AbstractZinc nanoparticles (ZnNPs) are showing promising medical applications. However, their cytotoxicity is relatively high. This study aims to use a green synthesis technique based on the natural propolis (honeybees glue) and produce three counterparts of Zinc nanoparticles (ZnO-NPs, AuNPs@ZnO and AgNPs@ZnO). Then, the three would be analyzed for their potential antibacterial activity and cytotoxicity. The study compares the antibacterial activity and cytotoxicity of ZnO-NPs alone to its combination with gold and silver (AuNPs@ZnO and AgNPs@ZnO). The results showed that AgNPs@ZnO had contributed significantly to antibacterial activity against Gram-positive and Gram-negative bacteria. Moreover, AuNPs@ZnO and AgNPs@ZnO showed similar cytotoxicity to ZnO-NPs with higher safety. Furthermore, the Scanning and Transmission Electron Microscopes’ micrographs (SEM and TEM) displayed the biosynthesized ZnNPs that have a spherical shape with sizes ranging from 17 to 70 nm, from 45 to 75 nm, and from 22 to 73 nm for ZnONPs, ZnO-AgNPs, free AgNPs, and ZnO-AuNPs, respectively. In addition, zeta potential values for ZnONPs were 11.4 mV, while ZnO-AgNPs and ZnO-AuNPs have a higher overall charge of 13.6 mV and 23.9 mV, respectively. Furthermore, ZnNPs exhibited antibacterial activity, yet AgNPs@ZnO demonstrated the highest antibacterial action against Gram-negative and Gram-positive bacteria. In addition, cytotoxicity analyses of biosynthesized ZnNPs on three cell lines; breast cancer cell (MCF-7), liver cancer cell (HepG-2) and Human skin fibroblast (HSF) revealed that Zn-NPs in a combination could function as an anticancer agent. Therefore, there are promising medical applications for the biosynthesized ZnNPs in combinations with other metals, such as ZnO-AgNPs, as safe antibacterial and anticancer medicines. Graphical Abstract