An In Vitro Lung System to Assess the Proinflammatory Hazard of Carbon Nanotube Aerosols

July 27, 2020


Hana Barosova 1,2 , Bedia Begum Karakocak 1 , Dedy Septiadi 1 , Alke Petri-Fink 1,3, Vicki Stone 4 and Barbara Rothen-Rutishauser 1,
1 BioNanomaterials Group, Adolphe Merkle Institute, University of Fribourg, 1700 Fribourg, 
2 Institute of Experimental Medicine of the Czech Academy of Sciences, 142 20 Prague, Czech Republic
3 Department of Chemistry, University of Fribourg, 1700 Fribourg, Switzerland
4 Institute of Biological Chemistry, Biophysics and Bioengineering, Heriot-Watt University, Edinburgh EH14 4AS, UK

The in vitro co-culture model consisting of three human cell lines were exposed at the ALI using the VITROCELL® Cloud system, equiped with QCM allowing to measure and record the deposited dose online. The nebulizer was ideal for the aerosols to suffciently mix within the entire chamber, hence resulting in uniform droplet deposition. This study shows, that this model is not limited to testing potentially hazardous nanomaterials to human cell line co-culture models.



In vitro three-dimensional (3D) lung cell models have been thoroughly investigated in recent years and provide a reliable tool to assess the hazard associated with nanomaterials (NMs) released into the air. In this study, a 3D lung co-culture model was optimized to assess the hazard potential of multiwalled carbon nanotubes (MWCNTs), which is known to provoke inflammation and fibrosis, critical adverse outcomes linked to acute and prolonged NM exposure. The lung co-cultures were exposed to MWCNTs at the air-liquid interface (ALI) using the VITROCELL® Cloud system while considering realistic occupational exposure doses. The co-culture model was composed of three human cell lines: alveolar epithelial cells (A549), fibroblasts (MRC-5), and macrophages (di erentiated THP-1). The model was exposed to two types of MWCNTs (Mitsui-7 and Nanocyl) at di erent concentrations (2–10 µg/cm2) to assess the proinflammatory as well as the profibrotic responses after acute (24 h, one exposure) and prolonged (96 h, repeated exposures) exposure cycles. The results showed that acute or prolonged exposure to di erent concentrations of the tested MWCNTs did not induce cytotoxicity or apparent profibrotic response; however, suggested the onset of proinflammatory response.

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