The biological interaction of several mineral fibers with cells and tissue can generate fibrosis, lung cancer, mesothelioma and represent an important interdisciplinary issue. These minerals are recognized as primarly factor in the generation of reactive oxygen species (ROS) [1]. The thought that only fiber morphology and dimension represent the carcinogenic factors sounds like naive at the today level of consciousness: it is evident that other aspects such as the availability of metallic cations, structural defects and surface characteristics have to be considered, as well. Aim of study was the mineralogical characterization of different asbestos fibres and the evaluation of their effects on biological systems. Methos. UICC chrysotile, chrysotile from Val Malenco, erionite from Nevada, UICC crocidolite fibers were characterized by Transmission Electron Microscopy with annexed Energy Dispersive Spectroscopy. To explore the possibility of different behaviour of mineral fibers, two biological environments (mesothelial and bronchoalveolar cells) representing their first target have been used. The fibres were administrated for 6, 12, 24 and 48h in Beas2B and MeT5A cells at 50μg/ml to evaluate their cytotoxic effects. Biofunctional parameters at time points were evaluated: % number of alive, death, apoptotic cells; % number of cells with low, medium, high ROS content. Analysis were conducted by Guava Easycite flow cytometer, 488 nm excitation wavelength (emissions recorded by the green channel for carboxy-DCF, the red and yellow channel for Via count dye; fluorescence intensity recorded on an average of 5,000 cells from each sample). Experiments were carried out at least in triplicates and results were analyzed by In-cyte software. DNA damage was evaluated by Comet assay. Results. The morphological and dimensional study assessed typical fibrous morphology for all the samples, their chemical composition match well with the expected for this minerals: both the chrysotile samples show the presence of aluminum and iron as substitute of tetrahedral and octahedral typical cations; the crocidolite bears an adding of calcium and the erionite has magnesium and iron cations normally unexpected in the general formula. All the fibers showed a high degree of crystallinity in the diffraction patterns study, without evidence of natural amorphization (e.g. weathering). Higher direct cytotoxic effects were exerted by UICC crocidolite and UICC chrysotile since short times of contact. Increases in DNA damage at 24h was evident in bronchial cells with erionite contact. Conclusions: the results of this work point out biological risks of erionite fibres and can be useful to the pathologists and biochemists who use asbestos minerals and fibrous erionite in-vitro studies as positive cyto- and geno-toxic standard references. [1] Schoonen M.A.A. et al. “Mineral-Induced Formation of Reactive Oxygen Species”, Reviews in Mineralogy & Geochemistry, Vol. 64, pp. 179-221, 2006, MS

BIOLOGICAL EFFECTS ON HUMAN CELLS OF CHARACTERIZED ASBESTOS FIBERS. AN IN VITRO STUDY / Pugnaloni, A.; Rippo, M. R.; Tiano, L.; Brugè, F.; Capella, S.; Vigliaturo, R.; Belluso., E.. - ELETTRONICO. - (2016), pp. 87-87. (Intervento presentato al convegno Medicina di Precisione Medicina Traslazionale Medicina di Laboratorio, tenutosi a Montesilvano (PE), Palacongressi d’Abruzzo nel 4-5-6 Ottobre 2016).

BIOLOGICAL EFFECTS ON HUMAN CELLS OF CHARACTERIZED ASBESTOS FIBERS. AN IN VITRO STUDY.

A. Pugnaloni
Membro del Collaboration Group
;
M. R. Rippo;L. Tiano;F. Brugè;
2016-01-01

Abstract

The biological interaction of several mineral fibers with cells and tissue can generate fibrosis, lung cancer, mesothelioma and represent an important interdisciplinary issue. These minerals are recognized as primarly factor in the generation of reactive oxygen species (ROS) [1]. The thought that only fiber morphology and dimension represent the carcinogenic factors sounds like naive at the today level of consciousness: it is evident that other aspects such as the availability of metallic cations, structural defects and surface characteristics have to be considered, as well. Aim of study was the mineralogical characterization of different asbestos fibres and the evaluation of their effects on biological systems. Methos. UICC chrysotile, chrysotile from Val Malenco, erionite from Nevada, UICC crocidolite fibers were characterized by Transmission Electron Microscopy with annexed Energy Dispersive Spectroscopy. To explore the possibility of different behaviour of mineral fibers, two biological environments (mesothelial and bronchoalveolar cells) representing their first target have been used. The fibres were administrated for 6, 12, 24 and 48h in Beas2B and MeT5A cells at 50μg/ml to evaluate their cytotoxic effects. Biofunctional parameters at time points were evaluated: % number of alive, death, apoptotic cells; % number of cells with low, medium, high ROS content. Analysis were conducted by Guava Easycite flow cytometer, 488 nm excitation wavelength (emissions recorded by the green channel for carboxy-DCF, the red and yellow channel for Via count dye; fluorescence intensity recorded on an average of 5,000 cells from each sample). Experiments were carried out at least in triplicates and results were analyzed by In-cyte software. DNA damage was evaluated by Comet assay. Results. The morphological and dimensional study assessed typical fibrous morphology for all the samples, their chemical composition match well with the expected for this minerals: both the chrysotile samples show the presence of aluminum and iron as substitute of tetrahedral and octahedral typical cations; the crocidolite bears an adding of calcium and the erionite has magnesium and iron cations normally unexpected in the general formula. All the fibers showed a high degree of crystallinity in the diffraction patterns study, without evidence of natural amorphization (e.g. weathering). Higher direct cytotoxic effects were exerted by UICC crocidolite and UICC chrysotile since short times of contact. Increases in DNA damage at 24h was evident in bronchial cells with erionite contact. Conclusions: the results of this work point out biological risks of erionite fibres and can be useful to the pathologists and biochemists who use asbestos minerals and fibrous erionite in-vitro studies as positive cyto- and geno-toxic standard references. [1] Schoonen M.A.A. et al. “Mineral-Induced Formation of Reactive Oxygen Species”, Reviews in Mineralogy & Geochemistry, Vol. 64, pp. 179-221, 2006, MS
2016
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11566/266428
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