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Development of fibrin hydrogel–based in vitro bioassay system for assessment of skin permeability to and pro-inflammatory activity mediated by zinc ion released from nanoparticles

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Abstract

Nanoparticles (NPs) are promising products in industry and medicine due to their unique physicochemical properties. In particular, zinc oxide (ZnO) NPs are extensively incorporated into sunscreens to protect the skin from exposure to ultraviolet radiation. However, there are several health concerns about skin penetration and the resultant toxicity. As methodologies for evaluating NP toxicity are under development, it is difficult to fully assess the toxicity of ZnO NPs toward humans. In this study, we developed a platform to simultaneously detect skin permeability to and pro-inflammatory activity mediated by zinc ion released from NPs. First, we generated a stable reporter cell line expressing green fluorescent protein (GFP) under the control of interleukin-8 (IL-8) promoter activity. The expression levels of GFP induced by zinc reflected the endogenous IL-8 expression levels and the pro-inflammatory responses. Next, we found that fibrin hydrogel can reproduce permeability to zinc ion of a human skin equivalent model and is therefore a promising material to assess skin permeability to zinc ion. Then, we constructed a fibrin hydrogel–based in vitro bioassay system for the simultaneous detection of skin permeability to and pro-inflammatory activity mediated by zinc ion released from NPs by using a stable reporter cell line and a fibrin hydrogel layer. This bioassay system is a promising in vitro permeation test due to its technical simplicity and good predictability. Overall, we believe that our bioassay system can be widely used in the cosmetics and pharmaceutical industries.

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Abbreviations

3D:

Three-dimensional

DMEM:

Dulbecco’s modified Eagle’s medium

ELISA:

Enzyme-linked immunosorbent assay

FBS:

Fetal bovine serum

GFP:

Green fluorescent protein

HSEM:

Human skin equivalent model

ICP-OES:

Inductively coupled plasma optical emission spectrometer

IL-8:

Interleukin-8

LDH:

Lactate dehydrogenase

NF-κB:

Nuclear factor kappa B

NP:

Nanoparticle

OECD:

Organisation for Economic Cooperation and Development

PBS:

Phosphate-buffered saline

SC:

Stratum corneum

SCCS:

Scientific Committee on Consumer Safety

TiO2 :

Titanium dioxide

TNF-α:

Tumor necrosis factor alpha

UV:

Ultraviolet

ZnO:

Zinc oxide

ZnO-Si:

Silica-coated zinc oxide

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Acknowledgments

We thank S. Sugino and A. Tada of AIST for excellent technical assistance.

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Author notes

  1. Yosuke Tabei and Waka Lin contributed equally to this work.

Authors and Affiliations

  1. Health and Medical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 2217-14 Hayashi-cho, Takamatsu, Kagawa, 761-0395, Japan

    Yosuke Tabei, Akinari Sonoda & Masanori Horie

  2. Biomedical Business Center, Healthcare Business Group, Ricoh Company, Ltd., 3-25-22 Tonomachi, Kawasaki-ku, Kawasaki, Kanagawa, 210-0821, Japan

    Waka Lin, Shusaku Shiomoto & Tomoaki Nakayama

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  1. Yosuke Tabei
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  2. Waka Lin
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Correspondence to Yosuke Tabei.

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Conflict of interest

W. Lin, S. Shiomoto, and T. Nakayama are employees of the sponsor of this study. However, this did not influence the objectivity of the study. The authors declare that they have no conflicts of interest.

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Tabei, Y., Lin, W., Shiomoto, S. et al. Development of fibrin hydrogel–based in vitro bioassay system for assessment of skin permeability to and pro-inflammatory activity mediated by zinc ion released from nanoparticles. Anal Bioanal Chem 412, 8269–8282 (2020). https://doi.org/10.1007/s00216-020-02970-5

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