The increase in the prevalence of bacterial resistance to antibiotics has become one of the main health problems worldwide, thus threatening the era of antibiotics most frequently used in the treatment of infections. The need to develop new therapeutic strategies against multidrug resistant microorganisms, such as the combination of selected antimicrobials, can be considered as a suitable alternative. The in vitro activities of two groups of conventional antimicrobial agents alone and in combination with silver nanoparticles (AgNPs) were investigated against a set of ten multidrug resistant clinical isolate and two references strains by MIC assays and checkerboard testing, as well as their cytotoxicity, which was evaluated on human fibroblasts by MTT assay at the same concentration of the antimicrobial agents alone and in combination. Interesting results were achieved when the AgNPs and their combinations were characterized by Dynamic Light Scattering (DLS), Zeta Potential, Transmission Electron Microscopy (TEM), UV–visible spectroscopy and Fourier Transforms Infrared (FTIR) spectroscopy. The in vitro activities of ampicillin, in combination with AgNPs, against the 12 microorganisms showed one Synergy, seven Partial Synergy and four Additive effects, while the results with amikacin and AgNPs showed three Synergy, eight Partial Synergy and one Additive effects. The cytotoxic effect at these concentrations presented a statistically significant decrease of their cytotoxicity (p < 0.05). These results indicate that infections caused by multidrug resistant microorganisms could be treated using a synergistic combination of antimicrobial drugs and AgNPs. Further studies are necessary to evaluate the specific mechanisms of action, which could help predict undesirable off-target interactions, suggest ways of regulating a drug’s activity, and identify novel therapeutic agents in this health problem.