https://link.springer.com/article/10.1186/s12302-025-01258-8
A. Zimmermann-Klemd, L. Wende Wolf, FS. Emser, J. Daniel, M. Follo, R. Trittler, B. Rothen-Rutishauser, P. Deibert, C. Gründemann & M. Garcia-Käufer
Institute for Exercise- and Occupational Medicine, Medical Center – University of Freiburg, Faculty of Medicine, University of Freiburg, Germany
The study aimed to verify the performance of the high-throughput VITROCELL® 96 Exposure System, a high‑throughput continuous‑flow air–liquid interface (ALI) platform for comparative hazard assessment of complex aerosols.
A549 alveolar epithelial cells cultured on 96‑well HTS inserts at the ALI were exposed to whole aerosols from 1R6F reference cigarette smoke and IQOS heated tobacco products. Aerosols were generated using the VITROCELL® VC 1 Smoking Machine and delivered to the exposure module via a 12‑column dilution concept, providing eleven aerosol concentrations plus clean air controls in a single experiment. Dosimetry based on total particulate matter and glycerol confirmed homogeneous aerosol distribution and close agreement between nominal and delivered doses across all 96 exposure positions. A tiered assay battery integrating cytotoxicity, genotoxicity (comet assay and in situ γ‑H2AX) and immunomodulatory readouts demonstrated robust, concentration‑dependent responses with minimal vehicle‑related background and high intra‑ and inter‑experiment reproducibility. Benchmark‑dose modelling enabled derivation of points of departure and relative potency factors, revealing markedly lower cytotoxic, genotoxic and immunomodulatory potency for IQOS aerosol compared with 1R6F smoke, while confirming biological activity of both emissions. Collectively, the work establishes the VITROCELL® 96 Exposure System as a next‑generation, high‑throughput solution for early hazard ranking and mechanistic profiling of inhaled aerosols, supporting its use as a core technology within new approach methodologies (NAMs) in inhalation toxicology.
