM‐MSCs to myocytes. Moreover, the muscle-specific gene expression (Desmin, α-Actinin, Myo D) was evaluated using the RT-PCR method. Results: It was revealed that BM‐MSCs on the scaffold had high proliferation and differentiation potentials. Desmin and α-Actinin gene expression marked the differentiation at the end of the fourth week. Moreover, the results of Masson’s trichrome staining at the end of the second, third and, fourth weeks also indicated that the first differentiation signs emerged at the end of the second week. Furthermore, differentiation reached its maximum level during the fourth week. Conclusions: According to the findings, combining physical and chemical methods was the best technique to prepare the bladder scaffold so that the bone marrow mesenchymal stem cells can be differentiated into myocytes on the bladder scaffold affected by 5-azacytidine (5 µmol), and As the induction time increases to day 28, myocyte cells become more developed.