Reconstructed human cornea-like epithelium (RhCE) holds unprecedented promise for toxicological analyses and the replacement of animal use. However, current standards to evaluate potential ocular irritancy present a major downfall, the need to invasively alter tissue samples to evaluate cell viability. In this study, the applicability of impedance analysis was validated by monitoring the change in cell capacitance during tissue maturation and before and after chemical application using coupled electrodes. Our results indicate that cell maturation on RhCE models can be evaluated during model production using capacitance sensing offering a faster and simpler quality control criteria for RhCE model usability. Additionally, cell capacitance resulted to be more sensitive in detecting slight cell damages than methods based on cell metabolism, and when integrated into OECD-approved testing strategies, capacitance sensing performed as good as currently accepted methodologies displaying 66% sensitivity, 100% specificity and 83% accuracy when evaluated at 300 Hz. In summary, a quantitative analysis to predict in vivo ocular irritation based on changes in RhCE capacitance by impedance spectroscopy is suggested. This methodology represents a non-invasive and non-destructive alternative that would enable the monitoring of reversible effects or repeated dose toxicity.

重建的人类角膜样上皮 (RhCE) 为毒理学分析和替代动物使用带来了前所未有的希望。然而，目前评估潜在眼部刺激性的标准存在重大缺陷，即需要侵入性地改变组织样本以评估细胞活力。在这项研究中，通过使用耦合电极监测组织成熟期间和化学应用前后细胞电容的变化来验证阻抗分析的适用性。我们的结果表明，在模型生产过程中，可以使用电容传感评估 RhCE 模型上的细胞成熟度，从而为 RhCE 模型的可用性提供更快、更简单的质量控制标准。此外，细胞电容在检测轻微细胞损伤方面比基于细胞代谢的方法更敏感，当集成到经合组织批准的测试策略中时，电容传感的性能与目前公认的方法一样好，在 300 Hz 下评估时显示 66% 的灵敏度、100% 的特异性和 83% 的准确度。总之，建议通过阻抗谱基于 RhCE 电容变化来预测体内眼部刺激的定量分析。这种方法代表了一种非侵入性和非破坏性的替代方法，可以监测可逆效应或重复剂量毒性。建议通过阻抗谱基于 RhCE 电容的变化进行定量分析来预测体内眼部刺激。这种方法代表了一种非侵入性和非破坏性的替代方法，可以监测可逆效应或重复剂量毒性。建议通过阻抗谱基于 RhCE 电容的变化进行定量分析来预测体内眼部刺激。这种方法代表了一种非侵入性和非破坏性的替代方法，可以监测可逆效应或重复剂量毒性。