Phytochemical Profiling and Multi-Target Wound-Healing Potential of Millettia extensa (Benth.) Baker Stem Bark...

Millettia species (Fabaceae) are traditionally used across parts of Asia and Africa for managing wounds, skin infections, and inflammatory disorders. Millettia extensa (Benth.) Baker is traditionally associated with the management of wounds and inflammatory conditions. However, its stem bark has not been systematically evaluated for wound-healing activity.

To investigate the wound-healing potential of methanol (MEM) and aqueous (MEA) extracts of M. extensa stem bark using integrated in-silico, in-vitro and in-vivo experimental approaches.

MEM and MEA were subjected to preliminary phytochemical screening, estimation of total phenolic and flavonoid contents, and UHPLC-ESI-QTOF-MS/MS profiling. The shortlisted phytoconstituents were subsequently analyzed using network pharmacology and molecular docking against AKT1, IL-1β, STAT3 and TNF-α, while isoliquiritigenin was further evaluated by 200-ns molecular dynamics simulation. The extracts were further evaluated for antioxidant activity using the DPPH assay, antimicrobial activity using agar well diffusion and broth microdilution assays, acute toxicity and wound-healing activity using the HaCaT scratch-wound assay, carrageenan-induced paw edema model, and excision and incision wound models. Extract-based ointments were evaluated in the topical wound-healing models.

MEM contained higher phenolic (108.59 mg GAE/g) and flavonoid (71.84 mg QE/g) contents than MEA (49.28 mg GAE/g and 40.59 mg QE/g, respectively), with 28 and 63 tentatively annotated compounds, respectively. Network pharmacology identified 86 shared wound-healing targets, among which AKT1, TNF, STAT3, and IL-1β emerged as key hub targets. Isoliquiritigenin exhibited consistent multi-target docking affinities (-9.1 to -6.8 kcal/mol) and the isoliquiritigenin-AKT1 complex showed relatively stable conformational behaviour during the 200-ns molecular dynamics simulation. MEM showed stronger DPPH radical-scavenging activity (IC₅₀ = 64.01 μg/mL) and greater antimicrobial activity, with a MIC of 1.56 mg/mL against P. aeruginosa, S. aureus, and S. epidermidis, compared with MEA. Both extracts showed no observable signs of acute toxicity under the tested conditions. Notably, the 10 % MEM ointment produced 99.2 ± 1.1% excision wound closure, 42.40 ± 1.83 μg/mg hydroxyproline content, and tensile strength comparable to the standard treatment.

Collectively, the present findings indicate that Millettia extensa stem bark, particularly the methanol extract, has promising wound-healing potential, with the observed activity accompanied by antioxidant, antimicrobial, anti-inflammatory, and wound-repair effects. The integrated experimental and computational findings provide a basis for further mechanistic studies and validation of its potential application in topical wound management.