In Vitro and In Vivo Wound Healing Activity of Dhatakyadi Yogam

Abstract

Dhatakyadi Yogam (DK) was formulated as an ointment using hydroalcoholic extracts of Dhataki (Woodfordia fruticosa (L.) Kurz) flowers and Lodhra (Symplocos racemosa Roxb.) stem bark with ghee and beeswax. Materials and Methods: Hydroalcoholic extracts of Dhataki flowers and Lodhra stem bark were formulated into ointments at 20% w/w (DKO1) and 10% w/w (DKO2). In-vitro wound-healing activity was evaluated using MTT assay (5–15µg/mL), scratch assay, and chick chorioallantoic membrane assay. In-vivo wound-healing activity was evaluated using an excision wound model in Wistar rats divided into five groups: GI (untreated control), GII (povidone iodine), GIII (DKO1), GIV (DKO2), and GV (ointment base). Wound contraction, epithelialization time, and histopathology were assessed. Data were analyzed using one-way ANOVA, with p<0.05 considered statistically significant. Results: Pharmacognostical and physicochemical parameters were in accordance with API standards, and phytochemical analysis confirmed the presence of phenols, steroids, flavonoids, and alkaloids. Maximum cell viability was observed at 10µL (Dhataki), 5µL (Lodhra), and 5µL (DK). DK showed the highest wound closure in the scratch assay at 24 h (98.87±0.85%). Peak angiogenesis was observed in the CAM assay at 5µL (Dhataki), 10µL (Lodhra), and 5µL (DK). In the excision wound model, DKO2 exhibited maximum wound contraction on day 14 (96.94±0.94%) and the shortest epithelialization period (10.4±0.55 days). Conclusion: DKO2 demonstrated significant wound-healing efficacy by enhancing fibroblast activity, angiogenesis, wound contraction, and epithelialization, supporting its potential for further clinical evaluation.