Development and Evaluation of Indian Mustard Honey-Loaded Fast Dissolving PVA/AG Nanofibers for Enhanced Neuroprotection

Mustard honey (MH) is a rich source of polyphenols, which impart antioxidant, antiapoptotic, and cytoprotective activities in neuronal cells. However, its high-dose requirement for delivering pharmaceutical properties, along with challenges faced by dysphagic individuals, necessitates a controlled delivery system for therapeutic applications. In this study, electrospun nanofibers comprising poly(vinyl alcohol) (PVA) and acacia gum (AG), loaded with varying concentrations of the Indian variety of MH (5–20%), were designed and developed for sublingual administration. The physicochemical properties of PVA/AG/MH nanofibers, including viscosity, conductivity, morphology, and chemical composition, were investigated. Release kinetics analysis showed that MH release followed a non-Fickian mechanism that was best described by the Korsmeyer-Peppas model, and 15% MH-loaded nanofibers achieved an optimal balance between rapid, predictable release and uniform fiber morphology. The antioxidant potential of the nanofibers was assessed using DPPH and ABTS assays, while their neuroprotective effects were evaluated in HT-22 cells subjected to H₂O₂-induced oxidative stress followed by measuring the intracellular reactive oxygen species (ROS), antioxidant gene expression, apoptotic cell population, and mitochondrial membrane potential. PVA/AG nanofibers loaded with 15 and 20% MH concentrations (w/v) showed significantly enhanced antioxidant and neuroprotective activities against H₂O₂-induced oxidative stress by upregulating antioxidant genes including Sod1 (2.6- and 2.3-fold), Sod2 (3.3- and 3.4-fold), Cat (2.9- and 3.1-fold), Gpx (3.1- and 3.3-fold), and Gsta1 (3.9- and 4.5-fold), leading to decreased ROS, reduced apoptotic cell population, and improved mitochondrial membrane potential. In conclusion, PVA/AG/MH nanofibers demonstrated a potent controlled drug delivery system capable of inducing neuroprotective activity via sublingual administration.