Time-resolved toxicity study reveals the dynamic interactions between uncoated silver nanoparticles and bacteria

It is still unclear whether the toxicity of silver nanoparticles (AgNPs) can be attributed solely to the release of Ag⁺ or whether dissolved and nanoparticulate Ag act in parallel; this is due to the difficulty in distinguishing Ag⁺- from AgNP-effects. Also, AgNPs undergo changes during toxicity tests. This is the first study to investigate the influence of AgNP dissolution over time on viable counts at high time resolution and low cell density, avoiding the apparently reduced toxicity at higher cell densities identified in our study. Uncapped AgNPs were synthesized to avoid any interference from surface coatings. The transformations of AgNPs during storage were reduced. Lowering the concentration of AgNPs reduced their aggregation in Davis minimal medium (DMM). Also, AgNPs dissolved more slowly in DMM than in water. The minimum inhibitory concentrations (MICs) of Ag⁺ and AgNPs increased with cell density according to a power law, suggesting that binding to cells decreased effective concentrations. However, AgNPs acted as a reservoir of Ag, releasing new Ag⁺ to maintain the Ag stress. The toxicity of AgNPs was dominated by dissolved Ag. Combining controlled conditions, high time-resolution and low cell density, we could demonstrate different roles of ionic and nano Ag in bacterial death caused by AgNPs.