title: MWCNTs of different physicochemical properties cause similar inflammatory responses, but differences in transcriptional and histological markers of fibrosis in mouse lungs.
abstract: Multi-walled carbon nanotubes (MWCNTs) are an inhomogeneous group of nanomaterials that vary in lengths, shapes and types of metal contamination, which makes hazard evaluation difficult. Here we present a toxicogenomic analysis of female C57BL/6 mouse lungs following a single intratracheal instillation of 0, 18, 54 or 162 μg/mouse of a small, curled (CNT(Small), 0.8 ± 0.1 μm in length) or large, thick MWCNT (CNT(Large), 4 ± 0.4 μm in length). The two MWCNTs were extensively characterized by SEM and TEM imaging, thermogravimetric analysis, and Brunauer-Emmett-Teller surface area analysis. Lung tissues were harvested 24h, 3 days and 28 days post-exposure. DNA microarrays were used to analyze gene expression, in parallel with analysis of bronchoalveolar lavage fluid, lung histology, DNA damage (comet assay) and the presence of reactive oxygen species (dichlorodihydrofluorescein assay), to profile and characterize related pulmonary endpoints. Overall changes in global transcription following exposure to CNT(Small) or CNT(Large) were similar. Both MWCNTs elicited strong acute phase and inflammatory responses that peaked at day 3, persisted up to 28 days, and were characterized by increased cellular influx in bronchoalveolar lavage fluid, interstitial pneumonia and gene expression changes. However, CNT(Large) elicited an earlier onset of inflammation and DNA damage, and induced more fibrosis and a unique fibrotic gene expression signature at day 28, compared to CNT(Small). The results indicate that the extent of change at the molecular level during early response phases following an acute exposure is greater in mice exposed to CNT(Large), which may eventually lead to the different responses observed at day 28.
authors.affiliation: National Research Centre for the Working Environment, Copenhagen DK-2100, Denmark; Department of Science, Systems and Models, Roskilde University, DK-4000 Roskilde, Denmark. Electronic address: spo@nrcwe.dk.
authors.forename: Sarah S
authors.initials: SS
authors.lastname: Poulsen
authors.affiliation: National Research Centre for the Working Environment, Copenhagen DK-2100, Denmark. Electronic address: ats@nrcwe.dk.
authors.forename: Anne T
authors.initials: AT
authors.lastname: Saber
authors.affiliation: Environmental and Radiation Health Sciences Directorate, Health Canada, Ottawa, Ontario K1A 0K9, Canada. Electronic address: Andrew.williams@hc-sc.gc.ca.
authors.forename: Andrew
authors.initials: A
authors.lastname: Williams
authors.affiliation: Department of Science, Systems and Models, Roskilde University, DK-4000 Roskilde, Denmark. Electronic address: oa@ruc.dk.
authors.forename: Ole
authors.initials: O
authors.lastname: Andersen
authors.affiliation: Department of Micro- and Nanotechnology, Technical University of Denmark, DK-2800 Kgs. Lyngby, Denmark. Electronic address: carko@nanotech.dtu.dk.
authors.forename: Carsten
authors.initials: C
authors.lastname: Købler
authors.affiliation: Nanologica AB, SE-114 28 Stockholm, Sweden. Electronic address: rba@nrcwe.dk.
authors.forename: Rambabu
authors.initials: R
authors.lastname: Atluri
authors.affiliation: Veneto Nanotech SCpA, ECSIN - European Centre for the Sustainable Impact of Nanotechnology, I-45100 Rovigo, Italy. Electronic address: mariaelena.pozzebon@yahoo.it.
authors.forename: Maria E
authors.initials: ME
authors.lastname: Pozzebon
authors.affiliation: Veneto Nanotech SCpA, ECSIN - European Centre for the Sustainable Impact of Nanotechnology, I-45100 Rovigo, Italy. Electronic address: stefano.pozzimucelli@venetonanotech.it.
authors.forename: Stefano P
authors.initials: SP
authors.lastname: Mucelli
authors.affiliation: Laboratory of Nanobiotechnology, National Institute for Research and Development in Microtechnologies, 077190 Bucharest, Romania. Electronic address: moni304ro@gmail.com.
