Investigating the Effect of Ascorbic Acid on the Viability of ‎Mesenchymal ‎Stem Cell Cultured on the Skeletal Muscle ‎Decellularized Scaffold

Tissue engineering is an emerging field based on the three elements of cells, scaffolds, and bioactive molecules, and can be a useful method for treating muscle injuries. The aim of this study is to investigate the effect of ascorbic acid (AA) on the viability of bone marrow mesenchymal stem cells (BM-MSCs) cultured on skeletal muscle decellularization scaffold. First, BM-MSCs were extracted from rat leg bone marrow and cultured in vitro. The identity of the cells was assessed using flow cytometry. The extracted rat skeletal muscle was decellularized using a 1% SDS solution. The decellularization process was investigated by Masson Trichrome, and Alcian blue and DAPI staining.BM-MSCs were cultured on decellularized scaffolds and treated with 1 mM AA for 2 days. Then, the survival and viability of the cells were evaluated using scanning electron microscope (SEM) and MTT methods, respectively.BM-MSCs had a spindle morphology, and the results of flow cytometry showed the expression of CD44 and CD90 and the lack of expression of CD45 and CD34 in more than 90% of the cells. The staining verified the preservation of collagen and glycosaminoglycans and the absence of DNA in the decellularized tissue. MTT results showed that AA significantly increases the viability of BM-MSCs (P<0.05). Also, the SEM results showed that the cells in the group treated with AA were more proliferated. In general, AA can improve the efficiency of muscle tissue engineering by increasing the viability of BM-MSCs.