Cell adhesion to biomaterials and cell proliferation are affected, among others, by the chemical composition and surface microtopography of the biomaterial. These are, in turn, influenced by sterilisation processes. The aim of this study was to evaluate the sensitivity of osteoblasts and fibroblasts to TiN surfaces deprived of an oxide film or with an enhanced oxide film due to sterilisation in plasma or in an autoclave, respectively. Type TiN+T2N+alpha Ti(N) nitrided surface layers were produced under glow discharge conditions and sterilized in plasma Sterrad 100 and in a steam autoclave. The results provide evidence that nitrided surface layers improved wear-and corrosion, resistance of the titanium alloy and that these features were not influenced by sterilisation procedures that resulted in various oxide films. In vitro study of human fibroblast and osteoblast-like cells cultured on TiN surfaces modified by sterilisation processes showed that cells respond to the presence of an enhanced oxide film by aggregating, exhibiting lower viability and increased apoptosis, while to a surface deprived of oxides, by regular spreading and high viability. It can be concluded that differences in surface composition and topography of the nitrided surface layers due to various types of sterilisation elicit specific cell responses, and plasma sterilisation seems to be optimal for bone implants with this surface layer.