The neurodevelopmental toxicity induced by combined exposure of nanoplastics and penicillin in embryonic zebrafish: The role of aging processes

As ubiquitous contaminants, nanoplastics and antibiotics are frequently co-presence and widely detected in the freshwater environment and biota, posing a high co-exposure risk to aquatic organisms and even humans. More importantly, how the aging process of nanoplastics affects the joint toxic potential of nanoplastics and antibiotics has not been explored. Here, we generated two aged polystyrene nanoplastics (PS) by UV radiation (UV-PS) and ozonation (O3-PS). Non-teratogenic concentrations of pristine PS (80 nm) and antibiotics penicillin (PNC) coexposure synergistically suppressed the embryo heart beating and behaviors of spontaneous movement, touch response, and larval swimming behavioral response. Pristine PS and aged UV-PS, but not aged O3-PS, showed similar effects on zebrafish embryo/larval neurodevelopment. However, when co-exposure with PNC, both aged PS, but not pristine PS, showed antagonistic effects. In late-stage juvenile social behavior testing, we found that PS decreased the exploration in light/dark preference assay. The synergistic effect of aged PS with PNC was further explored, including cellular apoptosis, ROS formation, and neurotransmitter metabolite regulation. Mechanistically, aged UV-PS but not O3-PS significantly increased the adsorption rate of PNC compared to pristine PS, which may account for the toxicity difference between the two aged PS. In conclusion, our results confirmed that PS served as a carrier for PNC, and the environmental aging process changed their neurobehavioral toxicity pattern in vivo.