Zinc-substituted cobalt ferrite nanopowders were prepared via a sol-gel route using citric acid as a chelating agent. The influence of zinc concentration on the microstructure, crystal structure, surface wettability, surface roughness, and antibacterial property of zinc-substituted cobalt ferrite nanopowders was investigated systematically. The substitution of zinc influences slightly the microstructure, surface wettability, surface roughness, and crystal structure but strongly affects the antibacterial property of the cobalt ferrite nanopowders.