Background Henna (Lawsonia inermis) is a medicinal plant with significant cultural and traditional value. Its leaves have been used for centuries in various cosmetic and medicinal applications. However, the phytochemical composition and wound healing potential of henna from different regions have not been thoroughly investigated. Material and Methods Samples of Lawsonia inermis leaves were collected from three locations in Uttar Pradesh: Sharavasti, Bahraich, and Lucknow. Phytochemical analysis was performed on water-soluble extracts to determine the concentrations of phenolic compounds, flavonoids, tannins, and saponin. Swiss albino mice were used to evaluate the efficacy of specific compositions derived from these extracts in injury rehabilitation. Chromatographic analysis on Sephadex-G50 gel was used to determine the lawsone content in the extracts. Results Phytochemical analysis revealed significant concentrations of phenolic compounds, flavonoids, and tannins in the water-soluble extracts. Sharavasti henna extract exhibited higher levels of phenolic compounds (13.48%), flavonoids (9.25%), and tannins (2.57%), while Bahraich henna extract had a greater saponin concentration (0.32%). Chromatographic analysis confirmed higher lawsone content in Sharavasti henna. Application of 10% henna extract in petroleum jelly significantly reduced burnt area with contraction percentage (CP) and complete re-epithelialization duration (CRD) similar to 1% Flamazine treatment. Conclusion This study suggests the potential of henna-based formulations rich in phenolic chemicals for dermatological, cosmetic, and medicinal applications. The findings highlight the importance of phytochemical analysis and wound healing potential of henna from different regions, which can be used to develop effective henna-based products for various applications.