title: Genotoxic and inflammatory effects of nanofibrillated cellulose in murine lungs.
abstract: Nanofibrillated cellulose (NFC) is a sustainable and renewable nanomaterial, with diverse potential applications in the paper and medical industries. As NFC consists of long fibres of high aspect ratio, we examined here whether TEMPO-(2,2,6,6-tetramethyl-piperidin-1-oxyl) oxidised NFC (length 300-1000nm, thickness 10-25nm), administrated by a single pharyngeal aspiration, could be genotoxic to mice, locally in the lungs or systemically in the bone marrow. Female C57Bl/6 mice were treated with four different doses of NFC (10, 40, 80 and 200 µg/mouse), and samples were collected 24h later. DNA damage was assessed by the comet assay in bronchoalveolar lavage (BAL) and lung cells, and chromosome damage by the bone marrow erythrocyte micronucleus assay. Inflammation was evaluated by BAL cell counts and analysis of cytokines and histopathological alterations in the lungs. A significant induction of DNA damage was observed at the two lower doses of NFC in lung cells, whereas no increase was seen in BAL cells. No effect was detected in the bone marrow micronucleus assay, either. NFC increased the recruitment of inflammatory cells to the lungs, together with a dose-dependent increase in mRNA expression of tumour necrosis factor α, interleukins 1β and 6, and chemokine (C-X-C motif) ligand 5, although there was no effect on the levels of the respective proteins. The histological analysis showed a dose-related accumulation of NFC in the bronchi, the alveoli and some in the cytoplasm of macrophages. In addition, neutrophilic accumulation in the alveolar lung space was observed with increasing dose. Our findings showed that NFC administered by pharyngeal aspiration caused an acute inflammatory response and DNA damage in the lungs, but no systemic genotoxic effect in the bone marrow. The present experimental design did not, however, allow us to determine whether the responses were transient or could persist for a longer time.
authors.affiliation: Finnish Institute of Occupational Health, P.O. Box 40, FI-00251 Helsinki, Finland, julia.catalan@ttl.fi.|Department of Anatomy, Embryology and Genetics, University of Zaragoza, Miguel Servet 177, 50.013 Zaragoza, Spain.
authors.forename: Julia
authors.initials: J
authors.lastname: Catalán
authors.affiliation: Finnish Institute of Occupational Health, P.O. Box 40, FI-00251 Helsinki, Finland.
authors.forename: Elina
authors.initials: E
authors.lastname: Rydman
authors.affiliation: Finnish Institute of Occupational Health, P.O. Box 40, FI-00251 Helsinki, Finland.
authors.forename: Kukka
authors.initials: K
authors.lastname: Aimonen
authors.affiliation: Finnish Institute of Occupational Health, P.O. Box 40, FI-00251 Helsinki, Finland.
authors.forename: Kati-Susanna
authors.initials: KS
authors.lastname: Hannukainen
authors.affiliation: Finnish Institute of Occupational Health, P.O. Box 40, FI-00251 Helsinki, Finland.
authors.forename: Satu
authors.initials: S
authors.lastname: Suhonen
authors.affiliation: Finnish Institute of Occupational Health, P.O. Box 40, FI-00251 Helsinki, Finland.
authors.forename: Esa
authors.initials: E
authors.lastname: Vanhala
authors.affiliation: Department of Anatomy, Embryology and Genetics, University of Zaragoza, Miguel Servet 177, 50.013 Zaragoza, Spain.
authors.forename: Carlos
authors.initials: C
authors.lastname: Moreno
authors.affiliation: Centre Technique du Papier, CS90251, 38044 Grenoble Cedex 9, France.
authors.forename: Valérie
authors.initials: V
authors.lastname: Meyer
authors.affiliation: Institut Technologique FCBA, CS90251, 38044 Grenoble Cedex 9, France.
authors.forename: Denilson da Silva
authors.initials: DD
authors.lastname: Perez
authors.affiliation: VTT Technical Research Centre of Finland Ltd, P.O. Box 1000, FI-02044 VTT, Finland.
authors.forename: Asko
authors.initials: A
authors.lastname: Sneck
authors.affiliation: VTT Technical Research Centre of Finland Ltd, P.O. Box 1000, FI-02044 VTT, Finland.
authors.forename: Ulla
authors.initials: U
authors.lastname: Forsström
authors.affiliation: Department of Biology, Linderstrøm-Lang Center for Protein Science, University of Copenhagen, Ole Maaloes Vej 5, DK-2200 Copenhagen N, Denmark and.
authors.forename: Casper
authors.initials: C
authors.lastname: Højgaard
authors.affiliation: Department of Biology, Linderstrøm-Lang Center for Protein Science, University of Copenhagen, Ole Maaloes Vej 5, DK-2200 Copenhagen N, Denmark and.
authors.forename: Martin
authors.initials: M
authors.lastname: Willemoes
authors.affiliation: Department of Biology, Linderstrøm-Lang Center for Protein Science, University of Copenhagen, Ole Maaloes Vej 5, DK-2200 Copenhagen N, Denmark and.
authors.forename: Jacob R
authors.initials: JR
authors.lastname: Winther
authors.affiliation: National Research Centre for the Working Environment, Lerso Parkallé 105, DK-2100 Copenhagen O, Denmark.
authors.forename: Ulla
authors.initials: U
authors.lastname: Vogel
authors.affiliation: Finnish Institute of Occupational Health, P.O. Box 40, FI-00251 Helsinki, Finland.
authors.forename: Henrik
authors.initials: H
authors.lastname: Wolff
authors.affiliation: Finnish Institute of Occupational Health, P.O. Box 40, FI-00251 Helsinki, Finland.
authors.forename: Harri
authors.initials: H
authors.lastname: Alenius
authors.affiliation: Finnish Institute of Occupational Health, P.O. Box 40, FI-00251 Helsinki, Finland.
authors.forename: Kai M
authors.initials: KM
authors.lastname: Savolainen
authors.affiliation: Finnish Institute of Occupational Health, P.O. Box 40, FI-00251 Helsinki, Finland.
authors.forename: Hannu
authors.initials: H
authors.lastname: Norppa
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000187:drug effects* / Q000378:metabolism; D002482:Cellulose / Q000494:pharmacology / Q000633:toxicity*; D020552:Comet Assay; D016207:Cytokines; D004247:DNA / Q000187:drug effects; D004249:DNA Damage; D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism; D005260:Female; D007249:Inflammation; D008168:Lung / Q000187:drug effects* / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects; D051379:Mice; D048629:Micronuclei, Chromosome-Defective; D015162:Micronucleus Tests; D057139:Nanofibers / Q000737:chemistry / Q000633:toxicity*
chemical_list: D016207:Cytokines; D002482:Cellulose; D004247:DNA