Pre-validation of a novel reconstructed skin equivalent model for skin irritation and nanoparticle risk assessment

Abstract

In alignment with the global movement toward reducing animal testing, several reconstructed human epidermis (RHE) models have been created for conducting skin irritation tests. These models have undergone development, verification, validation, and integration into OECD TG 439. Our team has introduced a novel in-house RHE named GB-RHE, and we adhere to OECD TG 439 to pre-validate the model and test its potential employment for nanoparticle irritation studies. GB-RHE exhibits morphological, biochemical, and physiological attributes equivalent to the human epidermis, featuring well-differentiated multilayered viable keratinocytes with a robust barrier function. The performance of the GB-RHE model was evaluated using ten reference chemicals, following the performance standard of OECD TG 439. The results demonstrated commendable predictive capacity and showed that titanium dioxide nanoparticles (TiO₂ NPs) are ‘non-irritant’ to the human epidermis following the globally harmonized classification system. However, although the histological analysis did not show morphological changes, transmission electron micrographs demonstrated that TiO₂ NPs can be internalized, reaching the external viable layers of the epidermis. This study demonstrates that in addition to the potential of the GB-RHE model to evaluate skin irritation, this model also has the potential to evaluate the skin toxicity of NPs and carry out cell internalization studies.