Application of the KeratinoSens assay to prediction of dermal sensitization hazard for botanical cos

• 作 者: David GAN Kimberly NORMAN Nicole BARNES Hans A.RAABE Cristi GOMEZ Xuejun d.YIN John HARBELL
• 机 构: Mary Kay Inc.Dallas, TX IIVS, Gaithersburg, MD
• 出 处: 中国毒理学会第六届全国毒理学大会
• 发表日期: 2013年7期

　　OBJECTIVE The goal of the study was to determine the potential for the KeratinoSens assay to identify electrophile allergens within a botanical extract matrix.METHODS In the KeratinoSens assay, the induction of a luciferase gene under the control of the antioxidant response element (ARE) derived from the human gene AKR1C2 gene is quantified.In parallel, cytotoxicity is assessed by both Neutral Red Uptake (NRU) and MTT assays.Test samples included gluteraldehyde (GA), dimethyl maleate (DM) and cinnamic aldehyde (CA) spiked into four different botanical extracts (including different excipient solvent systems).Test concentrations ranged up to 1 g· L-1(of complete extract) and a test concentration was considered positive if the relative viability was ≥70% and the fold induction of luciferase was 1.5x relative to the solvent controls.Activity was measured relative to the EC1.5 of the neat sensitizer as a function of sensitizer concentration and extract composition.Three independent trials were performed on each test material.RESULTS No appreciable cytotoxicity was observed with any of the samples.Sensitizers spiked at 1% to 2% were sufficient to induce a positive up regulation of the ARE below the maximum test material concentration of up to 1 g· L-1.These responses occurred at cell survivals well in excess of 70%.The botanical matrix impacted recovery for both gluteraldehyde and dimethyl maleate sensitizers.The recovery of the GA spike required at least a-3 fold increase in concentration relative to the chemical alone and botanical extract #3 reduced the activity below detection.The DM and CA showed activity at about the same effective concentrations as the neat chemical although the DM showed reduced activity in botanical extract #3 as well.CONCLUSION These data suggest that the KeratinoSens assay has the potential to identify electrophile allergens within a botanical extract matrix.