体外人源细胞激活试验法应用于4种染发剂的皮肤变态反应研究

Study of the Application of in Vitro Human Cell Line Activation Test with Four Hair Dyes

目的研究体外人源细胞激活试验h-CLAT法应用在染发剂的皮肤变态反应检测中的可行性,通过和现行化妆品标准中体内皮肤变态反应法的比较,评价体外人源细胞激活试验应用在染发剂皮肤变态反应试验上的优劣。方法参考OECD指导原则442E的体外人源细胞系激活试验方法,应用流式细胞仪技术在类树突样的人源白血病细胞系THP-1上对4种染发剂进行体外皮肤变态反应检查,并与体内皮肤变态反应法的检测结果相比较。结果体外h-CLAT试验表明,4种染发剂中#2,#3和#4染发剂的CD54RFI都<200且CD86RFI<150,即体外试验结果为阴性与体内皮肤变态反应试验比较,结果一致;而#1染发剂为体外结果阳性,且有效致敏浓度为315μg·m L-1,与体内变态反应阴性结果不同。结论与体内皮肤变态反应试验相比,体外人源细胞系激活试验在对染发剂的检验上有周期短、测试方法简便、评价结果客观等优点。缺点是无法反映整个过敏反应的过程,存在假阳性的可能。

OBJECTIVE To discuss the possibility of applying human cell line activation test in vitro to skin sensitization test for hair dyes. Compared with skin sensitization test in vivo, it was evaluated that advantages and disadvantages to apply human cell line activation test in vitro to skin sensitization test with hair dyes. METHODS In terms of OECD guideline 442 E, 4 different shampoos were tested in human cell line activation test with human leukemia THP-1 cells by flow cytometer. And this result was compared with result from Buehler test in vivo. RESULTS From human cell line activation test in vitro, it showed that #2, #3 and #4 hair dyes got the same negative results, from which showed all the CD54 RFI were 〈 200 and CD86 RFI were 〈 150, compared with the traditional skin sensitization test in vivo. However, #1 hair dye got the positive result from human cell line activation test, which were different from the result in Buehler test in vivo. CONCLUSION It has advantages of short test cycle, simple test method and objective evaluation in human cell line activation test in vitro for skin sensitization test with hair dyes, and disadvantages of reflecting part of process of skin sensitization, existing the possibility of false positive.