Staphylococci coagulase-negative and Staphylococcus aureus are the most common cause of nosocomial infections and can induce a wide spectrum of diseases associated with remarkable morbidity and mortality. Biofilms of staphylococci are intrinsically resistant to conventional antibiotics and, being commonly associated with implanted medical device, such as catheters and vascular grafts, have the potential to cause blood-stream infections. In recent years increasing attention has been addressed to Solid Lipid Nanoparticles (SLNs), thanks to their biodegradability and ability to entrap a variety of biologically active compounds, in the area of modified drug delivery technology in the attempt to overcome drawbacks due to use of the conventional dosage forms. The paper describes the preparation and characterization of vancomycin-loaded SLNs and in vitro studies of the inhibition of Staphylococcus aureus biofilms in the presence of this nanoparticle system. The vancomycin-loaded SLNs were prepared by microemulsion method. Physico-chemical and technological characterization of SLNs was performed concerning drug loading amount, particle size and zeta potential measurements. In vitro microbiological studies, carried out by MTT method. showed that the vancomycin-loaded SLNs have a better activity in the inhibition of Staphylococcus aureus biofilms respect to free vancomycin.