| Summary: | The antibacterial performance and cytotoxic examination of in situ prepared silver nanoparticles (Ag NPs), on inorganic-organic hybrid nanopowder consisting of zirconium dioxide nanoparticles (ZrO<sub>2</sub> NPs) and dihydroquercetin (DHQ), was performed against Gram (−) bacteria <i>Escherichia coli</i> and Gram (+) bacteria <i>Staphylococcus aureus</i>, as well as against human cervical cancer cells HeLa and healthy MRC-5 human cells. The surface modification of ZrO<sub>2</sub> NPs, synthesized by the sol-gel method, with DHQ leads to the interfacial charge transfer (ICT) complex formation indicated by the appearance of absorption in the visible spectral range. The prepared samples were thoroughly characterized (TEM, XRD, reflection spectroscopy), and, in addition, the spectroscopic observations are supported by the density functional theory (DFT) calculations using a cluster model. The concentration- and time-dependent antibacterial tests indicated a complete reduction of bacterial species, <i>E. coli</i> and <i>S. aureus</i>, for all investigated concentrations of silver (0.10, 0.25, and 0.50 mg/mL) after 24 h of contact. On the other side, the functionalized ZrO<sub>2</sub> NPs with DHQ, before and after deposition of Ag NPs, do not display a significant decrease in the viability of HeLa MRC-5 cells in any of the used concentrations compared to the control.
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