Genetic toxicity assessment of engineered nanoparticles using a 3D in vitro skin model (EpiDerm ™ )

Wills, John W.; Hondow, Nicole; Thomas, Adam D.; Chapman, Katherine E.; Fish, David; Maffeis, Thierry G.; Penny, Mark W.; Brown, Richard A.; Jenkins, Gareth J.S.; Brown, Andy P.; White, Paul A.; Doak, Shareen H.

Abstract

© 2016 The Author(s). Background: The rapid production and incorporation of engineered nanomaterials into consumer products alongside research suggesting nanomaterials can cause cell death and DNA damage (genotoxicity) makes in vitro assays desirable for nanosafety screening. However, conflicting outcomes are often observed when in vitro and in vivo study results are compared, suggesting more physiologically representative in vitro models are required to minimise reliance on animal testing. Method: BASF Levasil® silica nanoparticles (16 and 85 nm) were used to adapt the 3D reconstructed skin micronucleus (RSMN) assay for nanomaterials administered topically or into the growth medium. 3D dose-responses were compared to a 2D micronucleus assay using monocultured human B cells (TK6) after standardising dose between 2D / 3D assays by total nanoparticle mass to cell number. Cryogenic vitrification, scanning electron microscopy and dynamic light scattering techniques were applied to characterise in-medium and air-liquid interface exposures. Advanced transmission electron microscopy imaging modes (high angle annular dark field) and X-ray spectrometry were used to define nanoparticle penetration / cellular uptake in the intact 3D models and 2D monocultured cells. Results: For all 2D exposures, significant (p

• Journal Article Type: Article
• Acceptance Date: Aug 30, 2016
• Online Publication Date: Sep 9, 2016
• Publication Date: Sep 9, 2016
• Deposit Date: Dec 14, 2017
• Publicly Available Date: Dec 14, 2017
• Journal: Particle and Fibre Toxicology
• Electronic ISSN: 1743-8977
• Publisher: BioMed Central
• Peer Reviewed: Peer Reviewed
• Volume: 13
• Article Number: 50
• DOI: https://doi.org/10.1186/s12989-016-0161-5
• Keywords: 3D cell culture, silica, genotoxicity, nanotoxicology, physico-chemical characterisation, nanoparticles, reconstructed skin, RSMN, micronucleus assay, air-liquid interface
• Public URL: https://uwe-repository.worktribe.com/output/908143
• Publisher URL: https://doi.org/10.1186/s12989-016-0161-5
• Contract Date: Dec 14, 2017