Thermoresponsive nanofiber composites comprising biopolymers and ZnO nanoparticles with controlled release and antibacterial activity are fascinating scientific research areas. Herein, poly(N-isopropylacrylamide) (PNIPAm) was prepared and mixed with poly(vinyl alcohol) (PVA) in 75/25 and 50/50 weight ratios together with ZnO (0, 1, and 2 phr) to construct nanofiber composites. The morphology of the crosslinked nanofiber composites, ZnO content, and their mechanical behavior were assessed by SEM, EDX, and tensile analyses. The wettability results show an increment in nanofiber surface hydrophobicity by increasing the temperature above the LCST of PNIPAm. The in vitro ZnO release exhibits a faster release profile for the sample with 50 wt% PNIPAm (lower crosslinking density) compared to the one with 25 wt%. Besides, a strong interaction between PVA hydroxyl groups and ZnO can restrict the release content. However, by increasing the temperature from 28 to 32 °C, the relative ZnO release becomes half for both compositions. All crosslinked nanofiber composites demonstrated reliable biocompatibility against L929 fibroblast cells. Agar disc-diffusion and optical density methods showed thermo-controllable antibacterial activity against Staphylococcus aureus upon temperature variation between 28 and 32 °C. Furthermore, in vivo and histological results indicate the potentiality of the prepared multidisciplinary wound dressing for robust wound healing and skin tissue engineering.