title: Specific uptake and genotoxicity induced by polystyrene nanobeads with distinct surface chemistry on human lung epithelial cells and macrophages.
abstract: Nanoparticle surface chemistry is known to play a crucial role in interactions with cells and their related cytotoxic effects. As inhalation is a major route of exposure to nanoparticles, we studied specific uptake and damages of well-characterized fluorescent 50 nm polystyrene (PS) nanobeads harboring different functionalized surfaces (non-functionalized, carboxylated and aminated) on pulmonary epithelial cells and macrophages (Calu-3 and THP-1 cell lines respectively). Cytotoxicity of in mass dye-labeled functionalized PS nanobeads was assessed by xCELLigence system and alamarBlue viability assay. Nanobeads-cells interactions were studied by video-microscopy, flow cytometry and also confocal microscopy. Finally ROS generation was assessed by glutathione depletion dosages and genotoxicity was assessed by γ-H2Ax foci detection, which is considered as the most sensitive technique for studying DNA double strand breaks. The uptake kinetic was different for each cell line. All nanobeads were partly adsorbed and internalized, then released by Calu-3 cells, while THP-1 macrophages quickly incorporated all nanobeads which were located in the cytoplasm rather than in the nuclei. In parallel, the genotoxicity study reported that only aminated nanobeads significantly increased DNA damages in association with a strong depletion of reduced glutathione in both cell lines. We showed that for similar nanoparticle concentrations and sizes, aminated polystyrene nanobeads were more cytotoxic and genotoxic than unmodified and carboxylated ones on both cell lines. Interestingly, aminated polystyrene nanobeads induced similar cytotoxic and genotoxic effects on Calu-3 epithelial cells and THP-1 macrophages, for all levels of intracellular nanoparticles tested. Our results strongly support the primordial role of nanoparticles surface chemistry on cellular uptake and related biological effects. Moreover our data clearly show that nanoparticle internalization and observed adverse effects are not necessarily associated.
authors: Institut National de l'Environnement Industriel et des Risques (INERIS), Unité de Toxicologie Expérimentale, Parc Technologique ALATA, BP2, Verneuil-en-Halatte, France.
authors: Vincent
authors: V
authors: Paget
authors: Institut National de l'Environnement Industriel et des Risques (INERIS), Unité de Toxicologie Expérimentale, Parc Technologique ALATA, BP2, Verneuil-en-Halatte, France; Laboratoire de chimie et toxicologie analytique et cellulaire (C-TAC) / UMR CNRS 8638, Faculté de Pharmacie de Paris, Université Paris Descartes (PRES Sorbonne Paris Cité), Paris, France.
authors: Samir
authors: S
authors: Dekali
authors: CEA, DSV, iRCM, Plateforme imagerie photonique, Fontenay-aux-Roses, France.
authors: Thierry
authors: T
authors: Kortulewski
authors: CEA, DSV, iRCM, Laboratoire Cancérologie Expérimentale (LCE), Fontenay-aux-Roses, France.
authors: Romain
authors: R
authors: Grall
authors: Institut National de l'Environnement Industriel et des Risques (INERIS), Unité de Toxicologie Expérimentale, Parc Technologique ALATA, BP2, Verneuil-en-Halatte, France.
authors: Christelle
authors: C
authors: Gamez
authors: Institut National de l'Environnement Industriel et des Risques (INERIS), Unité de Toxicologie Expérimentale, Parc Technologique ALATA, BP2, Verneuil-en-Halatte, France.
authors: Kelly
authors: K
authors: Blazy
authors: Institut National de l'Environnement Industriel et des Risques (INERIS), Unité de Toxicologie Expérimentale, Parc Technologique ALATA, BP2, Verneuil-en-Halatte, France.
authors: Olivier
authors: O
authors: Aguerre-Chariol
authors: CEA, DSV, iRCM, Laboratoire Cancérologie Expérimentale (LCE), Fontenay-aux-Roses, France.
authors: Sylvie
authors: S
authors: Chevillard
authors: Institut National de l'Environnement Industriel et des Risques (INERIS), Unité de Toxicologie Expérimentale, Parc Technologique ALATA, BP2, Verneuil-en-Halatte, France.
authors: Anne
authors: A
authors: Braun
authors: Laboratoire de chimie et toxicologie analytique et cellulaire (C-TAC) / UMR CNRS 8638, Faculté de Pharmacie de Paris, Université Paris Descartes (PRES Sorbonne Paris Cité), Paris, France.
authors: Patrice
authors: P
authors: Rat
authors: Institut National de l'Environnement Industriel et des Risques (INERIS), Unité de Toxicologie Expérimentale, Parc Technologique ALATA, BP2, Verneuil-en-Halatte, France.
authors: Ghislaine
authors: G
authors: Lacroix
mesh_terms: D000586:Amination; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D053903:DNA Breaks, Double-Stranded / Q000187:drug effects*; D004249:DNA Damage; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D006657:Histones / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008264:Macrophages / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D053758:Nanoparticles / Q000737:chemistry / Q000633:toxicity* / Q000648:ultrastructure; D010078:Oxazines; D010316:Particle Size; D011137:Polystyrenes / Q000737:chemistry / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000819:agonists / Q000378:metabolism; D013499:Surface Properties; D014966:Xanthenes
chemical_list: C468783:H2AX protein, human; D006657:Histones; D010078:Oxazines; D011137:Polystyrenes; D017382:Reactive Oxygen Species; D014966:Xanthenes; C005843:resazurin; D005978:Glutathione