title: High-content imaging and gene expression analysis to study cell-nanomaterial interactions: the effect of surface hydrophobicity.
abstract: The effects of nanoparticle (NP)-related parameters on cellular interactions are currently uncertain as analysis is complicated by the combinatorial diversity arising from the array of size, shape and surface properties. Here, we present a validated multiparametric high-content imaging method, with the utility of this approach demonstrated by in-depth analysis of the role of hydrophobicity on the interaction of Au NPs with cultured cells. In this methodology, we evaluate cell viability, membrane damage, induction of reactive oxygen species, mitochondrial health, cell area, skewness and induction of autophagy. High-content cell cycle phase studies and in-depth gene expression studies then serve to elucidate the underlying mechanisms. The data reveal a clear influence of the degree of NP surface hydrophobicity with membrane damage and autophagy induction, which is stronger than the effect of surface charge, for charges ranging between -50 and +20 mV. All labeling experiments occur in the same format, and can be further supplemented with additional parameters providing a broadly accessible format for studying cell-NP interactions under highly reproducible conditions.
authors.affiliation: MoSAIC/Biomedical NMR Unit, Department of Medicine, Catholic University of Leuven, B3000 Leuven, Belgium.
authors.forename: Bella B
authors.initials: BB
authors.lastname: Manshian
authors.affiliation: Department of Chemistry, University of Massachusetts, 710 North Pleasant Street, Amherst, MA 01003, United States.
authors.forename: Daniel F
authors.initials: DF
authors.lastname: Moyano
authors.affiliation: Vlaams Instituut voor Biotechnologie (VIB), Center for the Biology of Disease, Leuven, Belgium; Center for Human Genetics and Leuven Institute for Neuroscience and Disease (LIND), University of Leuven (KU Leuven), Leuven, Belgium.
authors.forename: Nikky
authors.initials: N
authors.lastname: Corthout
authors.affiliation: Vlaams Instituut voor Biotechnologie (VIB), Center for the Biology of Disease, Leuven, Belgium; Center for Human Genetics and Leuven Institute for Neuroscience and Disease (LIND), University of Leuven (KU Leuven), Leuven, Belgium.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Munck
authors.affiliation: MoSAIC/Biomedical NMR Unit, Department of Medicine, Catholic University of Leuven, B3000 Leuven, Belgium.
authors.forename: Uwe
authors.initials: U
authors.lastname: Himmelreich
authors.affiliation: Department of Chemistry, University of Massachusetts, 710 North Pleasant Street, Amherst, MA 01003, United States. Electronic address: rotello@umass.chem.edu.
authors.forename: Vincent M
authors.initials: VM
authors.lastname: Rotello
authors.affiliation: MoSAIC/Biomedical NMR Unit, Department of Medicine, Catholic University of Leuven, B3000 Leuven, Belgium; Faculty of Pharmaceutical Sciences, Ghent University, Ghent, Belgium. Electronic address: Stefaan.Soenen@med.kuleuven.be.
authors.forename: Stefaan J
authors.initials: SJ
authors.lastname: Soenen
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D005786:Gene Expression Regulation / Q000187:drug effects; D006046:Gold / Q000737:chemistry / Q000378:metabolism* / Q000633:toxicity; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D051379:Mice; D053758:Nanoparticles / Q000737:chemistry / Q000378:metabolism* / Q000633:toxicity; D058953:Neural Stem Cells / Q000166:cytology / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D013499:Surface Properties
chemical_list: D017382:Reactive Oxygen Species; D006046:Gold
keywords: Cytotoxicity; High-content imaging; Nanoparticles; Nanotoxicology