https://doi.org/10.1016/j.tiv.2024.105841
Vegard Sæter Grytting a, Tonje Skuland a, Jarle Ballangby a, Magne Refsnes a, Marit Låg a, Johan Øvrevik a b, Espen Mariussen a
aDepartment of Air quality and Noise, Division of Climate and Environmental Health, Norwegian Institute of Public Health, PO Box 222, Skøyen, Oslo 0213, Norway
bDepartment of Biosciences, Faculty of Mathematics and Natural Sciences, University of Oslo, PO Box 1066 Blindern, 0316 Oslo, Norway
Highlights
•3D cell cultures based on Calu-3 cells demonstrated superior barrier capacity to those based on A549 and HBEC3-KT cells.
•Exposure to fine PM at ALI induced pro-inflammatory responses in all cell models.
•The cytokine responses depended on the addition of macrophages to the apical compartment.
•Cellular responses in the basolateral compartment varied depending on the barrier capacity of the models.
Abstract
3D cell culture models exposed at the air-liquid interface (ALI) represent a potential alternative to animal experiments for hazard and risk assessment of inhaled compounds. This study compares cocultures composed of either Calu-3, A549 or HBEC3-KT lung epithelial cells, cultured together with THP-1-derived macrophages and EA.hy926 endothelial cells, in terms of barrier capacity and responses to a standard reference sample of fine particulate matter (SRM 2786). High-content imaging analysis revealed a similar cellular composition between the different cell models. The 3D cell cultures with Calu-3 cells showed the greatest barrier capacity, as measured by transepithelial electrical resistance and permeability to Na-fluorescein. Mucus production was detected in 3D cell cultures based on Calu-3 and A549 cells. Exposure to SRM 2786 at ALI increased cytokine release and expression of genes associated with inflammation and xenobiotic metabolism. Moreover, the presence of THP-1-derived macrophages was central to the cytokine responses in all cell models. While the different 3D cell culture models produced qualitatively similar responses, more pronounced pro-inflammatory responses were observed in the basolateral compartment of the A549 and HBEC3-KT models compared to the Calu-3 model, likely due to their reduced barrier capacity and lower retention of secreted mediators in the apical compartment.
