The tricomponent composite chitosan/polyvinyl pyrrolidone (CS/PVP) with different weight ratios of (0, 20, 40, 60, and 80 wt%) of copper–hydroxyapatite (Cu-HAP) were prepared by solvent casting technique, which was characterized by X-ray diffraction, Fourier-transform infrared, and scanning electron microscopy with energy-dispersive X-ray to confirm the formation of Cu-HAP/CS/PVP composite. The Cu-HAP/CS/PVP composite with 80 wt% Cu-HAP showed 98.73 ± 1.14% of porosity with highest tensile strength (101.45 ± 0.98 MPa) and less swelling percentage (19.51 ± 1.03%) compared to others. In vitro antimicrobial activity was investigated against bacteria (Staphylococcus aureus, Bacillus subtilis, and Escherichia coli) and fungus (Candida albicans, Penicillium notatum, and Rhizopus stolonifer). In vitro hemocompatibility study proves that the Cu-HAP/CS/PVP composites are blood compatible with the hemolytic ratio of less than 2%. In vitro bioactivity study revealed the formation of apatite on the optimized Cu-HAP/CS/PVP composite (80 wt% of Cu-HAP) in simulated body fluid (SBF) solution. ICP-OES (Inductively Coupled Plasma-Optical Emission Spectroscopy) analysis was used to find out the leaching of Ca, P, and Cu ions from the SBF. In vitro biocompatibility was studied against human osteosarcoma cell line by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-DiphenyltetrazoliumBromide) assay. © 2020 Wiley Periodicals, Inc.