title: Impact of serum as a dispersion agent for in vitro and in vivo toxicological assessments of TiO2nanoparticles.
abstract: Nanoparticles (NP) have a tendency to agglomerate after dispersion in physiological media, which can be prevented by the addition of serum. This may however result in modification of the toxic potential of particles due to the formation of protein corona. Our study aimed to analyze the role of serum that is added to improve the dispersion of 10 nm TiO2NPs on in vitro and in vivo effects following the exposure via the respiratory route. We characterized NP size, surface charge, sedimentation rate, the presence of protein corona and the oxidant-generating capacity after NP dispersion in the presence/absence of serum. The effect of serum on NP internalization, cytotoxicity and pro-inflammatory responses was assessed in a human pulmonary cell line, NCI-H292. Serum in the dispersion medium led to a slower sedimentation, but an enhanced cellular uptake of TiO2NPs. Despite this greater uptake, the pro-inflammatory response in NCI-H292 cells was lower after serum supplementation (used either as a dispersant or as a cell culture additive), which may be due to a reduced intrinsic oxidative potential of TiO2NPs. Interestingly, serum could be added 2 h after the NP treatment without affecting the pro-inflammatory response. We also determined the acute pulmonary and hepatic toxicity in vivo 24 h after intratracheal instillation of TiO2NPs in C57BL/6N mice. The use of serum resulted in an underestimation of the local acute inflammatory response in the lung, while a systemic response on glutathione reduction remained unaffected. In conclusion, serum as a dispersion agent for TiO2NPs can lead to an underestimation of the acute pro-inflammatory response in vitro and in vivo. To avoid potential unwanted effects of dispersants and medium components, we recommend that the protocol of NM preparation should be thoroughly tested, and reflect as close as possible realistic exposure conditions.
authors.affiliation: Univ Paris Diderot (Sorbonne Paris Cité), UMR 8251 CNRS, Unit of Functional and Adaptive Biology (BFA), Laboratory of Molecular and Cellular Responses to Xenobiotics, Univ Paris Diderot, 5 rue Thomas Mann, 75205, Paris cedex 13, France.|Nanomedicine Lab, Faculty of Medical and Human Sciences, University of Manchester, AV Hill Building, Upper Brook Street, Manchester, M13 9PT, UK.
authors.forename: Sandra
authors.initials: S
authors.lastname: Vranic
authors.affiliation: Centre for Sustainability, Environment and Health, National Institute for Public Health and the Environment, Bilthoven, The Netherlands.
authors.forename: Ilse
authors.initials: I
authors.lastname: Gosens
authors.affiliation: Danish Centre for Nanosafety, National Research Centre for the Working Environment, Copenhagen, Denmark.
authors.forename: Nicklas Raun
authors.initials: NR
authors.lastname: Jacobsen
authors.affiliation: Danish Centre for Nanosafety, National Research Centre for the Working Environment, Copenhagen, Denmark.
authors.forename: Keld A
authors.initials: KA
authors.lastname: Jensen
authors.affiliation: Centre for Safety of Substances and Products, National Institute for Public Health and the Environment, Bilthoven, The Netherlands.
authors.forename: Bas
authors.initials: B
authors.lastname: Bokkers
authors.affiliation: School of Life Sciences, Heriot-Watt University, John Muir building, Edinburgh, UK.|Department of Public Health, Section of Environmental Health, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Ali
authors.initials: A
authors.lastname: Kermanizadeh
authors.affiliation: School of Life Sciences, Heriot-Watt University, John Muir building, Edinburgh, UK.
authors.forename: Vicki
authors.initials: V
authors.lastname: Stone
authors.affiliation: Univ Paris Diderot (Sorbonne Paris Cité), UMR 8251 CNRS, Unit of Functional and Adaptive Biology (BFA), Laboratory of Molecular and Cellular Responses to Xenobiotics, Univ Paris Diderot, 5 rue Thomas Mann, 75205, Paris cedex 13, France.
authors.forename: Armelle
authors.initials: A
authors.lastname: Baeza-Squiban
authors.affiliation: Centre for Sustainability, Environment and Health, National Institute for Public Health and the Environment, Bilthoven, The Netherlands.
authors.forename: Flemming R
authors.initials: FR
authors.lastname: Cassee
authors.affiliation: Institute of Occupational Medicine, Edinburgh, UK.
authors.forename: Lang
authors.initials: L
authors.lastname: Tran
authors.affiliation: Univ Paris Diderot (Sorbonne Paris Cité), UMR 8251 CNRS, Unit of Functional and Adaptive Biology (BFA), Laboratory of Molecular and Cellular Responses to Xenobiotics, Univ Paris Diderot, 5 rue Thomas Mann, 75205, Paris cedex 13, France. boland@univ-paris-diderot.fr.
authors.forename: Sonja
authors.initials: S
authors.lastname: Boland
mesh_terms: D065567:Absorption, Physiological; D000280:Administration, Inhalation; D000818:Animals; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry / Q000276:immunology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D055598:Chemical Phenomena; D005260:Female; D008099:Liver / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D053768:Metal Nanoparticles / Q000008:administration & dosage / Q000737:chemistry / Q000633:toxicity* / Q000648:ultrastructure; D008810:Mice, Inbred C57BL; D016877:Oxidants / Q000008:administration & dosage / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D018384:Oxidative Stress / Q000187:drug effects; D010316:Particle Size; D014677:Pharmaceutical Vehicles / Q000737:chemistry*; D011897:Random Allocation; D020545:Respiratory Mucosa / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D044967:Serum / Q000737:chemistry*; D013499:Surface Properties; D013535:Suspensions; D014025:Titanium / Q000008:administration & dosage / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D023382:Toxicity Tests, Acute
chemical_list: D016877:Oxidants; D014677:Pharmaceutical Vehicles; D013535:Suspensions; C009495:titanium dioxide; D014025:Titanium
keywords: C57BL/6N; Inflammation; NCI-H292; Nanomaterial; Pulmonary toxicity; Titanium