This study aimed to investigate the histopathological and biochemical effects of Calendula officinalis (C. officinalis) on cutaneous wound healing in rats. Twenty-four Wistar albino rats were were randomly allocated into three groups (n = 8 each): Control, C. asiatica, and C. officinalis. Circular excision wounds (8 mm in diameter) were created in the interscapular region using a biopsy punch under xylazine-ketamine anesthesia. C. asiatica and C. officinalis ointments were applied topically once daily to their respective treatment groups, while glycerin ointment was administered to the Control group. Blood and skin tissue samples were collected on post-injury days 7 and 15 for histopathological and biochemical analyses. Both C. asiatica and C. officinalis groups demonstrated significantly greater wound contraction compared to the Control group. Histopathological evaluation revealed that topical application of C. officinalis exerted anti-inflammatory effects by reducing inflammation and edema, while promoting epithelialization, fibroblast proliferation, and collagen deposition. Biochemical analyses showed significant reduction in malondialdehyde levels, restoration of antioxidant defense system components, and improvement in several biochemical parameters in the C. officinalis group. Topical application of C. officinalis significantly accelerated wound healing by shortening epithelialization time, enhancing connective tissue proliferation, and restoring oxidative balance.