[Determination of the irritating potential of environmental substances in an epidermal cell suspension in vitro].

B Wilke, I Hoth, B Bandemir
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Abstract

A simple in vitro screening method was established, which allows estimation of the effect of environmental products on cell alteration or irritative potential. Primary cell suspensions, prepared from the epidermis of guinea pig ears through trypsination (0.2%) over night at 4 degrees C, were used. The cell suspension (3 mill./ml) and test substances were mixed in equal portions (500 microliters) and incubated at room temperature for approximately 90 min. The degree of cell alteration was determined by the trypan blue exclusion test. Chemically defined substances as well as disinfectants were employed. The cell alteration was clearly dependent upon substance concentration as well as incubation time. On the basis of the mean alteration time (AT50) measured in vitro, and the mean irritation time (IT50) determined in the patch test in vivo, the substances were ranked according to strength of cell alteration or irritative potency in vitro and in vivo. A good correlation (Spearman's rank correlation, rs = 0.76) was found between the in vitro and in vivo results. Thus, this method is suited for the screening of chemical products especially in industrial laboratories.

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[体外测定表皮细胞悬浮液中环境物质的刺激电位]。
建立了一种简单的体外筛选方法,可以估计环境产物对细胞改变或刺激电位的影响。使用豚鼠耳表皮经胰蛋白酶作用(0.2%)在4℃下过夜制备的原代细胞悬液。将细胞悬液(3 ml ./ml)与试验物质按等量(500微升)混合,室温孵育约90分钟。通过台盼蓝排斥试验测定细胞改变程度。使用了化学定义的物质和消毒剂。细胞变化明显依赖于物质浓度和孵育时间。根据体外测得的平均改变时间(AT50)和体内斑贴试验测定的平均刺激时间(IT50),根据细胞改变强度或体外和体内的刺激效力对物质进行排序。体外实验结果与体内实验结果具有良好的相关性(Spearman秩相关,rs = 0.76)。因此,这种方法适用于化学产品的筛选,特别是在工业实验室。
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