Study of the wound healing potential of bone marrow stromal cell-seeded biocompatible chitosan-alginate scaffold containing biological materials of alpha-tocopherol and ascorbic acid in hypoxia conditions

Abstract

Introduction: Hydrogel scaffolds are commonly used for skin tissue engineering. Studies have shown that chitosan, alginate, ascorbic acid and α-Tocopherol are highly effective in promoting wound healing. Ascorbic acid and α-Tocopherol, have exhibited a notable capacity in advancing wound healing by increasing reparative mediators. Additionally, subjecting of MSCs under hypoxic conditions enhances their therapeutic capabilities. Our study aims to investigate the repetitive potential of a chitosan/alginate composite hydrogel loaded with Ascorbic acid/α-Tocopherol incorporating hypoxic-MSCs as a promising substitute in wound healing methodologies. Methods: Chitosan/Alginate was blended with Ascorbic acid and α-Tocopherol. The physicochemical properties such as swelling and biodegradation rate, wettability and FTIR were analyzed. MSCs were seeded on the scaffold surface and survival rate analyzed using MTT. Scaffold patches were transplanted on rat model of experimentally induced cutaneous injuries. After 7, 14 and 21 days, post-transplantation, rats were euthanized and cutaneous samples were subjected to Hematoxylin-Eosin staining. Results: The results demonstrated that the prepared composite hydrogels were highly porous, with interconnected pores. Based on wettability and hemolysis tests, these scaffolds were hydrophilic and compatible with blood. MTT assay, DAPI, and H&E staining also showed that the prepared hydrogels provide a suitable substrate for cell growth and proliferation. Finally, based on real-time PCR, increased expression levels of VEGF and TGF-β1 were observed in hypoxic-MSCs cultured on the prepared hydrogels. The in vivo study results indicate that the prepared composite hydrogel, had better wound closure, and the highest wound healing was observed in the H-CA/AA/TP group. Conclusion: In conclusion, this study suggests that Chit/Alg composite hydrogels, supplemented with AA and TP and seeded with hypoxic-MSCs, can be effectively used in the treatment of skin injuries in clinical trials.