authors.forename: Monica
authors.initials: M
authors.lastname: Simion
authors.affiliation: European Commission Joint Research Centre Institute for Environment and Sustainability, I-21027 Ispra, VA, Italy. Electronic address: david.rickerby@jrc.ec.europa.eu.
authors.forename: David
authors.initials: D
authors.lastname: Rickerby
authors.affiliation: National Food Institute, Technical University of Denmark, Søborg, Denmark. Electronic address: almo@food.dtu.dk.
authors.forename: Alicja
authors.initials: A
authors.lastname: Mortensen
authors.affiliation: National Research Centre for the Working Environment, Copenhagen DK-2100, Denmark. Electronic address: pja@nrcwe.dk.
authors.forename: Petra
authors.initials: P
authors.lastname: Jackson
authors.affiliation: National Research Centre for the Working Environment, Copenhagen DK-2100, Denmark. Electronic address: zky@nrcwe.dk.
authors.forename: Zdenka O
authors.initials: ZO
authors.lastname: Kyjovska
authors.affiliation: Department of Micro- and Nanotechnology, Technical University of Denmark, DK-2800 Kgs. Lyngby, Denmark. Electronic address: kristian.molhave@nanotech.dtu.dk.
authors.forename: Kristian
authors.initials: K
authors.lastname: Mølhave
authors.affiliation: National Research Centre for the Working Environment, Copenhagen DK-2100, Denmark. Electronic address: nrj@nrcwe.dk.
authors.forename: Nicklas R
authors.initials: NR
authors.lastname: Jacobsen
authors.affiliation: National Research Centre for the Working Environment, Copenhagen DK-2100, Denmark. Electronic address: kaj@nrcwe.dk.
authors.forename: Keld A
authors.initials: KA
authors.lastname: Jensen
authors.affiliation: Environmental and Radiation Health Sciences Directorate, Health Canada, Ottawa, Ontario K1A 0K9, Canada. Electronic address: Carole.yauk@hc-sc.gc.ca.
authors.forename: Carole L
authors.initials: CL
authors.lastname: Yauk
authors.affiliation: National Research Centre for the Working Environment, Copenhagen DK-2100, Denmark; Department of Public Health, University of Copenhagen, DK-1014 Copenhagen K, Denmark. Electronic address: hwa@nrcwe.dk.
authors.forename: Håkan
authors.initials: H
authors.lastname: Wallin
authors.affiliation: Environmental and Radiation Health Sciences Directorate, Health Canada, Ottawa, Ontario K1A 0K9, Canada. Electronic address: Sabina.halappanavar@hc-sc.gc.ca.
authors.forename: Sabina
authors.initials: S
authors.lastname: Halappanavar
authors.affiliation: National Research Centre for the Working Environment, Copenhagen DK-2100, Denmark; Department of Micro- and Nanotechnology, Technical University of Denmark, DK-2800 Kgs. Lyngby, Denmark. Electronic address: ubv@nrcwe.dk.
authors.forename: Ulla
authors.initials: U
authors.lastname: Vogel
mesh_terms: D000818:Animals; D001992:Bronchoalveolar Lavage Fluid / Q000166:cytology / Q000276:immunology; D004249:DNA Damage; D004305:Dose-Response Relationship, Drug; D005260:Female; D005786:Gene Expression Regulation; D053263:Gene Regulatory Networks; D018836:Inflammation Mediators / Q000378:metabolism*; D019570:Inhalation Exposure / Q000009:adverse effects; D008168:Lung / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D008810:Mice, Inbred C57BL; D037742:Nanotubes, Carbon / Q000633:toxicity*; D010316:Particle Size; D011014:Pneumonia / Q000139:chemically induced* / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D011658:Pulmonary Fibrosis / Q000139:chemically induced* / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D017382:Reactive Oxygen Species; D018570:Risk Assessment; D013499:Surface Properties; D013997:Time Factors; D043922:Toxicogenetics / Q000379:methods; D014158:Transcription, Genetic / Q000187:drug effects*
chemical_list: D018836:Inflammation Mediators; D037742:Nanotubes, Carbon; D017382:Reactive Oxygen Species
keywords: Acute phase response; DNA microarray; In vivo; Nanotoxicology; ROS production; Toxicogenomics