Enhanced Wound Healing, Anti-Inflammatory, and Anti-Oxidative Effects of Vitamin C-Loaded Hyaluronic Acid–Collagen Scaffolds in Preclinical Rat Models

Wound healing is a complex biological process critical for restoring skin integrity after injury. However, chronic wounds present significant clinical challenges due to persistent inflammation, oxidative stress, disrupted collagen synthesis, and susceptibility to infection. Bioactive scaffolds have emerged as promising therapeutic strategies to enhance tissue regeneration by modulating cellular behavior and extracellular matrix (ECM) dynamics. This study explores a hyaluronic acid-collagen (HyCol) scaffold enriched with vitamin C (VC), producing (VC-HyCol) to improve wound healing in preclinical rat models. Hyaluronic acid and collagen, key ECM components, provide structural and biochemical support, while vitamin C acts as both a collagen biosynthesis cofactor and an antioxidant to counteract oxidative stress. Physicochemical characterization confirmed the scaffolds maintained favorable porosity and swelling capacity, while <i>in vitro</i> assessments demonstrated excellent cytocompatibility against human skin fibroblasts and potent antimicrobial activity, particularly against <i>Escherichia coli</i> (<i>E. coli</i>) and <i>Candida albicans</i> (<i>C. albicans</i>). In <i>in vivo</i> studies, the optimal formulation, VC-HyCol-3, significantly accelerated tissue regeneration, achieving the highest wound closure rates by day 14. Mechanistic analysis further confirmed the scaffold's multifaceted therapeutic role; it enhanced antioxidant defenses by upregulating glutathione (GSH) and significantly reducing malondialdehyde (MDA) levels; it suppressed inflammatory mediators (downregulated tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and nuclear factor κB (NF-κB) p65); and it promoted angiogenesis (upregulated vascular endothelial growth factor (VEGF) and CD34) and enhanced collagen deposition. Histological and immunohistochemical evaluations corroborated these findings, showing restored epidermal architecture and minimal inflammatory infiltration in VC-HyCol-3-treated wounds. These findings highlight the potential of this multifunctional scaffold as an advanced wound dressing capable of accelerating healing through structural support, oxidative stress mitigation, and immunomodulation, with significant implications for regenerative medicine.