The present study evaluated the effect of glass application with and without silver-doped soda-lime glass on roughness, biofilm formation, cell viability and flexural strength of a zirconia. Samples of 3-YTZP (3 mol% yttria stabilized tetragonal zirconia polycrystals) were divided into: polished (P); glaze (G); glass infiltration (INF); 4% silver-doped soda-lime glass (Ag4); glass infiltration + 4% silver-doped soda-lime glass (INF-Ag4); 5% silver-doped soda-lime glass (Ag5); glass infiltration + 5% silver-doped soda-lime glass (INF-Ag5). Samples were submitted to the following analyses: roughness (Ra); free surface energy (FSE); colony-forming units count (log CFU/mL); scanning electron microscopy (SEM); cytotoxicity (MTT assay) and flexural strength. Ag5 had greater roughness and FSE, but less biofilm adherence. In the CFU, silver-doped soda-lime glass groups inhibited the growth of Candida albicans, while the Ag5 inhibited Streptococcus mutans and none of the groups was effective against Streptococcus sanguinis. In the qualitative evaluation, lower number of colonies in the Ag5 grew up, compared to the control groups (P; G and INF) for both C. albicans and S. mutans. Regarding the MTT assay, the Ag4, INF-Ag4 and INF-Ag5 obtained percentage of cell viability greater than 50%. Ag5 showed lower flexural strength when compared to the control groups, while the application of glass infiltration increased the flexural strength by formation of a graded region between zirconia-glass. In conclusion, Ag5 had the greatest antimicrobial effect, Ag4 and INF-Ag4 were the less cytotoxic and the INF was the most resistant to fracture. Therefore, INF-Ag4 conciliates the best performance in terms of antimicrobial and mechanical properties for a 3-YTZP.