The aim of this study was to verify the possible physical and chemical changes that may occur on the surface of Ag–TiCN coatings after exposure to the culture media used in microbiological and cytotoxic assays, respectively tryptic soy broth (TSB) and Dulbecco's modified eagle's medium (DMEM). After sample immersion for 24 h in the media, analyses were performed by glow discharge optical emission spectroscopy discharge radiation (GDOES), Rutherford backscattering spectroscopy (RBS) and x-ray photoelectron spectroscopy (XPS). The results of GDOES profile, RBS and XPS spectra, of samples immersed in TSB, demonstrated the formation of a thin layer of carbon, oxygen and nitrogen that could be due to the presence of proteins in TSB. After 24 h of immersion in DMEM, the results showed the formation of a thin layer of calcium phosphates on the surface, since the coatings displayed a highly oxidized surface in which calcium and phosphorus were detected. All these results suggested that the formation of a layer on the coating surface prevented the release of silver ions in concentrations that allow antibacterial activity.IC acknowledges the financial support of FCT-Fundacao para a Ciencia e a Tecnologia through the grant SFRH/BD/67022/2009. REG acknowledges support from Ramon y Cajal programme (RyC2007-0026). This research is sponsored by FEDER funds through the program COMPETE-Programa Operacional Factores de Competitividade and by national funds through FCT-Fundacao para a Ciencia e a Tecnologia in the framework of the Strategic Projects PESTC/FIS/UI607/2011, PEST-C/EME/UI0285/2011, PTDC/CTM/102853/2008.The authors thank the FCT Strategic Project PEst-OE/EQB/LA0023/2013 and the Project 'BioHealth-Biotechnology and Bioengineering approaches to improve health quality', Ref. NORTE-07-0124-FEDER-000027, co-funded by the Programa Operacional Regional do Norte (ON.2-O Novo Norte), QREN, FEDER. The authors also thank the project 'Consolidating Research Expertise and Resources on Cellular and Molecular Biotechnology at CEB/IBB', Ref. FCOMP-01-0124-FEDER-027462
