Graphene-based materials are not skin sensitizers: adoption of the in chemico / in vitro OECD test guidelines

The boost in the market size of graphene-based materials (GBMs) requires a careful evaluation of their impact on human health, acquiring robust and reliable data, also suitable for regulatory purposes. Considering cutaneous contact as one of the most relevant GBM exposure routes, this study is focused on skin sensitization, aimed at assessing the possibility to adopt the three in chemico/in vitro test guidelines (TGs) defined by the Organization for Economic Cooperation and Development (442C, D and E) to predict the first three phases of the skin sensitization adverse outcome pathway. Being originally validated for chemicals, modifications allowing their adoption for GBMs were evaluated. TG 442C was found to be not suitable for testing GBMs due to their reactivity, leading to possible misclassifications. In contrast, TG 442D and E can generally be applied for GBMs. However, protocol adjustments were required to assess cell viability reducing interferences for TG 442D, whereas caution should be exercised regarding dose-finding selection and GBM dispersion stability for TG 442E. When applying these modifications, GBMs were found to be unable to activate keratinocytes and promote dendritic cell differentiation, so they can be considered non-sensitizers. Overall, these results significantly contribute to understanding the safety profiles of GBMs and to improve testing methodologies to obtain reliable toxicological data.