Because its critical location, the skin is the main target of environmental stressors such as ozone (O3). Although O3 does not penetrate the deeper layers of skin it is able to react readily with stratum corneum lipids [1]. The toxic effects of O3 on the uppermost layers, induced either directly by the oxidation of biomolecules or by driving the radical-dependent production of cytotoxic, non-radical species (aldehydes), have repercussions on deeper cellular layers, triggering a cascade of cellular stress and inflammatory responses that can lead to skin pathologies [2] and [3]. Furthermore, O3 is able to induce the depletion of cutaneous antioxidants [4]. Therefore, topical application of antioxidants could prevent pollution induces cutaneous damages. In our recent work, the use of topical antioxidant mixtures (MIXs) has proven an effective defensive approach against O3-induced oxidative damage in human keratinocytes [5]. In the present study, which is a methodological extension of the one recently published [5], we evaluated the ability of the three MIXs to prevent the noxious effects of O3 on a customized reconstructed human epidermis (RHE, Episkin), where sebum was applied to reproduce a model resembling “in vivo” expectations [6