title: High-throughput screening platform for engineered nanoparticle-mediated genotoxicity using CometChip technology.
abstract: The likelihood of intentional and unintentional engineered nanoparticle (ENP) exposure has dramatically increased due to the use of nanoenabled products. Indeed, ENPs have been incorporated in many useful products and have enhanced our way of life. However, there are many unanswered questions about the consequences of nanoparticle exposures, in particular, with regard to their potential to damage the genome and thus potentially promote cancer. In this study, we present a high-throughput screening assay based upon the recently developed CometChip technology, which enables evaluation of single-stranded DNA breaks, abasic sites, and alkali-sensitive sites in cells exposed to ENPs. The strategic microfabricated, 96-well design and automated processing improves efficiency, reduces processing time, and suppresses user bias in comparison to the standard comet assay. We evaluated the versatility of this assay by screening five industrially relevant ENP exposures (SiO2, ZnO, Fe2O3, Ag, and CeO2) on both suspension human lymphoblastoid (TK6) and adherent Chinese hamster ovary (H9T3) cell lines. MTT and CyQuant NF assays were employed to assess cellular viability and proliferation after ENP exposure. Exposure to ENPs at a dose range of 5, 10, and 20 μg/mL induced dose-dependent increases in DNA damage and cytotoxicity. Genotoxicity profiles of ZnO>Ag>Fe2O3>CeO2>SiO2 in TK6 cells at 4 h and Ag>Fe2O3>ZnO>CeO2>SiO2 in H9T3 cells at 24 h were observed. The presented CometChip platform enabled efficient and reliable measurement of ENP-mediated DNA damage, therefore demonstrating the efficacy of this powerful tool in nanogenotoxicity studies.
authors.affiliation: Department of Environmental Health, Center for Nanotechnology and Nanotoxicology, Harvard School of Public Health , Boston, Massachusetts 02115, United States.
authors.forename: Christa
authors.initials: C
authors.lastname: Watson
authors.forename: Jing
authors.initials: J
authors.lastname: Ge
authors.forename: Joel
authors.initials: J
authors.lastname: Cohen
authors.forename: Georgios
authors.initials: G
authors.lastname: Pyrgiotakis
authors.forename: Bevin P
authors.initials: BP
authors.lastname: Engelward
authors.forename: Philip
authors.initials: P
authors.lastname: Demokritou
mesh_terms: D000818:Animals; D016466:CHO Cells; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D020552:Comet Assay / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D053904:DNA Breaks, Single-Stranded / Q000187:drug effects; D004738:Engineering; D005290:Ferric Compounds / Q000737:chemistry / Q000633:toxicity; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D009153:Mutagens / Q000737:chemistry* / Q000633:toxicity*; D053758:Nanoparticles / Q000633:toxicity*; D012822:Silicon Dioxide / Q000737:chemistry / Q000633:toxicity; D012834:Silver / Q000737:chemistry / Q000633:toxicity; D013329:Structure-Activity Relationship; D013997:Time Factors
chemical_list: D005290:Ferric Compounds; D009153:Mutagens; C000499:ferric oxide; D012834:Silver; D012822:Silicon Dioxide