Asbestos-exposed blood monocytes - deoxyribonucleic acid strand lesions in co-cultured bronchial epithelial cells

In Vitro Study of Mutagenesis Induced by Crocidolite-Exposed Alveolar Macrophages NR8383 in Cocultured Big Blue Rat2 Embryonic Fibroblasts

“…A system of separated coculture of NR8383 and Big Blue cells was used, similar to the system described by Kienast et al, which allows selective exposure of phagocytes to fibres [17]. NR8383 were exposed first to high concentrations of crocidolite or zymosan (up to 60  μ g/cm 2 ) for a short time (3 hours), conditions that normally favour the release of ROS from phagocytes.…”

“…When tested with another phagocyte type, the human primary blood polymorphonuclears, crocidolite was shown to produce a brief generation of free radicals at a concentration corresponding to 250 μ g/10 6 cells [ 41 , 42 ]. Human monocytes can similarly generate ROS after contact with amosite, chrysotile, ceramic, wool, or glass fibres [ 17 , 18 , 43 ]. In their coculture model, Kienast et al used primary human blood monocytes exposed to chrysotile (50 and 100 μ g/10 6 cells) as a “generator” system of reactive oxidative intermediates [ 17 ].…”

“…Human monocytes can similarly generate ROS after contact with amosite, chrysotile, ceramic, wool, or glass fibres [ 17 , 18 , 43 ]. In their coculture model, Kienast et al used primary human blood monocytes exposed to chrysotile (50 and 100 μ g/10 6 cells) as a “generator” system of reactive oxidative intermediates [ 17 ]. Consequently, both published data and our results indicate a poor capacity of rat alveolar macrophages to generate oxidative species when exposed in vitro to crocidolite.…”

“…The study of the gene expression of various inflammatory factors in NR8383 and Big Blue was a different approach used to assess the capacity of crocidolite to activate NR8383 and its effect on the cocultivated fibroblasts. A system of separated coculture of NR8383 and Big Blue cells was used, similar to the system described by Kienast et al, which allows selective exposure of phagocytes to fibres [ 17 ]. NR8383 were exposed first to high concentrations of crocidolite or zymosan (up to 60 μ g/cm 2 ) for a short time (3 hours), conditions that normally favour the release of ROS from phagocytes.…”

“…Coculture systems of phagocytes and epithelial cells have been described for mineral-fibre related toxicity or inflammation studies [ 16 – 19 ]. Using primary human blood monocytes cocultivated separately with bronchial epithelial cells, Kienast et al demonstrated that chrysotile phagocytosis resulted in the release of ROS in monocytes and induced DNA single-strand breaks in bronchial epithelial cells [ 17 ]. In a previous study, chrysotile phagocytosis was shown to induce the release of interleukin 1- β (IL1- β ), interleukin 6 (IL6), and tumor necrosis factor-alpha (TNF- α ) in both cocultivated cell types [ 20 ].…”