Understanding the impact of toluene exposure on lung epithelial cells is crucial for assessing industrial and environmental hazards. We investigated A549 cell responses to 10–100 ppm toluene in 2D (Transwell) and 3D (silica scaffold) cultures, comparing acute (direct exposure) and chronic (24-hour recovery) effects. GO and KEGG analyses revealed significant differences: 3D cultures exhibited enhanced cytoskeletal remodeling, mRNA processing, and metabolic adaptations, while 2D cultures showed stronger proliferative and mitotic activities. Chronic exposure resulted in sustained stress responses, particularly in DNA repair and oxidative stress pathways. Differential protein expression indicated that extracellular matrix interactions in 3D cultures drive distinct toxicity mechanisms. These findings highlight the importance of microenvironmental context in toxicological assessments and suggest 3D models as more physiologically relevant platforms. Validation through western blot, cell vaibility, and live-cell imaging supports our conclusions. This study provides critical insights into toluene-induced lung toxicity and offers a foundation for improved inhalation toxicity models.
ZHENG et al. (Wed,) studied this question.