Evaluation of Wound Healing Activity (Excision Wound Model) of Ointment Prepared from Ethanolic Leaves Extract and Fractions of Cleistopholis staudtii

Cleistopholis staudtii is a medicinal plant traditionally used in Africa for wound management, but its scientific validation remains limited. This study evaluated the wound-healing efficacy, phytochemical composition, and safety profile of its ethanol extract and solvent fractions. Leaves were macerated in ethanol to obtain a crude extract (19.1% yield), which was fractionated into n-hexane (HF), ethyl acetate (EAF), and n-butanol (BF) fractions, with EAF showing the highest yield (30.4%). Phytochemical analysis revealed abundant flavonoids (8.6%) and phenolics, particularly in EAF and BF. Acute toxicity testing in mice established an LD₅₀ exceeding 5000 mg/kg, indicating high safety. In excision wound models (50 rats), topical application of 10% BF and 10% EAF formulations significantly increased wound closure, achieving near-complete healing by Day 18 (comparable to cicatrin powder, p < 0.05), while petroleum jelly controls showed minimal improvement. LC-MS analysis identified key bioactive compounds, including kaempferol glycosides (antioxidant), eugenol (antimicrobial), and salicin (anti-inflammatory), which likely synergistically promoted tissue regeneration through anti-inflammatory, antimicrobial, and collagen-modulating mechanisms. These findings scientifically validate the traditional use of C. staudtii and highlight its 10% butanol and ethyl acetate fractions as promising candidates for development into standardized wound-healing agents.