title: Toxicity screenings of nanomaterials: challenges due to interference with assay processes and components of classic in vitro tests.
abstract: Given the multiplicity of nanoparticles (NPs), there is a requirement to develop screening strategies to evaluate their toxicity. Within the EU-funded FP7 NanoTEST project, a panel of medically relevant NPs has been used to develop alternative testing strategies of NPs used in medical diagnostics. As conventional toxicity tests cannot necessarily be directly applied to NPs in the same manner as for soluble chemicals and drugs, we determined the extent of interference of NPs with each assay process and components. In this study, we fully characterized the panel of NP suspensions used in this project (poly(lactic-co-glycolic acid)-polyethylene oxide [PLGA-PEO], TiO2, SiO2, and uncoated and oleic-acid coated Fe3O4) and showed that many NP characteristics (composition, size, coatings, and agglomeration) interfere with a range of in vitro cytotoxicity assays (WST-1, MTT, lactate dehydrogenase, neutral red, propidium iodide, (3)H-thymidine incorporation, and cell counting), pro-inflammatory response evaluation (ELISA for GM-CSF, IL-6, and IL-8), and oxidative stress detection (monoBromoBimane, dichlorofluorescein, and NO assays). Interferences were assay specific as well as NP specific. We propose how to integrate and avoid interference with testing systems as a first step of a screening strategy for biomedical NPs.
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 (RMCX) , 75205 Paris cedex 13 , France .
authors.forename: Rina
authors.initials: R
authors.lastname: Guadagnini
authors.forename: Blanka
authors.initials: B
authors.lastname: Halamoda Kenzaoui
authors.forename: Laura
authors.initials: L
authors.lastname: Walker
authors.forename: Giulio
authors.initials: G
authors.lastname: Pojana
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Magdolenova
authors.forename: Dagmar
authors.initials: D
authors.lastname: Bilanicova
authors.forename: Margaret
authors.initials: M
authors.lastname: Saunders
authors.forename: Lucienne
authors.initials: L
authors.lastname: Juillerat-Jeanneret
authors.forename: Antonio
authors.initials: A
authors.lastname: Marcomini
authors.forename: Anna
authors.initials: A
authors.lastname: Huk
authors.forename: Maria
authors.initials: M
authors.lastname: Dusinska
authors.forename: Lise M
authors.initials: LM
authors.lastname: Fjellsbø
authors.forename: Francelyne
authors.initials: F
authors.lastname: Marano
authors.forename: Sonja
authors.initials: S
authors.lastname: Boland
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D006801:Humans; D066298:In Vitro Techniques / Q000379:methods*; D053758:Nanoparticles / Q000633:toxicity*; D051381:Rats; D018675:Toxicity Tests / Q000379:methods*
keywords: NanoTEST; cytotoxicity; nanomedicine; nanoparticles; oxidative stress; pro-inflammatory response; testing strategy