The most important properties of performant wound dressings are biocompatibility, the ability to retain large amount of exudate and to avoid complications related with persistent infection which could lead to delayed wound healing. This research aimed to obtain and characterize a new type of antimicrobial dressings, based on zinc oxide/sodium alginate/polyvinyl alcohol (PVA). Zinc oxide nanostructures, obtained with different morphology and grain size by hydrothermal and polyol methods, are used as antimicrobial agents along with sodium alginate, which is used to improve the biocompatibility of the dressing. The nanofiber dressing was obtained through the electrospinning method. Characterization techniques such as X-ray diffraction (XRD) and scanning electron microscopy (SEM) were performed to determine the structural and morphological properties of the obtained powders and composite fibers. Their antimicrobial activity was tested against Gram negative Escherichia coli (E. coli), Gram positive Staphylococcus aureus (S. aureus) bacteria and Candida albicans (C. albicans) yeast strains. The in vitro biocompatibility of the obtained composites was tested on human diploid cells. The obtained results suggest that the composite fibers based on zinc oxide and alginate are suitable for antimicrobial protection, are not toxic and may be useful for skin tissue regeneration if applied as a dressing.