The aim of this study was to develop a nanostructured bioactive surface based on zinc oxide, sodium stearate (C18) and usnic acid (UA) exhibiting harmless effects with respect to the human cells, but with a significant antimicrobial effect, limiting the attachment and biofilm formation of food pathogens. ZnO nanoparticles were synthesized by sol-gel method and functionalized with C18 and UA. The coatings were fabricated by matrix assisted pulsed laser evaporation technique (MAPLE) and further characterized by TEM, SEM, SAED, XRD and IRM. The biological characterization of the prepared coatings consisted in cytotoxicity and antimicrobial assays. The cytotoxicity of ZnO@C-18 and ZnO@C-18-UA films was evaluated with respect to the human skin fibroblasts (CCD 1070SK cell line) by phase contrast microscopy, MTT assay and nitric oxide (NO) release. The covered surfaces exhibited a decreased cell attachment, effect which was more pronounced in the presence of UA as shown by purple formazan staining of adhered cells. The unattached fibroblasts remained viable after 24 h in the culture media as it was revealed by their morphology analysis and NO level which were similar to uncovered slides. The quantitative microbiological assays results have demonstrated that the bioactive coatings have significantly inhibited the adherence and biofilm formation of Salmonella enterica. The obtained results recommend these materials as efficient approaches in developing anti-adherent coatings for various industrial, medical and food processing applications. (C) 2015 Elsevier B.V. All rights reserved.