The cytokinesis failure form cells with centrosome amplification, that could entry mitosis with multipolar spindles and complete or not cytokinesis, leading to aneuploid cells and/or multinucleation. All these studies suffer from the fact that they have no measured starting point of the asbestos fibre burden in the human lung tissue to compare their results to because of their cross-sectional design. Our results perfectly correlate to data published by Churg and dePaoli [21], who found that the ratio of chrysotile to amphibole (in this case tremolite) concentration did not change with time by comparing two sets of patients with a short and a long time interval to exposure cessation. The frequency of cells with two or more nuclei increased from about 15% in control cells to 45% in chrysotile-treated cells, reaching approximately 65% after longer recovery time (P < 0.01, Table 1). Am J Respir Crit Care Med. The image stacks were used for 3D reconstruction (B), evidencing the spatial interaction between cell and fiber, where the fiber seems to segment the nucleus. The long chrysotile fibers (> 20 μm) cleared quickly with T 1/2 estimated as 30 and 33 days, respectively in the brake dust and the chrysotile/brake dust exposure groups. PubMed  This data set was filtered for the following criteria (see also figure 1): 1) Results of asbestos body counts had to be available. In contrast, in control cells the frequency of multi-polar metaphases was around 5%. This study addresses the question whether the asbestos fibre burden in human lungs decreases with time after exposure cessation. 2004, 23: 3864-3873. The nuclear DNA content was quantified by image analysis with the software CIRES (Cell Image Retrieval and Evaluation System-Kontron Eletronik) installed in Axioskop microscope (Zeiss). However, the result of light microscopic counts are a good measure for the overall asbestos fibre burden of the lung tissue under investigation and may well serve for comparative analyses [30]. Nevins JR: E2F: a link between the Rb tumor suppressor protein and viral oncoproteins. However ninety to ninety five percent of all asbestos products manufactured have been made from chrysotile asbestos, from 1874 when asbestos was first used for industrial applications to the present. So the multinucleation observed after treatment could be a consequence of the direct fiber-chromatin interaction leading to its fragmentation, or a consequence of the cytokinesis failure resulting in cells with more than one nucleus. CAS  Together, data on the asbestos fibers potential to cause DNA damage [23] and the presence of fibers interacting with the chromatin in confocal images visualized by 3D reconstructions, suggest their association with centrosome amplification. Thus, exposure is not a confounder in the present data. HK2 cells treated with chrysotile for 48 h and recovered for additional 24 h in normal culture medium showed increased frequency of morphological nuclear alterations. By their pin-like/acicular structure the fibres are prone to get caught in the alveoli of the lung parenchyma, so that clearing them from the system cannot be achieved. Durnev AD, Daugel-Dauge NO, Korkina LG, Seredenin SB: Peculiarities of the clastogenic properties of chrysotile-asbestos fibers and zeolite particles. Nuclear DNA contents of mononucleated, binucleated and multinucleated control and treated cells were independently quantified by image analysis. The mutations might interfere in the mitotic checkpoints functions, but probably do not interfere in the cell-fiber interaction, like the fiber and cell cytoskeleton interaction and position of internalized fibers, as well as the process of fiber fragmentation. Conclusion. BAC maintained the cell culture, performed the chrysotile treatments, prepared the samples to microscope analyses, obtained the images and drafted the manuscript. BIA-Arbeitsmappe 26. Fukasawa K: Centrosome amplification, chromosome instability and cancer development. The present data show that 48 h chrysotile exposure can cause centrosome amplification, apoptosis and aneuploid cell formation even when long periods of recovery were provided. Trends Cell Biol. After that, the cells were washed three times with PBSA and incubated with 2nd antibody (anti-mouse FITC or Cy5, diluted 1:200) for 2 h. The nuclei were stained with propidium iodide and actin filaments with FICT-phalloidin for 20 min. Chrysotile 5R (Quebec Standard) obtained from SAMA Mineração de Amianto Ltda (Minaçu, GO, Brazil) were kindly provided by Dr. Flavia M. Cassiola. Intra-individual longitudinal data display: the asbestos fibre burden in living human lungs is stable over many years All authors read and approved the final manuscript. This article has supplementary material available from Science. Online ISSN: 1399-3003, Copyright © 2020 by the European Respiratory Society. In autopsy cases, analyses were performed in the upper and lower lobes of either side if possible. AB: asbestos bodies. Other evidence shows that chrysotile causes a much lower mesothelioma risk than amosite or crocidolite.12, 13 The rapid clearance of chrysotile from the lung with a half-life of a few months3 explains its virtual absence in our samples, and implies that we cannot estimate its effects except by noting that amphibole lung burdens account very well for mesothelioma incidence. 1989, 139: 885-90. The dettached and adherent cells were submitted to immunofluorescence independently. The centrosome amplification is classically associated with aneuploidy. 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