title: Toxicity evaluation of manufactured CeO2 nanoparticles before and after alteration: combined physicochemical and whole-genome expression analysis in Caco-2 cells.
abstract: BACKGROUND
  Engineered nanomaterials may release nanosized residues, by degradation, throughout their life cycle. These residues may be a threat for living organisms. They may be ingested by humans through food and water. Although the toxicity of pristine CeO2 nanoparticles (NPs) has been documented, there is a lack of studies on manufactured nanoparticles, which are often surface modified. Here, we investigated the potential adverse effects of CeO2 Nanobyk 3810™ NPs, used in wood care, and their residues, altered by light or acid.
  
  
  RESULTS
  Human intestinal Caco-2 cells were exposed to residues degraded by daylight or in a medium simulating gastric acidity. Size and zeta potential were determined by dynamic light scattering. The surface structure and redox state of cerium were analyzed by transmission electronic microscopy (TEM) and X-ray absorption spectroscopy, respectively. Viability tests were performed in Caco-2 cells exposed to NPs. Cell morphology was imaged with scanning electronic microscopy. Gene expression profiles obtained from cells exposed to NPs before and after their alteration were compared, to highlight differences in cellular functions.No change in the cerium redox state was observed for altered NPs. All CeO2 NPs suspended in the culture medium became microsized. Cytotoxicity tests showed no toxicity after Caco-2 cell exposure to these various NPs up to 170 μg/mL (24 h and 72 h). Nevertheless, a more-sensitive whole-gene-expression study, based on a pathway-driven analysis, highlighted a modification of metabolic activity, especially mitochondrial function, by altered Nanobyk 3810™. The down-regulation of key genes of this pathway was validated by qRT-PCR. Conversely, Nanobyk 3810™ coated with ammonium citrate did not display any adverse effect at the same concentration.
  
  
  CONCLUSIONS
  The degraded nanoparticles were more toxic than their coated counterparts. Desorption of the outside layer was the most likely cause of this discrepancy in toxicity. It can be assumed that the safe design of engineered nanoparticles could include robust protective layers conferring on them greater resistance to alteration during their life cycle.
authors.forename: Matthieu
authors.initials: M
authors.lastname: Fisichella
authors.forename: Frederic
authors.initials: F
authors.lastname: Berenguer
authors.forename: Gerard
authors.initials: G
authors.lastname: Steinmetz
authors.forename: Melanie
authors.initials: M
authors.lastname: Auffan
authors.forename: Jerome
authors.initials: J
authors.lastname: Rose
authors.affiliation: CEA, IBEB, SBTN, Laboratoire d'Etude des Protéines Cibles, F-30207 Bagnols-sur-Cèze, France. odette.prat@cea.fr.
authors.forename: Odette
authors.initials: O
authors.lastname: Prat
mesh_terms: D018938:Caco-2 Cells; D048430:Cell Shape / Q000187:drug effects; D002563:Cerium / Q000737:chemistry / Q000633:toxicity*; D045222:Electron Transport Chain Complex Proteins / Q000235:genetics / Q000378:metabolism; D015894:Genome, Human; D006801:Humans; D053758:Nanoparticles / Q000737:chemistry / Q000633:toxicity* / Q000648:ultrastructure; D020411:Oligonucleotide Array Sequence Analysis; D010316:Particle Size; D059467:Transcriptome / Q000187:drug effects*
chemical_list: D045222:Electron Transport Chain Complex Proteins; D002563:Cerium; C030583:ceric oxide