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Toxicological Evaluation of SiO2 Nanoparticles by Zebrafish Embryo Toxicity Test
Sandra Vranic 1,2,υ,φ, Yasuhito Shimada 3,4,υ , Sahoko Ichihara 5,* , Masayuki Kimata 6, Wenting Wu 1,, Toshio Tanaka 7, Sonja Boland 8 , Lang Tran 9 and Gaku Ichihara 1,2
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* Correspondence: saho@jichi.ac.jp; Tel.: +81-285-58-7335; Fax: +81-285-44-8645
υ These authors contributed equally to this work.
φ Current Affiliation: Nanomedicine Lab, Faculty of Biology, Medicine and Health,
The University of Manchester, AV Hill Building, Manchester M13 9PT, UK. Received: 30 December 2018; Accepted: 10 February 2019; Published: 18 February 2019
Department of Occupational and Environmental Health, Nagoya University Graduate School of Medicine, Nagoya 466-8560, Japan; sandra.vranic@manchester.ac.uk (S.V.); wendywu1206@163.com (W.W.); gak@rs.tus.ac.jp (G.I.)
Department of Occupational and Environmental Medicine, Faculty of Pharmaceutical Sciences,
Tokyo University of Sciences, Noda 278-8510, Japan
Department of Integrative Pharmacology, Mie University Graduate School of Medicine, Tsu 514-8572, Japan; shimada.yasuhito@mie-u.ac.jp
Mie University Zebrafish Drug Screening Center, Tsu 514-8572, Japan
Department of Environmental and Preventive Medicine, Jichi Medical University School of Medicine, Shimotsuke 329-0498, Japan
Department of Human Functional Genomics, Life Science Research Center, Mie University,
Tsu 514-8572, Japan; mk.lucky-seven@ezweb.ne.jp
Department of Systems Pharmacology, Mie University Graduate School of Medicine, Tsu 514-8572, Japan; tanaka@doc.medic.mie-u.ac.jp
Unit of Functional and Adaptive Biology (BFA), Laboratory of Molecular and Cellular Responses to Xenobiotics, CNRS UMR 8251, Universite Paris Diderot, Sorbonne Paris Cite, 75013 Paris, France; boland@univ-paris-diderot.fr
Institute of Occupational Medicine, Research Avenue North, Riccarton, Edinburgh EH14 4AP, UK; lang.tran@iom-world.org
Abstract: As the use of nanoparticles (NPs) is increasing, the potential toxicity and behavior of NPs in living systems need to be better understood. Our goal was to evaluate the developmental toxicity and bio-distribution of two different sizes of fluorescently-labeled SiO2 NPs, 25 and 115 nm, with neutral surface charge or with different surface functionalization, rendering them positively or negatively charged, in order to predict the effect of NPs in humans. We performed a zebrafish embryo toxicity test (ZFET) by exposing the embryos to SiO2 NPs starting from six hours post fertilization (hpf). Survival rate, hatching time, and gross morphological changes were assessed at 12, 24, 36, 48, 60, and 72 hpf. We evaluated the effect of NPs on angiogenesis by counting the number of sub-intestinal vessels between the second and seventh intersegmental vessels and gene expression analysis of vascular endothelial growth factor (VEGF) and VEGF receptors at 72 hpf. SiO2 NPs did not show any adverse effects on survival rate, hatching time, gross morphology, or physiological angiogenesis. We found that SiO2 NPs were trapped by the chorion up until to the hatching stage. After chemical removal of the chorion (dechorionation), positively surface-charged SiO2 NPs (25 nm) significantly reduced the survival rate of the fish compared to the control group. These results indicate that zebrafish chorion acts as a physical barrier against SiO2 NPs, and removing the chorions in ZFET might be necessary for evaluation of toxicity of NPs.
Int. J. Mol. Sci. 2019, 20, 882; doi:10.3390/ijms20040882 www.mdpi.com/journal/ijms
Department of Systems Pharmacology Mie University Graduate School of Medicine
Mie University Medical Zebrafish Research Center