Twisted stainless steel wires are used as wire electrodes for electrospinning the polyvinyl alcohol (PVA)/zinc citrate nanofiber mats. The morphology and diameter of the nanofibers in PVA/zinc citrate nanofiber mats are evaluated. We measured the antibacterial efficacy against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) of the nanofiber mats. We also examined the cell adhesion affinity of mammalian tissue culture cells on these nanofiber mats. Our results indicate that an increase in zinc citrate increases the viscosity and electrical conductivity of PVA solution. In addition, increasing zinc citrate results in a smaller diameter of nanofibers that reaches below 100 nm. According to the antibacterial test results, increasing zinc citrate enlarges the inhibition zone of S. aureus but only has a bacteriostatic effect against E. coli. Finally, cell adhesion test results indicate that all nanofiber mats have satisfactory cell attachment regardless of the content of zinc citrate.