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In certain situations, such as aging, the ETCs functionality may decline, leading to an excessive diversion of electrons from the ETC pathway, which then react with O 2 to generate significant amounts of O 2 -, the precursors to most ROS (Nilsson and Tarnopolsky, 2019

The current standard operating procedure by the European Center for the Validation of Alternative Methods (ECVAM) recommends the use of MTT reduction as parameter to make prediction on the skin irritancy of substances using EpiSkin TM test method, while IL-1 endpoint is regarded as a useful adjunct to MTT assay 48