BackgroundTraditional medicine in form of decoctions has been known for ages to possess wound healing abilities. One such traditional formulation mentioned in Indian literature Charak Samhita Chikitsa Sthanam isKampillakadi Tailaand tremendous information is available on its implication in the treatment of skin cuts and wounds, diseases, or bacterial infections. This research paper focuses on studying the wound healing property of one such herbal proprietary formulation known as a wound healing oil, derived fromKampillakadi Tailafortified with root extract ofWagatea spicata(VIKHPF).ObjectiveThe current research is aimed at studying chemical profiling, antioxidant activity, antimicrobial efficacy,in-vitrocell proliferating, andin-vitrowound healing activity of this VKHPF.Materials and methodsThe chemical characterization of VKHPF was done by gas chromatography- fatty acid methyl esters GC-FAME analysis for lipid analysis and gas chromatography high-resolution mass spectrometry (GC-HRMS)for revealing its chemical constituents. Proliferation and migration are two underlying mechanisms involved in the healing of wounds. Hence,in-vitrostudies such as cell proliferation assay andin-vitroscratch test on NIH/3T3 mice fibroblast cell line were conducted were used to determinein-vitrowound healing capacity of VKHPF. The oil was also tested for antioxidant effect (DPPH assay) and anti-microbial potential (Time kill test).ResultsThe GC-HRMS and GC-FAME analyses revealed rich medicinally important fatty acids and vitamins were present in VKHPF, such as oleic acid, hexadecanoic acid, squalene, α, γ-tocopherol, γ-sitosterol, and benzoic acid. VKHPF at 0.5 mg/ml in media without serum showed 164.00 ± 0.011% cell viability with 64.00% cell proliferation in contrast to media containing serum (100% cell viability). At the same concentration, the wound closure was 98% for VKHPF. The oil sample possessed antioxidant activity with an IC50value of 3.5 mg/ml and antimicrobial activity againstStaphylococcus aureusandPseudomonas aeruginosawhen tested using Time Kill Activity.ConclusionThis study is the first to report the use of Vakeri fortifiedKampillakadi Tailaherbal proprietary formulation (VKHPF) inin-vitrowound healing and the present data suggest that it can form a part of modern medicine.