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Development of screening assays for the assessment of the toxicity of migrants from commodities in keratinocytes
Project
Project code: BfR-CPS-02-7Z-003
Contract period: 01.01.2002
- 31.12.2006
Purpose of research: Applied research
Studies on toxic and sensitizing potential of chemically defined mixtures of substances and migrants from cosmetics and commodities in human keratinocytes (primary keratinocytes, HaCat cell line). The cytotoxic effects are characterized by assays for toxicity (neutral red assay) and markers for proliferation (Ki67- immunoreactivity) and apoptosis (with TUNEL and Hoechst 33342 staining).
For the detection of toxic effects in primary human keratinocytes and a human keratinocyte cell line (HaCaT), apoptosis and proliferation were studied simultaneously using a novel staining technique (TUNEL and Ki67- immunoreactivity) in addition to the determination of cytotoxicity (MTT, Neutral red assay). Pesticides were used as model compounds. Treatment with low concentrations of single pesticides resulted in moderate effects (0-10% cytotoxicity compared to controls), whereas binary mixtures of pesticides induced a pronounced increase in the number of apoptotic (TUNEL-positive) cells and a strong decrease in the number of proliferating (Ki67-positive) cells compared to controls. Induction of apoptosis and inhibition of proliferation were found to be more sensitive parameters than cytotoxicity for which higher concentrations of pesticides were needed to show significant effects. Thus, the former parameters might be useful for the detection of the toxicity of migrants from commodities or cosmetic articles.
Section overview
Subjects
- Toxicology
