The Japanese Ring Study of a Human Cell Line Activation Test (h-CLAT) for Predicting Skin Sensitization Potential (6<sup>th</sup> Report): A Study for Evaluating Oxidative Hair Dye Sensitization Potential Using h-CLAT

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We are conducting a Japanese ring study to develop the human Cell Line Activation Test (h-CLAT). The aim of this study is to confirm whether the h-CLAT can predict skin sensitization for oxidative hair dyes. In addition, we studied the effect of test chemical fluorescence on prediction performance. h-CLATs were independently performed in two laboratories for eight chemicals, yielding good reproducibility between the laboratories. Good concordance was obtained between LLNA results and study chemicals. p-Phenylenediamine, which exhibits fluorescence at around 530 nm, which is in the h-CLAT measurement range, was correctly evaluated as positive by h-CLAT. It was possible to compensate for the influence of fluorescence in CD86/CD54 expression measurements, suggesting that substances exhiviting autofluorescence, such as oxidative hair dyes, can also be evaluated correctly. While a notably high, concentration-dependent CD54 expression was observed with Bandrowski's base, this expression was not observed with p-phenylenediamine, a result which suggests that the sensitization responses of these two dyes may differ. In conclusion, it is suggested that h-CLAT is useful for evaluating the skin sensitization potential of oxidative hair dyes.

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