FIRST PHASE INTER-LABORATORY VALIDATION OF THE IN VITRO EYE IRRITATION TESTS FOR COSMETIC INGREDIENTS: (8) EVALUATION OF CYTOTOXICITY TESTS ON SIRC CELLS

HIROSHI ITAGAKI1,2, MICHIO SHIBATA1,2, NAOKO TANI1,3, SHIGEMI KINOSHITA1,3, HIORSHI KAKASHIMA1,4, YOSHIAKI SEYAMA1,4, JUNKO OHUCHI1,5 YUTAKA KASAI1,5, JOSHIN OKADA1,5, HAJIME KOJIMA1,6, YUUKO OKAMOTO1,7, MAYUMI KOTANI1,8, YASUO OHNO9, ATSUKO MIYAJIMA9 and AKIRA TAKANAKA1,2
1Japan Cosmetic Industry Assoc. (JCIA), 4th floor Hatsumei Bldg., 9-14, Tranomon, 2-chome, Minato-ku, Tokyo 105, Japan; 2Shiseido Safety & Analytical Research Center, 1050 Nippa-cho, Kohoku-ku. Yokohama 223, Japan; 3POLA Corporation, 560 Kashio-cho, Totsuka-ku, Yokohama 244, Japan; 4Kanebo Cosmetic Laboratory, 3-28, 5-chome, Kotobuki-cho, Odawara-shi, Kanagawa 256, Japan; 5KAO Corp. Tochigi Biological Sciences, 2606 Akabane, Ichikai, Haga, Tochigi 321-34, Japan; 6Biochemical Research Institute, Nippon Menard Cosmetic Co Ltd., 4-66, Asakusa, Ohgaki-shi, Gifu 503, Japan; 7Div. Fundamental Research, KOSE Corporation, 1-18-4 Azusawa, Itabushi-ku, Tokyo 174, Japan; 8Div. of Phamacology, Biological Safety Resource Center (BSRC), Mitamura Building, 26-1 Kaitaicho, Shinjuku-ku, Tokyo 112-0012 Japan 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158, Japan

Regular Article :AATEX 3(4):182-190

Abstract
Cytotoxicity tests using SIRC cells and crystal violet (CV) staining or neutral red (NR) uptake as their endpoints were primarily validated using nine surfactants and physiological saline by six and seven laboratories. respectively, under the Ministry of Health and Welfare (MHW) project on alternatives, to the Draize rabbit eye irritation test (Draize test) for cosmetic ingredients. The inter-laboratory coefficient of variation of EC50s of the test substances by CV staining and the NR uptake method were 0.262 and 0.304, respectively. These two endpoints gave similar EC5050s and the correlation coefficient between them was high (r=0.996) Correlation coefficients between maximal average Draize total scores (MAS) of test substances in Draize tests and their EC50s in CV staining and NR uptake were -0.894 and -0.913, respectively.
From these results, we concluded these cytotoxicity tests using SIRC cells are promising as alternative methods to the Draize test for cosmetic ingredients, and the CV staining and NR uptake methods gave almost identical results. However, further validation of these methods is needed using a wider range of cosmetic ingredients.

Key words: validation study, Draize eye irritation test, alternative, in vitro, cytotoxicity test, SIRC cell line, crystal violet, neutral red, surfactant.


return