Plastic nanoparticles interfere with extracellular vesicle pathway in primary astrocytes.

The extensive production and use of plastics in recent decades has led to environmental pollution. It has been discovered that plastic microparticles (MPs) and nanoparticles (NPs), formed under the influence of physical forces, can pose a significant health risk. Increasing evidence indicates that NPs can have various toxic effects, including oxidative stress and cell death. However, the mechanisms underlying their toxicity are still under investigation. In this study, we examined whether polystyrene nanoparticles (PS-NPs) are internalized in primary astrocytes. We tracked their intracellular fate and search for potential interference with the intercellular communication pathway mediated by extracellular vesicles (EVs). Primary astrocyte cultures were exposed to fluorescent PS-NPs at concentrations of 0.5, 1, 25 and 50 µg/mL for 24, 48 and 72 hours. Based on electron microscopic analysis and confocal imaging, we determined that PS-NPs are internalized in astrocytes and accumulate in the cytoplasm in a concentration-dependent manner, localizing to endosomal-lysosomal system. Astrocytes exposed to PS-NPs form EVs containing encapsulated PS-NPs, which are released into the culture medium after 72 h of exposure and can be transferred via this route to other cells. As shown by proteomic analysis, PS-NPs affects the composition of the protein cargo of released EVs by decreasing the representation of proteins such as CD47, CSTB and CNDP2. Intercellular transport of PS-NPs in primary astrocytes is mediated by EVs system. EV-mediated release of PS-NPs may alleviate their toxicity in a single astrocyte but may also contribute to the spread of their toxic effect to neighbouring astrocytes. Exposure to PS-NPs interferes with the mechanism of protein sorting, thereby potentially influencing the EV-mediated cell-cell communication pathway.