Hite cell lineage analyzed [14,39]. However, a further study reported differences in the genotoxic D-Fructose-6-phosphate (disodium) salt site effects observed in human lung cancer and macrophage cells just after acute exposure to differently functionalized PSNPs particles [40]. Interestingly, these authors discovered no important genotoxic effects of exposure to pristine, non-functionalized PSNPs. These variations highlight the significance of particle structure in the observed effects, each at acute and long-term endpoints. As for the levels of ROS present in D-Vitamin E acetate Autophagy exposed cells, lots of earlier studies have documented increases in oxidative stress as a result of nanoplastics exposure [21,24,28,40]. Having said that, we did not observe such an effect in our study. Instead, exposed samples showed comparable levels of ROS to unexposed controls each just after the brief and long-term exposure, indicating that exposure to pristine PSNPs did not trigger a rise inside the oxidative tension levels of exposed cells. Overall, the only indication of important stress-related pathways observed was the overexpression of HO1 and SOD2 genes in Caco-2 cells exposed to PSNPs for eight weeks. Nevertheless, we did not identified other evidence of ROS production, DNA harm, or oxidative damage throughout all measured endpoints, suggesting that long-term exposure to pristine PSNPs alone does not induce any of these effects in Caco-2 cells. Even so, environmental weathering of nanoplastic particles may well bring about adjustments to their properties that could alter their toxicity towards exposed organisms, for instance size and surface charge [1,31]. Also, the structural change could alter their adsorption of environmental contaminants, which is already well documented and may possibly induce indirect damaging effects on exposed organisms, specially if accumulated and amplified throughout the meals web [12,41,42]. Therefore, it is actually important to acquire experimental data that improved reflects the realistic conditions of exposure to nanoplastics, as toxicities observed in controlled laboratory experiments, for instance these focusing on acute exposures, might vastly differ from the actual hazard posed by the chronic exposure to weathered and altered nanoplastics. Our study, which proposes a long-term exposure at lower concentrations, aims to greater reflect environmental conditions, and hence give information and facts which is extra alike to the real-case scenario.Author Contributions: C.C., A.H. and R.M. planned the experiments. J.D., M.d.B., A.V., S.P. and C.C. carried out the experimental element. M.d.B., C.C. and J.D. analyzed the data, carried out the statistical evaluation, and prepared the tables/figures. C.C., J.D., A.H. and R.M. wrote the final manuscript. All authors have read and agreed towards the published version in the manuscript. Funding: This project received funding in the European Union’s Horizon 2020 research and innovation program below grant agreement No. 965196. Institutional Assessment Board Statement: Not applicable. Informed Consent Statement: Not applicable. Information Availability Statement: Not applicable. Acknowledgments: J.D. was supported by a Predoctoral Fellowship (PIF) from the Universitat Aut oma de Barcelona. We thank Victor Puntes’ group (Inorganic Nanoparticles Group, ICN2) for supplying the protocol to characterize PS nanoparticles. Conflicts of Interest: The authors declare that they’ve no recognized competing monetary interests or personal relationships that could have appeared to influence the function reported within this paper.
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