In Vitro Evaluation of Acellular Skeletal Muscle-De-rived Hydrogel Containing ۵-Azacytidine on The Behavior of Muscle-Derived Stem Cells

Objective: Finding new and appropriate biomaterials/scaffolds is one of the most important aspects in skeletal muscle tissue engineering. Among the biomaterials, decellularized skeletal muscle-derived biomaterials (e.g. hydrogels and scaffolds) are a promising strategy for improving muscle tissue engineering. Moreover, regulatory factors such as ۵-azacytidine (a DNA meythltransferase inhibitor) can induce myogenesis in different type of stem cells, as well as it can regulate viability, migration and secretion. Here, we studied the effect of ۵-Aza incorporated muscle-derived hydrogel in muscle regeneration in vitro.Materials and Methods: Wistar rat skeletal muscles were de-cellularized by sodium dodecyl sulphate (SDS ۱%). DNA con-tent and histological analysis was carried out on acellular and native muscle tissues. Then, the muscle extra cellular matrix (ECM) was enzymatically digested using pepsin and then mus-cle-derived hydrogel was prepared. ۵-azacytidine (۵ µm) was incorporated into the hydrogel and muscle-derived stem cells (MDSCs) were cultured within the hydrogels. The viability of MDSCs was analyzed using Live/Dead staining.Results: DNA content analysis, Masson's trichrome (MT), He-matoxylin and eosin (HandE) staining and scanning electron microscope (SEM) confirmed the cell elimination and mainte-nance of the ECM structure after decellularization. Live/Dead assay showed that hydrogel incorporated with ۵-Aza supports MDSCs viability in vitro.Conclusion: Skeletal muscle-derived hydrogel along with ۵-azacytidine supports MDSCs viability. In addition, the use of myogenic chemical such as ۵-azacytidine and muscle-derived biomaterials might be a promising and useful approach for clin-ical applications.

کلیدواژه ها:

۵-Azacytidine ، Muscle-Derived Stem Cells ، Hydro-gel ، Acellular Skeletal Muscle