Sonicated polyethylene terephthalate nano- and micro-plastic-induced inflammation, oxidative stress, and autophagy in vitro.

The environmental presence of nano- and micro-plastic particles (NMPs) is suspected to have a negative impact on human health. Environmental NMPs are difficult to sample and use in life science research, while commercially available plastic particles are too morphologically uniform. Additionally, this NMPs exposure exhibited biological effects, including cell internalization, oxidative stress, inflammation, cellular adaptation, and genotoxicity. Therefore, developing new methods for producing heterogenous NMPs as observed in the environment is important as reference materials for research. Thus, we aimed to generate and characterize NMPs suspensions using a modified ultrasonic protocol and to investigate their biological effects after exposure to different human cell lines. To this end, we produced polyethylene terephthalate (PET) NMPs suspensions and characterized the particles by dynamic light scattering and scanning electron microscopy. Ultrasound treatment induced polymer degradation into smaller and heterogeneous PET NMPs shape fragments with similar surface chemistry before and after treatment. A polydisperse suspension of PET NMPs with 781 nm in average size and negative surface charge was generated. Then, the PET NMPs were cultured with two human cell lines, A549 (lung) and HaCaT (skin), addressing inhalation and topical exposure routes. Both cell lines interacted with and have taken up PET NMPs as quantified via cellular granularity assay. A549 but not HaCaT cell metabolism, viability, and cell death were affected by PET NMPs. In HaCaT keratinocytes, large PET NMPs provoked genotoxic effects. In both cell lines, PET NMPs exposure affected oxidative stress, cytokine release, and cell morphology, independently of concentration, which we could relate mechanistically to Nrf2 and autophagy activation. Collectively, we present a new PET NMP generation model suitable for studying the environmental and biological consequences of exposure to this polymer.

Lipid Corona Formation on Micro- and Nanoplastic Particles Modulates Uptake and Toxicity in A549 Cells.

Plastic waste is a global issue leaving no continents unaffected. In the environment, ultraviolet radiation and shear forces in water and land contribute to generating micro- and nanoplastic particles (MNPP), which organisms can easily take up. Plastic particles enter the human food chain, and the accumulation of particles within the human body is expected. Crossing epithelial barriers and cellular uptake of MNPP involves the interaction of plastic particles with lipids. To this end, we generated unilamellar vesicles from POPC (1-palmitoyl-2-oleoyl-glycero-3-phosphocholine) and POPS (1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine) and incubated them with pristine, carboxylated, or aminated polystyrene spheres (about 1 µm in diameter) to generate lipid coronas around the particles. Lipid coronas enhanced the average particle sizes and partially changed the MNPP zeta potential and polydispersity. In addition, lipid coronas led to significantly enhanced uptake of MNPP particles but not their cytotoxicity, as determined by flow cytometry. Finally, adding proteins to lipid corona nanoparticles further modified MNPP uptake by reducing the uptake kinetics, especially in pristine and carboxylated plastic samples. In conclusion, our study demonstrates for the first time the impact of different types of lipids on differently charged MNPP particles and the biological consequences of such modifications to better understand the potential hazards of plastic exposure.