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KMID : 1177720210150010015
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Journal of Alternatives to Animal Experiments
2021 Volume.15 No. 1 p.15 ~ p.22
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Establishment Study of ARE-Nrf2 Luciferase LuSens Skin Sensitization Test(OECD TG 442D) in Korea
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Hong Mi-Hye
Joe In-Suk
Bang Seo-Young
Yi Jung-Sun
Kim Kwang-Jin
Yoon Hae-Seong
Kim Tae-Sung
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Abstract
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This study aimed to establish the LuSens test method for identification of skin sensitisers in our laboratory and to facilitate the domestic use of the method. We utilized 10 recommended proficiency substances in OECD TG 442D consisting of 6 skin sensitisers (UN GHS category 1A and 1B: Eugenol, Cinnamyl alcohol, 2-Mercaptobenzo-thiazole, 4-Methylaminophenol sulfate, Methyl dibromo glutaronitrile and 2,4-Dinitro-chlorobenzene) and 4 non-sensitisers (No category: Salicylic acid, Glycerol, Isopropanol and Sulfanilamide). We measured the activity of luciferase induced by the test substances based on the CV75 that was determined by cytotoxicity dose-finding test. While the maximal luciferase fold induction values for each skin non-sensitisers ranged from 0.95 to 1.28, those for each skin sensitisers ranged from 1.96 to 5.66. We predicted sensitivity of the test substances on the basis of the luciferase fold induction values. Our results were within the range of acceptance criteria and accord with in vivo and in vitro references in OECD TG 442D. We found that the LuSens test method correctly identified proficiency substances into sensitisers and non-sensitisers. Therefore, the results obtained from the proficiency test demonstrated that we successfully introduced the LuSens test method in our laboratory. Furthermore, we have prepared a new in vitro skin sensitization test (ARE-Nrf2 luciferase LuSens) guideline for the safety evaluation of cosmetics and contributed to the dissemination of the method via technical transfer in Korea.
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KEYWORD
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skin sensitization, ARE-Nrf2 luciferase, LuSens, OECD TG 442D
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