Development of Synergistic Bioactive Hydrogel for Wound Healing Applications

Abstract

This study aims to develop a novel bioactive hydrogel designed to enhance wound healing through the synergistic combination of modified hyaluronic acid (HA), chitosan, and natural bioactive agents such as curcumin, aloe vera, and green tea extracts. The bioactive hydrogel was synthesized utilizing a combination of hyaluronic acid (HA) and chitosan as the foundational structural components. To enhance the therapeutic potential, natural bioactive agents—curcumin (for anti-inflammatory and antioxidant effects), aloe vera extract (for its soothing and regenerative properties), and green tea extract (for its antimicrobial and antioxidant benefits) were incorporated. Chitosan was dissolved in 1% acetic acid solution to a concentration of 2% w/v under continuous stirring until fully solubilized. The modified hyaluronic acid was prepared in phosphate-buffered saline (PBS) at a concentration of 1% w/v, ensuring complete dissolution. Curcumin, aloe vera, and green tea extracts were introduced into the chitosan solution in their respective optimal concentrations to preserve their bioactivity. This step was conducted under controlled conditions to prevent any degradation of the bioactive agents. The prepared HA-modified solution was mixed with the chitosan-bioactive agent mixture in equal volumes. Crosslinking was initiated by the gradual addition of sodium tripolyphosphate (TPP) as a crosslinker, which was added dropwise while maintaining gentle stirring to achieve uniform gelation. The hydrogel formation occurred over a period of 2-3 hours at room temperature, resulting in a stable, biocompatible, and consistent hydrogel matrix. The in-vitro biocompatibility was tested using standard cytotoxicity assays using L929 fibroblast cells, while biodegradability was assessed over time through weight loss measurements in physiological conditions. The hydrogel showed strong antimicrobial activity, toward S. aureus and E. coli, as compared to the control. Histological analysis was also conducted in in-vitro to assess the wound healing ability using the scratch assay test. Overall, the novel bioactive hydrogel, leveraging the synergistic properties of HA-modified hyaluronic acid, chitosan, and natural bioactive agents, presents a promising multifunctional approach to wound healing. Its capacity to enhance hydration, control infection, and mitigate inflammation positions it as a potential next-generation wound care solution.