Enhancing Wound Regeneration Potential of Fibroblasts Using Ascorbic Acid-Loaded Decellularized Baby Spinach Leaves

Abstract

Decellularization of plant tissues is an emerging route to fabricate scaffolds for tissue engineering and regenerative medicine. Although significant progress has been made in the field of plant tissue decellularization, functionalization of plant scaffolds is still an emerging field, and loading them with L-ascorbic acid to promote skin regeneration has not yet been reported. L-ascorbic acid is an antioxidant that plays a key role in collagen synthesis as a cofactor of lysyl hydroxylase and prolyl hydroxylase. It has been shown to have significant importance in physiological wound healing by stimulating fibroblasts to produce collagen at both the molecular and the genetic levels. In this work, we aimed to fabricate an ascorbic acid-releasing bioactive scaffold by introducing a stable form of ascorbic acid, L-ascorbic acid 2-phosphate (AA2P), into decellularized baby spinach leaves and investigated its biological activity in vitro. Our results demonstrated that AA2P could be easily introduced into decellularized baby spinach leaf scaffolds and subsequently released within the effective dose range. AA2P-releasing baby spinach leaves were found to increase metabolic activity and enhance collagen synthesis in L929 fibroblasts after 21 days. In conclusion, this study demonstrated the fabrication of a novel functionalized skin tissue engineering scaffold and made a significant contribution to the fields of plant decellularization and skin tissue engineering.