In this paper, hydroalcoholic solutions of Thymus vulgaris, Salvia officinalis folium, and Hyperici herba were used in combination with poly (vinyl alcohol) with the aim of developing novel poly (vinyl alcohol)-based nanofiber mats loaded with phytotherapeutic agents via the electrospinning technique. The chemical structure and morphology of the polymeric nanofibers were investigated using Fourier Transform Infrared Spectroscopy (FTIR) and Scanning Electron Microscopy (SEM). The addition of Thymus vulgaris, Salvia officinalis folium, and Hyperici herba extracts to the pure polyvinyl alcohol fibers led to changes in the morphology of the fibers and a reduction in the fibers’ diameter, from 0.1798 µm in the case of pure polyvinyl alcohol to 0.1672, 0.1425, and 0.1369 µm in the case of polyvinyl alcohol loaded with Thymus vulgaris, Salvia officinalis folium, and Hyperici herba, respectively. The adapted Folin–Ciocalteu (FC) method, which was used to determine the total phenolic contents, revealed that the samples of PVA–Hyperici herba and PVA–Thymus vulgaris had the highest phenol contents, at 13.25 μgGAE/mL and 12.66 μgGAE/mL, respectively. Dynamic water vapor measurements were used in order to investigate the moisture sorption and desorption behavior of the developed electrospun materials. The antimicrobial behavior of these products was also evaluated. Disk diffusion assay studies with Escherichia coli, Staphylococcus aureus, and Methicillin-resistant Staphylococcus aureus were conducted on the developed nanofibers in order to quantify their phytotherapeutic potential.