The survival of pathogenic bacteria in the oral cavity depends on their successful adhesion to dental surfaces and their ability to develop into biofilms, known as dental plaque. Bacteria from the dental plaque are responsible for the development of dental caries, gingivitis, periodontitis, stomatitis and peri-implantitis. Certain metal nanoparticles have been suggested for infection control and the management of the oral biofilm. Here, it is shown that application of a silver nano-coating directly on dentine can successfully prevent the biofilm formation on dentine surfaces as well as inhibit bacterial growth in the surrounding media. This silver nano-coating was found to be stable (>98.8%) and to maintain its integrity in biological fluids. Its antibacterial activity was compared to silver nitrate and the widely used clinical antiseptic, chlorhexidine. The bacterial growth and cell viability were quantitatively assessed by measuring the turbidity, proportion of live and dead cells and lactate production. All three bioassays showed that silver nanoparticles and silver nitrate dentine coatings were equally highly bactericidal (>99.5%), while inhibiting bacterial adhesion. However, the latter caused significant dentine discolouration (ΔE* = 50.3). The chlorhexidine coating showed no antibacterial effect. Thus, silver nanoparticles may be a viable alternative to both chlorhexidine and silver nitrate, protecting from dental plaque and secondary caries when applied as a dentine coating, while they may provide the platform for creating anti-biofilm surfaces in medical devices and other biomedical applications.