Wound-Healing Potential of a Novel Helichrysum italicum Film-Forming Gel: Enhanced Bioactivity Through Sustained Delivery and Fibroblast Modulation

Wound healing is a highly orchestrated process that can be impaired under pathological conditions, highlighting the need for advanced topical formulations. This study aimed to comparatively evaluate Helichrysum italicum essential oil (EO) and a novel film-forming gel (FFG) incorporating this EO in an in vitro fibroblast-based wound model. Formulations were prepared using polyvinyl alcohol, carboxymethyl cellulose, glycerol, polysorbate 80, and ethanol, yielding stable and skin-compatible gels. Chemical analyses (GC–MS and total phenolic content) verified the presence of bioactive sesquiterpenes and phenolic compounds, while physical characterization confirmed appropriate pH, viscosity, and homogeneity. Biological assays revealed a biphasic, dose-dependent effect of EO: low-to-intermediate concentrations (250–500 µg/mL) enhanced fibroblast viability and proliferation, whereas a high concentration (2 mg/mL) induced cytotoxicity. Additionally, antioxidant, molecular (immunocytochemistry, qRT-PCR, Western blotting), and collagenase inhibition assays were conducted. Among the gel formulations, F2 (0.5% w/w EO) exhibited the most favorable profile, significantly improving cell viability, proliferation, and migration compared to EO alone. Antioxidant analyses showed that F2 markedly reduced intracellular superoxide production and lipid peroxidation while increasing glutathione levels. Molecular evaluations demonstrated significant upregulation of extracellular matrix-related markers (COL1A1, FN1, and MMP7), indicating coordinated ECM synthesis and remodeling. F2 also inhibited collagenase activity, supporting the preservation of structural proteins. Incorporation of H. italicum EO into a film-forming gel enhanced bioactivity, likely via improved local retention and a protective milieu. Although confined to an in vitro model and single polymer, these findings warrant further preclinical investigation of EO-loaded FFGs in wound healing.