title: Use of a high-throughput screening approach coupled with in vivo zebrafish embryo screening to develop hazard ranking for engineered nanomaterials.
abstract: Because of concerns about the safety of a growing number of engineered nanomaterials (ENM), it is necessary to develop high-throughput screening and in silico data transformation tools that can speed up in vitro hazard ranking. Here, we report the use of a multiparametric, automated screening assay that incorporates sublethal and lethal cellular injury responses to perform high-throughput analysis of a batch of commercial metal/metal oxide nanoparticles (NP) with the inclusion of a quantum dot (QD1). The responses chosen for tracking cellular injury through automated epifluorescence microscopy included ROS production, intracellular calcium flux, mitochondrial depolarization, and plasma membrane permeability. The z-score transformed high volume data set was used to construct heat maps for in vitro hazard ranking as well as showing the similarity patterns of NPs and response parameters through the use of self-organizing maps (SOM). Among the materials analyzed, QD1 and nano-ZnO showed the most prominent lethality, while Pt, Ag, SiO2, Al2O3, and Au triggered sublethal effects but without cytotoxicity. In order to compare the in vitro with the in vivo response outcomes in zebrafish embryos, NPs were used to assess their impact on mortality rate, hatching rate, cardiac rate, and morphological defects. While QDs, ZnO, and Ag induced morphological abnormalities or interfered in embryo hatching, Pt and Ag exerted inhibitory effects on cardiac rate. Ag toxicity in zebrafish differed from the in vitro results, which is congruent with this material's designation as extremely dangerous in the environment. Interestingly, while toxicity in the initially selected QD formulation was due to a solvent (toluene), supplementary testing of additional QDs selections yielded in vitro hazard profiling that reflect the release of chalcogenides. In conclusion, the use of a high-throughput screening, in silico data handling and zebrafish testing may constitute a paradigm for rapid and integrated ENM toxicological screening.
authors.affiliation: Department of Medicine, Division of NanoMedicine, Center for Environmental Implications of Nanotechnology, California NanoSystems Institute, Los Angeles, CA 90095, USA.
authors.forename: Saji
authors.initials: S
authors.lastname: George
authors.forename: Tian
authors.initials: T
authors.lastname: Xia
authors.forename: Robert
authors.initials: R
authors.lastname: Rallo
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhao
authors.forename: Zhaoxia
authors.initials: Z
authors.lastname: Ji
authors.forename: Sijie
authors.initials: S
authors.lastname: Lin
authors.forename: Xiang
authors.initials: X
authors.lastname: Wang
authors.forename: Haiyuan
authors.initials: H
authors.lastname: Zhang
authors.forename: Bryan
authors.initials: B
authors.lastname: France
authors.forename: David
authors.initials: D
authors.lastname: Schoenfeld
authors.forename: Robert
authors.initials: R
authors.lastname: Damoiseaux
authors.forename: Rong
authors.initials: R
authors.lastname: Liu
authors.forename: Shuo
authors.initials: S
authors.lastname: Lin
authors.forename: Kenneth A
authors.initials: KA
authors.lastname: Bradley
authors.forename: Yoram
authors.initials: Y
authors.lastname: Cohen
authors.forename: André E
authors.initials: AE
authors.lastname: Nel
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004625:Embryo, Nonmammalian / Q000187:drug effects* / Q000473:pathology; D008856:Microscopy, Fluorescence / Q000379:methods*; D049329:Nanostructures / Q000633:toxicity*; D045663:Quantum Dots; D018675:Toxicity Tests / Q000379:methods*; D015027:Zebrafish