Synthesis of metal oxide nanoparticles-polymer nanocomposites is an emerging strategy in nanotechnology to improve targeted delivery and reduce the toxicity of nanoparticles. In this study, we report biological effects of previously described hybrid nanocomposites containing dextran-graft-polyacrylamide/zinc oxide nanoparticles (D-PAA/ZnO NPs) prepared from zinc sulfate (D-PAA/ZnONPs(SO42-)) and zinc acetate (D-PAA/ZnONPs(-OAc)) focusing primarily on their antimicrobial activity. D-PAA/ZnONPs(SO42-) and D-PAA/ZnONPs(-OAc) nanosystems were tested in a complex way to assess their antioxidant activity (DPPH assay), antidiabetic potential (α-amylase inhibition), DNA cleavage activity, antimicrobial, and antibiofilm activity. In addition, the toxicity of D-PAA/ZnONPs(SO42-) and D-PAA/ZnONPs(-OAc) nanosystems against primary murine splenocytes was tested using MTT assay. The studied nanosystems inhibited E.coli growth. For all the investigated strains, minimum inhibitory concentrations (MICs) of D-PAA/ZnONPs(SO42-) and D-PAA/ZnONPs(-OAc) were in the range of 8mg/L-128mg/L and 16mg/L-128mg/L, respectively. The nanocomposites demonstrated effective antibiofilm properties as 94.27% and 86.43%. The compounds showed good antioxidant, anti-α-amylase, and DNA cleavage activities. D-PAA/ZnONPs(SO42-) and D-PAA/ZnONPs(-OAc) nanosystems reduced cell viability and promoted cell death of primary murine spleen cells at concentrations higher than those that proved to be antibacterial indicating the presence of therapeutic window. D-PAA/ZnONPs(SO42-) and D-PAA/ZnONPs(-OAc) nanosystems show antioxidant, antidiabetic, DNA cleavage, antimicrobial, and antibiofilm activity against the background of good biocompatibility suggesting the presence of therapeutic potential, which should be further investigated in vivo.