Responses of freshwater biofilm formation processes (from colonization to maturity) to anatase and rutile TiO nanoparticles: Effects of nanoparticles aging and transformation.

Most of the current studies on the toxicology of pristine nanoparticles (NPs) are environmentally irrelevant, because their ''aging'' process accompanied by the physicochemical transformation is inevitable in the environment. Considering aging phenomenon will gain a better understanding of the toxicity and fate of NPs in the environment. Here, we focused on the physicochemical transformation of anatase-NPs (TiO-A) and rutile-NPs (TiO-R) after 90 days of aging and investigated the responses of freshwater biofilm formation to the stress changes of naturally aged TiO-NPs (aTiO-NPs). We found that after aging, the TiO-NPs underwent sophisticated physicochemical transformations in the original morphology and microstructure owing to organic and crystal salts inclusions, such as energy band changes and the formation of Ti on the NPs surfaces. These comprehensive transformations increased the stability of NPs in the exposed suspension. However, the physicochemical transformations were crystal-forms-dependent, and aging did not change the crystal structure and crystallinity. Interestingly, compared to pristine NPs, aTiO-NPs showed much lower cytotoxicity and had the weaker ability to promote or inhibit the biofilm formation (p < 0.05) owing to the passivation of photoactivity caused by the comprehensive effect of the inclusions, especially for aTiO-A. Regardless of aging or not of crystal forms, responses of biofilm formation were exposure-concentration-dependent, namely low concentration promotion (0.1 mg/L) and high concentration inhibition (10 mg/L), e.g., role transition of the pioneers (algae or bacteria) in initial colonization, extracellular polymeric substances (EPS) secretion and compositions of development stages with polysaccharide (PS)-rich and protein (PRO)-rich stages, and biomass and cell activity at different depths of mature biofilms. The reactive oxygen species (ROS) induced by TiO-NPs showed typical hormesis. The changing trends of the autoinducers (c-di-GMP and quorum sensing signals including AHL and AI-2) were highly consistent with the growth stages of biofilms and were stimulated or suppressed by TiO-NPs. The NPs crystal-dependently changed the microorganism community structures, while the UPGMA clustering of bacteria was based on the growth stages of the biofilms. The toxic mechanisms revealed that photoactivity and nanoscale retention of particles are the main reasons for the differences in the ecological stress capacity of four kinds of TiO-NPs. Aging reduced characteristic differences of two pristine NPs and even reversed their relative stresses levels (p > 0.05). However, the toxicity of high-concentration aTiO-NPs (10 mg/L) remained serious in a water environment. This study provides a better understanding for the water environmental risks evaluation and policy control of nanoparticles, that is, the effect of time aging has to be considered.