title: The modality of cell-particle interactions drives the toxicity of nanosized CuO and TiO₂ in human alveolar epithelial cells.
abstract: Metal oxide NPs are abundantly produced in nanotech industries and are emitted in several combustion processes, suggesting the need to characterize their toxic impact on the human respiratory system. The acute toxicity and the morphological changes induced by copper oxide and titanium dioxide NPs (nCuO and nTiO₂) on the human alveolar cell line A549 are here investigated. Cell viability and oxidative stress have been studied in parallel with NP internalization and cell ultrastructural modifications. TiO₂ NPs were abundantly internalized by cells through the endocytic pathway, even they did not induce cell death and ultrastructural lesions. Only after 24h cells were affected by an abundant NP internalization presenting a consequent altered morphology. High cytotoxicity, oxidative stress and severe ultrastructural damages were produced by nCuO, since cell membrane and mitochondria resulted to be heavily affected, even at early exposure time. nCuO-induced toxicity has been interpreted as a consequence of both NPs reactivity and copper ions dissolution in lysosomal compartments, even the free NPs, scattered throughout all the cell compartments, might contribute to the toxicity. The antioxidant N-acetylcysteine was effective in recovering nCuO exposed cells viability and Bafilomycin A1 inhibited copper ions release in phagolysosomes and significantly rescued cells, suggesting a relevant cytotoxic mechanism relative to oxidative damages and authophagic cell death, together with NP internalization and dissolution. Our results support the previous data reporting CuO NPs are highly cytotoxic and genotoxic, and associate their toxic effects with their cell penetration and interaction with various compartments. In conclusion, the so-called "Trojan horse" mechanism and autophagy, are involved in nCuO-induced cell death, even a further research is needed to explain the events occurring at early exposure time.
authors.affiliation: Department of Earth and Environmental Sciences, POLARIS Research Centre, University of Milano Bicocca, 1 piazza della Scienza, 20126 Milan, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Moschini
authors.forename: Maurizio
authors.initials: M
authors.lastname: Gualtieri
authors.forename: Miriam
authors.initials: M
authors.lastname: Colombo
authors.forename: Umberto
authors.initials: U
authors.lastname: Fascio
authors.forename: Marina
authors.initials: M
authors.lastname: Camatini
authors.forename: Paride
authors.initials: P
authors.lastname: Mantecca
mesh_terms: D056809:Alveolar Epithelial Cells / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure; D000975:Antioxidants / Q000494:pharmacology; D001343:Autophagy / Q000187:drug effects*; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000648:ultrastructure; D002470:Cell Survival / Q000187:drug effects; D055598:Chemical Phenomena; D003300:Copper / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D004705:Endocytosis / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism; D008247:Lysosomes / Q000187:drug effects / Q000648:ultrastructure; D053768:Metal Nanoparticles / Q000737:chemistry / Q000633:toxicity*; D008928:Mitochondria / Q000187:drug effects / Q000648:ultrastructure; D016877:Oxidants / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D018384:Oxidative Stress / Q000187:drug effects; D052638:Particulate Matter / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D006180:Proton-Translocating ATPases / Q000494:pharmacology; D014025:Titanium / Q000737:chemistry / Q000633:toxicity*
chemical_list: D000975:Antioxidants; C508606:CXCL8 protein, human; D004791:Enzyme Inhibitors; D016209:Interleukin-8; D016877:Oxidants; D052638:Particulate Matter; C009495:titanium dioxide; D003300:Copper; D014025:Titanium; D006180:Proton-Translocating ATPases; C030973:cupric oxide
keywords: A549; Autophagy; Copper oxide; Microscopy; Nanoparticles; Titanium dioxide