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Effects of carbon black nanoparticles on human pulmonary cell lines and precision cut lung slices

: Hansen, Tanja; Kopf, Johannes Claudius; Danov, Olga; Ströbele, M.; Braun, Armin; Sewald, Katherina; Steinberg, P.; Fehrenbach, H.

The Toxicologist 52 (2013), Nr.1, S.95, Abstract PS 444
ISSN: 0731-9193
Society of Toxicology (Annual Meeting) <52, 2013, San Antonio/Tex.>
Zeitschriftenaufsatz, Konferenzbeitrag
Fraunhofer ITEM ()

Carbon black nanoparticles (CBNPs) are among the most abundantly used nanomaterials and have been reported to cause adverse health effects after inhalation exposure. The aim of this study was to compare the effects of Printex® 90 and acetylene soot particles in human pulmonary cell lines (16HBE14o-, Calu-3, A549) and precision cut lung slices (PCLS) of mice, rats and humans using a wide concentration range. Particle size distribution in the cell culture medium was determined by dynamic light scattering. Viability assays were LIVE/DEAD® staining and WST-1 assay for PCLS and WST-8 and neutral red assay for cell lines. CBNP-induced formation of reactive oxygen species (ROS) was assessed in A549 and 16HBE14ocells by flow cytometry using the DCFH-DA assay. Furthermore, the effect of CBNP exposure on the transepithelial electrical resistance (TEER) was investigated in Calu-3 cells after 24, 48 and 120h treatment with 10 and 50 g/ml CBNPs.
With PCLS, the inflammatory response was assessed by measuring pro-inflammatory cytokines (i.e. IL-1alpha, TNF- alpha, IL-8). Both CBNPs tested were not toxic in physiologically relevant concentrations. Significant cytotoxicity was observed in the WST-8 assay for both CBNPs at 50 g/ml after 48h, whereas no effects were found in the neutral red assay. Increased ROS formation was observed with both CBNPs after 24 and 48 h. Interestingly, acetylene soot particles cause significant TEER reduction at both dose levels and all time points tested whereas Printex® 90 reduced the TEER only after 120h at the high dose. Neither Printex® 90 nor acetylene soot particles induced the secretion of proinflammatory cytokines in mouse and rat PCLS. In conclusion, the combination of in vitro and ex vivo models provides a valuable tool to assess the acute irritation and inflammation effects of CBNPs on lung tissue.