Nanoplastics (NPs) continue to accumulate in global aquatic and terrestrial systems, posing a potential threat to human health through the food chain and/or other pathways. Both in vivo and in vitro studies have confirmed that the liver is one of the main organs targeted for the accumulation of NPs in living organisms. However, whether exposure to NPs induces size-dependent disorders of liver lipid metabolism remains controversial, and the reversibility of NPs-induced hepatotoxicity is largely unknown. In this study, the effects of long-term exposure to environmentally relevant doses of polystyrene nanoplastics (PS-NPs) on lipid accumulation were investigated in terms of autophagy and lysosomal mechanisms. The findings indicated that hepatic lipid accumulation was more pronounced in mice exposed to 100 nm PS-NPs compared to 500 nm PS-NPs. This effect was effectively alleviated after 50 days of self-recovery for 100 nm and 500 nm PS-NPs exposure. Mechanistically, although PS-NPs exposure activated autophagosome formation through ERK (mitogen-activated protein kinase 1)/mTOR (mechanistic target of rapamycin kinase) signaling pathway, the inhibition of Rab7 (RAB7, member RAS oncogene family), CTSB (cathepsin B), and CTSD (cathepsin D) expression impaired lysosomal function, thereby blocking autophagic flux and contributing to hepatic lipid accumulation. After termination of PS-NPs exposure, lysosomal exocytosis was responsible for the clearance of PS-NPs accumulated in lysosomes. Furthermore, impaired lysosomal function and autophagic flux inhibition were effectively alleviated. This might be the main reason for the alleviation of PS-NPs-induced lipid accumulation after recovery. Collectively, we demonstrate for the first time that lysosomes play a dual role in the persistence and reversibility of hepatotoxicity induced by environmental relevant doses of NPs, which provide novel evidence for the prevention and intervention of liver injury associated with nanoplastics exposure.
Characterization of the cross-sectional morphologies of polymeric membranes are critical in understanding the relationship of structure and membrane separation performances. However, preparation of cross-sectional samples with flat surfaces for scanning electron microscopy (SEM) characterizations is challenging due to the toughness of the non-woven fabric support. In this work, a new frozen section technique was developed to prepare the cross-sectional membrane samples. A special mold was self-designed to embed membranes orientationally. The frozen section parameters, including the embedding medium, cryostat working temperature, and sectioning thickness were optimized. The SEM characterizations demonstrated that the frozen section technique, using ultrapure water as the embedding medium at a working temperature of −30 °C and a sectioning thickness of 0.5 µm, was efficient for the preparation of the membrane samples. Three methods of preparation for the cross-sectional polymeric membranes, including the conventional liquid nitrogen cryogenic fracture, the broad ion beam (BIB) polishing, and the frozen section technique were compared, which showed that the modified frozen section method was efficient and low cost. This developed method could not only accelerate the development of membrane technology but also has great potential for applications in preparation of other solid samples.