Silver nanoparticles exert concentration-dependent influences on biofilm development and architecture

Objectives To investigate the impact of silver nanoparticles (AgNPs) on the biofilm growth and architecture. Materials and methods Silver nitrate was reduced by d-maltose to prepare AgNPs in the presence of ammonia and sodium hydroxide. The physicochemical properties of AgNPs were characterized by transmission electron microscopy, ultraviolet-visible spectroscopy and inductively coupled plasma mass spectrometry. The development of biofilm with and without AgNPs was explored by crystal violet stain. The structures of mature biofilm were visually studied by confocal laser scanning microscopy and scanning electron microscopy. Bacterial cell, polysaccharide and protein within biofilm were assessed quantitatively by colony-counting method, phenol-sulphuric acid method and Bradford assay, respectively. Results The spherical AgNPs (about 30 nm) were successfully synthesized. The effect of AgNPs on Pseudomonas aeruginosa biofilm development was concentration-dependent. Biofilm was more resistant to AgNPs than planktonic cells. Low doses of AgNPs exposure remarkably delayed the growth cycle of biofilm, whereas high concentration (18 mu g/mL) of AgNPs fully prevented biofilm development. The analysis of biofilm architecture at the mature stage demonstrated that AgNPs exposure at all concentration led to significant decrease of cell viability within treated biofilms. However, sublethal doses of AgNPs increased the production of both polysaccharide and protein compared to control, which significantly changed the biofilm structure. Conclusions AgNPs exert concentration-dependent influences on biofilm development and structure, which provides new insight into the role of concentration played in the interaction between antibacterial nanoparticles and biofilm, especially, an ignored sublethal concentration associated with potential unintended consequences.