In this study, we aimed to evaluate the antibacterial and antifungal properties, cytotoxicity, and elution behavior of copper oxide (CuO) thin films with varying concentrations and roughness values. CuO films greater than 0.2 mol % showed 99.9 % antimicrobial activity against Escherichia coli, Staphylococcus aureus, Campylobacter jejuni, and Penicillium funiculosum. Cu ions were found to be noncytotoxic in New Zealand white rabbits. The concentration of Cu ions from CuO thin films eluted in drinking water in 24 h at 100 °C was 0.014 μg L(-1), which was below the standard acceptable level of 0.02 μg L(-1). The transmittance of CuO thin film-coated glass was similar to that of parent glass. The antimicrobial activity, cytotoxicity, elution behavior, and transmittance of CuO deposited on glass suggest that these films could be useful in household devices and display devices.