Three-Dimensional Cultured Human Nasal Epithelial Cell Model for Testing Respiratory Toxicity and Neurotoxicity of Air Pollutants

Accumulating evidence suggests a strong correlation between air pollution and neurological disorders; however, appropriate models and methodologies are currently lacking. In this study, a human nasal RPMI 2650 cell model based on air-liquid interface culture was discovered to possess olfactory epithelial cells. Two short-chain per- and polyfluoroalkyl substances (PFAS), PFBA and PFHxA, were used to validate the performance of the model. RNA sequencing initially revealed the adverse effects of two PFAS at environmentally relevant concentrations. Their effects on key nasal epithelial cell functions, including barrier protection, solute transport, and neuronal activity, were separately investigated. Both PFBA and PFHxA disrupted membrane integrity and increased cellular transport capacity, as indicated by the upregulation of ABC transporters. Additionally, they inhibited tight junction proteins, including ZO-1, claudin-3, and occludin, while increasing mucin expression and mucus secretion. PFHxA exhibited stronger effects in most assays and uniquely induced a significant upregulation of NOTCH1 expression (<i>p</i> < 0.05), highlighting its potential hazards on olfactory neurons. This study proposed a novel <i>in vitro</i> test model with the matched respiratory epithelial and neuronal end points, which was expected to improve toxicological research and risk assessment of air pollutants.