Abstract:
The rabbit skin irritation test has been the standard for evaluating the irritation potential of chemicals; however, alternative methods that do not use animal testing are actively encouraged. Reconstructed human epidermis (RhE) models mimic the biochemical and physiological properties of the human epidermis and can be used as an alternative method. On RhE methods, the metabolic activity of RhE models is used to predict skin irritation, with a reduction in metabolic activity indicating a reduced number of viable cells and linking cell death to skin irritation processes. However, new challenges have emerged as the use of RhE models increases, including the need for non-invasive and marker-free methodologies to assess cellular states. Electrochemical impedance spectroscopy (EIS) is one such methodology that can meet these requirements. In this study, our results showed that EIS can differentiate between irritant and non-irritant chemicals, with a significant increase in the capacitance values observed in the irritant samples. A ROC curve analysis showed that the prediction method based on EIS met OECD TG 439 requirements at all time points and had 95% within-laboratory reproducibility. Comparison with the MTT viability assay showed that prediction using EIS achieved higher sensitivity, specificity, and accuracy. These results suggest that EIS could potentially replace animal testing in the evaluation of irritation potential and could be a valuable addition to in vitro testing strategies.
摘要:
兔皮肤刺激试验已成为评估化学品潜在刺激的标准;然而,积极鼓励不使用动物试验的替代方法.重建的人表皮(RhE)模型模拟人表皮的生化和生理特性,并且可以用作替代方法。在RhE方法上,RhE模型的代谢活动用于预测皮肤刺激,代谢活性降低表明活细胞数量减少,并将细胞死亡与皮肤刺激过程联系起来。然而,随着RHE模型使用的增加,新的挑战出现了,包括需要非侵入性和无标记的方法来评估细胞状态。电化学阻抗谱(EIS)是可以满足这些要求的一种这样的方法。在这项研究中,我们的结果表明,EIS可以区分刺激性和非刺激性化学物质,在刺激性样品中观察到的电容值显着增加。ROC曲线分析表明,基于EIS的预测方法在所有时间点都满足OECDTG439的要求,并且在实验室内具有95%的可重复性。与MTT活力测定的比较表明,使用EIS进行预测具有更高的灵敏度,特异性,和准确性。这些结果表明,EIS可以在评估刺激潜力方面替代动物测试,并且可能是体外测试策略的有价值的补充。