Background: Despite remarkable development in wound healing management, there is no ideal drug available to address diverse complexity in wound care. This necessitates the development of a comprehensive wound healing formulation. Objectives: The objective of this study was to evaluate the potential of ascorbic acid, castor oil, and gum tragacanth formulation, as an effective wound healing composition. Materials and Methods: A formulation containing ascorbic acid (0.5%), tragacanth gum (6%), and castor oil (25%) is prepared and evaluated for its efficacy on full-thickness mouse excisional wound model. Povidone-iodine was used as a positive control for comparative efficacy. Morphological and histopathological studies were carried out for understanding its wound healing potential. Antibacterial action was tested using disk diffusion method. Safety assessment for oral toxicity and eye and skin irritation was investigated in mouse and rabbit models, respectively. Permeability studies of formulation with excised skin have been evaluated by diffusion chamber, and its absorption in skin and serum was measured with high-performance liquid chromatography method. Results: Wound treated twice a day with formulation showed up to 24% recovery after 24 h and full skin restoration on the 9th day compared to the untreated wound. The morphological and histopathological examinations of wound indicated noteworthy improvement in recovery. Comparative efficacy showed 98% wound recovery in formulation-treated groups compared to 89% with commercially available povidone-iodine ointment after the 9th day. Permeability data clearly indicate enhanced skin uptake in time-dependent manner. The formulation is found to be safe and effective as demonstrated by antibacterial nature, permeability, and supportive safety studies. Conclusion: This study elucidates this formulation possesses strong wound healing potential.