title: Silica nanoparticles administered at the maximum tolerated dose induce genotoxic effects through an inflammatory reaction while gold nanoparticles do not.
abstract: While the collection of genotoxicity data and insights into potential mechanisms of action for nano-sized particulate materials (NPs) are steadily increasing, there is great uncertainty whether current standard assays are suitable to appropriately characterize potential risks. We investigated the effects of NPs in an in vivo Comet/micronucleus (MN) combination assay and in an in vitro MN assay performed with human blood. We also incorporated additional endpoints into the in vivo study in an effort to delineate primary from secondary mechanisms. Amorphous silica NPs (15 and 55 nm) were chosen for their known reactivity, while gold nano/microparticles (2, 20, and 200 nm) were selected for their wide size range and lower reactivity. DNA damage in liver, lung and blood cells and micronuclei in circulating reticulocytes were measured after 3 consecutive intravenous injections to male Wistar rats at 48, 24 and 4h before sacrifice. Gold nano/microparticles were negative for MN induction in vitro and in vivo, and for the induction of DNA damage in all tissues. Silica particles, however, caused a small but reproducible increase in DNA damage and micronucleated reticulocytes when tested at their maximum tolerated dose (MTD). No genotoxic effects were observed at lower doses, and the in vitro MN assay was also negative. We hypothesize that silica NPs initiate secondary genotoxic effects through release of inflammatory cell-derived oxidants, similar to that described for crystalline silica (quartz). Such a mechanism is supported by the occurrence of increased neutrophilic infiltration, necrosis, and apoptotic cells in the liver, and induction of inflammatory markers TNF-α and IL-6 in plasma at the MTDs. These results were fairly consistent between silica NPs and the quartz control, thereby strengthening the argument that silica NPs may act in a similar, thresholded manner. The observed profile is supportive of a secondary genotoxicity mechanism that is driven by inflammation.
authors.affiliation: The Procter and Gamble Co., Miami Valley Innovation Center, 11810 East Miami River Road, Cincinnati, OH 45252, USA.
authors.forename: Thomas R
authors.initials: TR
authors.lastname: Downs
authors.forename: Meredith E
authors.initials: ME
authors.lastname: Crosby
authors.forename: Ting
authors.initials: T
authors.lastname: Hu
authors.forename: Shyam
authors.initials: S
authors.lastname: Kumar
authors.forename: Ashley
authors.initials: A
authors.lastname: Sullivan
authors.forename: Katherine
authors.initials: K
authors.lastname: Sarlo
authors.forename: Bob
authors.initials: B
authors.lastname: Reeder
authors.forename: Matt
authors.initials: M
authors.lastname: Lynch
authors.forename: Matthew
authors.initials: M
authors.lastname: Wagner
authors.forename: Tim
authors.initials: T
authors.lastname: Mills
authors.forename: Stefan
authors.initials: S
authors.lastname: Pfuhler
mesh_terms: D000818:Animals; D020552:Comet Assay; D004249:DNA Damage; D004305:Dose-Response Relationship, Drug; D006046:Gold / Q000633:toxicity*; D006801:Humans; D007249:Inflammation / Q000139:chemically induced; D008099:Liver / Q000187:drug effects; D008168:Lung / Q000187:drug effects; D008297:Male; D020714:Maximum Tolerated Dose; D015162:Micronucleus Tests; D009153:Mutagens / Q000633:toxicity*; D049329:Nanostructures / Q000633:toxicity*; D010316:Particle Size; D011791:Quartz / Q000633:toxicity; D051381:Rats; D017208:Rats, Wistar; D012822:Silicon Dioxide / Q000633:toxicity*
chemical_list: D009153:Mutagens; D011791:Quartz; D006046:Gold; D012822:Silicon Dioxide