Wound dressing is an ancient technique for promoting healing, and modern technology has led to the development of advanced dressings that enhance patient care. Nanofiber-based wound dressings are a medical innovation with enhanced properties, including improved adhesion, reduced infection rates, and increased tissue regeneration. This article focuses on electrospun nanofibrous wound dressing materials produced using the widely adopted method of electrospinning. This article explores several parameters that influence fiber size, including electrical conductivity, electric potential, collector distance, viscosity, flow rate, and surface tension. With Fucoidan (FUC) loading, an increase in the fiber diameter of the control group from 310 nm to 395 nm was observed. This research also examines the use of Halomonas Levan (HL), a polysaccharide, and polyvinyl alcohol (PVA) polymer as wound dressing materials to enhance the mechanical properties of the latter. The incorporation of various concentrations of FUC into PVA-HL electrospun nanofibers yielded diverse effects on tensile strength: an enhancement was observed in the PVA-HL-10FUC formulation, while reductions were noted in the PVA-HL-13FUC and PVA-HL-15FUC formulations. The WST1 assay demonstrated that none of the samples exhibited cytotoxicity up to 72 h, as cell viability increased over time. In conclusion, nanofibrous PVA-HL structures loaded with FUC, which promote tissue regeneration and prevent infection, could be considered a novel wound dressing material.