Electrospun Nanofibers Loaded with Verbena officinalis Extract as Multifunctional Bioactive Wound Dressings.

Background/Objectives: Verbena officinalis has long been used in traditional medicine for treating skin disorders, yet its potential in advanced wound-dressing systems remains insufficiently explored. This study aimed to characterize V. officinalis extracts obtained using different solvents, evaluate their antimicrobial, antibiofilm, cytotoxic, and wound-healing activities, and develop electrospun polycaprolactone-polyethylene glycol (PCL-PEG) nanofibers as a delivery platform for the most active extract. Methods: Extracts were chemically characterized by LC-MS and evaluated against clinical bacterial, yeast, and dermatophyte isolates associated with skin infections. Antibiofilm activity was assessed against Staphylococcus lugdunensis. The most active hydroalcoholic extract (EW7.5) was incorporated into PCL-PEG nanofibers (1-10%, w/w). Metabolite release was monitored by semi-quantitative LC-MS analysis. Cytotoxicity and wound-healing activity were evaluated using HaCaT keratinocytes, while in vivo efficacy was assessed in a full-thickness excisional wound model in Dark Agouti rats (five animals per group in two independent experiments). Results: EW7.5 exhibited the strongest antimicrobial activity, with MIC values of 0.06 mg/mL against all tested bacterial and Candida strains and 0.06-0.5 mg/mL against dermatophytes. It inhibited S. lugdunensis biofilm formation by 79.43% at MIC/4 and reduced pre-formed biofilm biomass by 64.98% at 2 × MBC. EW7.5 and EW7.5-loaded nanofibers showed no cytotoxicity up to 400 μg/mL. In the scratch assay, the free extract achieved 42.34 ± 1.58% wound closure, while nanofibers containing 10% EW7.5 achieved 25.87 ± 2.52%, compared to 7.73 ± 0.07% for the control. In vivo, EW7.5-loaded membranes significantly accelerated wound closure and the 10% formulation significantly improved extracellular matrix deposition and vascular remodeling compared to the control membrane. Conclusions: EW7.5-loaded PCL-PEG nanofibers combine antimicrobial, antibiofilm, biocompatible, and wound-healing properties, supporting their potential as multifunctional wound dressings. Nevertheless, further studies with mechanistic investigations are required to fully establish their therapeutic applicability.