title: High-throughput, quantitative assessment of the effects of low-dose silica nanoparticles on lung cells: grasping complex toxicity with a great depth of field.
abstract: BACKGROUND
  The toxicity of manufactured fumed silica nanoparticles (NPs) remains poorly investigated compared to that of crystalline silica NPs, which have been associated with lung diseases after inhalation. Amorphous silica NPs are a raw material for manufactured nanocomposites, such as cosmetics, foods, and drugs, raising concerns about their potential toxicity.
  
  
  RESULTS
  The size of the NPs was determined by dynamic light scattering and their shape was visualized by atomic force microscopy (10 ± 4 nm). The pertinent toxicological concentration and dynamic ranges were determined using viability tests and cellular impedance. We combined transcriptomics and proteomics to assess the cellular and molecular effects of fumed silica in A549 human alveolar epithelial cells. The "no observed transcriptomic adverse effect level" (NOTEL) was set to 1.0 μg/cm(2), and the "lowest observed adverse transcriptional effect level" (LOTEL) was set at 1.5 μg/cm(2). We carried out genome-wide expression profiles with microarrays and identified, by shotgun proteomics, the exoproteome changes in lung cells after exposure to NP doses (0.1, 1.0, 1.5, 3.0, and 6.0 μg/cm(2)) at two time points (24 h and 72 h). The data revealed a hierarchical, dose-dependent cellular response to silica NPs. At 1.5 μg/cm(2), the Rho signaling cascade, actin cytoskeleton remodeling, and clathrin-mediated endocytosis were induced. At 3.0 μg/cm(2), many inflammatory mediators were upregulated and the coagulation system pathway was triggered. Lastly, at 6.0 μg/cm(2), oxidative stress was initiated. The proteins identified in the extracellular compartment were consistent with these findings.
  
  
  CONCLUSIONS
  The alliance of two high-throughput technologies allowed the quantitative assessment of the cellular effects and molecular consequences of exposure of lung cells to low doses of NPs. These results were obtained using a pathway-driven analysis instead of isolated genes. As in photography, toxicogenomics allows, at the same time, the visualization of a wide spectrum of biological responses and a "zoom in" to the details with a great depth of field. This study illustrates how such an approach based on human cell culture models is a valuable predictive screening tool to evaluate the toxicity of many potentially harmful emerging substances, alone or in mixtures, in the framework of future regulatory reinforcements.
authors.affiliation: CEA, IBEB, SBTN, F-30207, Bagnols-sur-Cèze, France. cedric.pisani@cea.fr.
authors.forename: Cédric
authors.initials: C
authors.lastname: Pisani
authors.affiliation: CEA, IBEB, SBTN, F-30207, Bagnols-sur-Cèze, France. jean-charles.gaillard@cea.fr.
authors.forename: Jean-Charles
authors.initials: JC
authors.lastname: Gaillard
authors.affiliation: CEA, IBEB, SBTN, F-30207, Bagnols-sur-Cèze, France. virginie.nouvel@cea.fr.
authors.forename: Virginie
authors.initials: V
authors.lastname: Nouvel
authors.affiliation: CEA, IBEB, SBTN, F-30207, Bagnols-sur-Cèze, France. michael.odorico@cea.fr.
authors.forename: Michaël
authors.initials: M
authors.lastname: Odorico
authors.affiliation: CEA, IBEB, SBTN, F-30207, Bagnols-sur-Cèze, France. jean.armengaud@cea.fr.
authors.forename: Jean
authors.initials: J
authors.lastname: Armengaud
authors.affiliation: CEA, IBEB, SBTN, F-30207, Bagnols-sur-Cèze, France. odette.prat@cea.fr.|International Consortium for the Environmental Implications of Nanotechnology (iCEINT), Aix-en-Provence, France. odette.prat@cea.fr.
authors.forename: Odette
authors.initials: O
authors.lastname: Prat
mesh_terms: D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism; D008168:Lung / Q000166:cytology; D053758:Nanoparticles / Q000633:toxicity*; D018384:Oxidative Stress / Q000187:drug effects; D010316:Particle Size; D020543:Proteome / Q000032:analysis / Q000378:metabolism; D012822:Silicon Dioxide / Q000737:chemistry*; D053719:Tandem Mass Spectrometry; D059467:Transcriptome / Q000187:drug effects*; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D018836:Inflammation Mediators; D020543:Proteome; D012822:Silicon Dioxide