The Japanese Ring Study of a Human Cell Line Activation Test (h-CLAT) for Predicting Skin Sensitization Potential (5th Report): A Study for Evaluating Preservative Skin Sensitization Potential Using h-CLAT

Nanae Kosaka1, Hiroyuki Inaba2, Kenji Okamoto3, Makoto Mizuno4, Sakiko Sono5, Yoshinao Kato6, Masataka Kishi6, Takao Ashikaga5, Yuko Okamoto4, Hirofumi Kuwahara3, Tsuneaki Nakamura2, Hitoshi Sakaguchi1 and Yasuo Ohno7

1Kao Corporation, Tochigi, Japan; 2Lion Corporation, Kanagawa, Japan; 3Kanebo Cosmetics Inc., Kanagawa, Japan; 4 KOSÉ Corporation, Tokyo, Japan; 5Shiseido Co., Ltd., Kanagawa, Japan; 6Nippon Menard Cosmetic Co., Ltd., Aichi, Japan; 7National Institute of Health Science, Tokyo, Japan

AATEX 15(2):71-80, 2010

We have developed the human Cell Line Activation Test (h-CLAT) as an in vitro skin sensitization test. In this study, in order to examine whether h-CLAT can predict the skin sensitizing potential of a variety of preservatives, the ring study was performed in two independent laboratories. We selected a total of 16 preservatives, which have been used in cosmetic products at one time, and two different solvents, physiological saline (saline) and dimethyl sulfoxide (DMSO). According to h-CLAT protocol, expression of CD86 and CD54 on THP-1 cells was measured by flow cytometry after 24 h treatment with each chemical at 8 doses. The skin sensitizing potential for 13 of 16 preservatives was correctly predicted in one laboratory and 12 of 16 preservatives in the other. These data indicate relatively high concordance with the animal tests. Furthermore, five preservatives with sensitizing potential were tested using both solvents to compare the effect of saline and DMSO on the predicting capacity of h-CLAT. The sensitizing potential of these preservatives was correctly identified in both saline and DMSO. Our results from this smallscale ring study demonstrate the utility of h-CLAT for detecting the preservatives with skin sensitizing potential.

key words: h-CLAT, skin sensitization, alternatives, THP-1, preservative


(AATEX: Altern. Animal Test. EXperiment.: Alternatives to Animal Testing and EXperimentation)