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The purpose of this work is to define a sample preparation protocol that allows inorganic fibers and particulate matter extracted from different biological samples to be characterized morphologically, crystallographically and chemically by transmission electron microscopy-energy dispersive spectroscopy (TEM-EDS). The method does not damage or create artifacts through chemical attacks of the target material. A fairly rapid specimen preparation is applied with the aim of performing as few steps as possible to transfer the withdrawn inorganic matter onto the TEM grid. The biological sample is previously digested chemically by NaClO. The salt is then removed through a series of centrifugation and rinse cycles in deionized water, thus drastically reducing the digestive power of the NaClO and concentrating the fibers for TEM analysis. The concept of equivalent hydrodynamic diameter is introduced to calculate the settling velocity during the centrifugation cycles. This technique is applicable to lung tissues and can be extended to a wide range of organic materials. The procedure does not appear to cause morphological damage to the fibers or modify their chemistry or degree of crystallinity. The extrapolated data can be used in interdisciplinary studies to understand the pathological effects caused by inorganic materials.
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Amiantos Anfibólicos/análise , Asbestos Serpentinas/análise , Manejo de Espécimes/métodos , Amiantos Anfibólicos/química , Asbestos Serpentinas/química , Humanos , Pulmão , Microscopia Eletrônica de Transmissão , Hipoclorito de Sódio/químicaRESUMO
Because asbestos diseases represent a complex pattern of legal, social, and political issue, the involvement of the mineralogist and pathologist for a multidisciplinary assessment of its diagnosis helps investigate the relationship between mesothelioma or lung cancer and occupational or environmental asbestos exposure.In the present study, we consider the concentrations of asbestos bodies (ABs) detected by optical microscopy (OM) and scanning electron microscopy (SEM) and the burden of different kinds of mineral fibers (among which is asbestos) identified by SEM combined with an energy dispersive spectrometer (EDS), in 10 lung tissue samples of subjects with occupational and nonoccupational exposure to asbestos.In all subjects with occupational exposure to asbestos, more than 1000 ABs per gram of dry weight were detected both with OM and SEM; this concentration is internationally accepted as suggesting high probability of past occupational exposure to asbestos.In 9 lung samples of the 10 investigated by SEM-EDS different inorganic fibers were found. Asbestos fibers have been identified too, and more than 100,000 fibers per gram of dry weight were detected in subjects with occupational exposure; this concentration is internationally accepted as suggesting high probability of past occupational exposure to asbestos.Instead, when the ABs burden is low or moderate (such as in subjects with absent or probable asbestos exposure), the correlation between ABs concentration determined by OM and those determined by SEM is lost. Therefore, when the ABs value in OM is borderline, the SEM investigation became essential. Furthermore, the mineralogical analysis by SEM-EDS (identification and quantification of inorganic fibers in general and asbestos in particular) of the fibers detected in the lung tissues is very useful, if not necessary, to complete the pathological diagnosis of asbestos-related malignancies in medicolegal field.
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Amianto/análise , Asbestose/diagnóstico , Neoplasias Pulmonares/diagnóstico , Mesotelioma/diagnóstico , Neoplasias Pleurais/diagnóstico , Humanos , Pulmão/química , Pulmão/patologia , Pulmão/ultraestrutura , Microscopia Eletrônica de Varredura , Fibras Minerais/análise , Estudos Retrospectivos , Espectrometria por Raios XRESUMO
BACKGROUND: Italian law requires an extensive health surveillance of workers after cessation of their employment status in the case of occupational exposure to carcinogens, including asbestos. Nonetheless, Italian law does not specify the timeframe of these clinical checks, nor who has financial and organizational responsibility for this surveillance. A literature search confirmed a lack of consensus around the objectives and methods to follow up workers with past occupational exposure to asbestos. OBJECTIVES: To develop an updated evidence-based methodology for an appropriate health surveillance programme. METHODS: We present an overview of the field experience developed by the Veneto Region from 2000 to 2011, and new studies that could contribute to establishing a national policy for the medical surveillance of workers with past asbestos exposure. RESULTS: There were three specific topics: (1) definition of a reliable method to identify asbestos workers (through multiple sources and procedures that meet current confidentiality regulations); (2) detection of asbestos fibres in biological media (to support the etiological diagnosis of asbestos-related diseases); (3) creation of a national protocol of health surveillance (through the assessment of policies developed by other Regions in this field, and recruiting from these regions a cohort of past-exposed workers: the epidemiological study should offer relevant suggestions for specific surveillance approaches, based on either estimated cumulative asbestos exposure or detection of x-ray patterns of pleural plaques and/or asbestosis). CONCLUSIONS: These studies will support the Regions in setting up health care policies directed at workers with past asbestos exposure.
Assuntos
Amianto/efeitos adversos , Asbestose/epidemiologia , Exposição Ocupacional , Vigilância da População , Amianto/análise , Asbestose/sangue , Asbestose/etiologia , Biomarcadores , Detecção Precoce de Câncer/economia , Detecção Precoce de Câncer/métodos , Seguimentos , Política de Saúde , Humanos , Itália , Responsabilidade Legal , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/economia , Neoplasias Pulmonares/epidemiologia , Neoplasias Pulmonares/etiologia , Neoplasias Pulmonares/prevenção & controle , Masculino , Mesotelioma/diagnóstico , Mesotelioma/economia , Mesotelioma/epidemiologia , Mesotelioma/etiologia , Mesotelioma/prevenção & controle , Pessoa de Meia-Idade , Fibras Minerais/análise , Ocupações , Osteopontina/sangue , Neoplasias Pleurais/diagnóstico , Neoplasias Pleurais/economia , Neoplasias Pleurais/epidemiologia , Neoplasias Pleurais/etiologia , Neoplasias Pleurais/prevenção & controle , Vigilância da População/métodos , Avaliação de Programas e Projetos de Saúde , Reprodutibilidade dos Testes , Testes de Função Respiratória , Aposentadoria , Estudos Retrospectivos , FumarRESUMO
Asbestos occurrence has been mainly monitored in air so far and only limitedly considered in other matrices, such as water. Waterborne asbestos could originate from natural or anthropogenic sources, leading to non-conventional exposure scenarios. It could be a secondary source of airborne asbestos in case of water-to-air migration, particularly in case of surface moving water, such as in rivers and streams. The scarce attention dedicated to waterborne asbestos has led to a considerable fragmentation in regulatory approaches regarding the study of water samples possibly contaminated by mineral fibres. In this context, this study has been designed to test the reliability of an existing analytical method devoted to natural waters investigations. Following the operational protocol issued by the Piedmont (Italy) Environmental Protection Agency, Scanning Electron Microscopy analyses have been performed on a standard sample of waterborne chrysotile, mimicking stream water. The investigations have been performed by different operators and using different analytical setups, to verify whether the method applied has a good interlaboratory reproducibility and which could be the most error-prone analytical steps. Three data sets have been obtained on the same sample, showing a low reproducibility among each other. Possible reasons causing this discrepancy have been discussed in detail and good practices to perform reliable analyses on surface water samples containing asbestos have been proposed to help the regulatory organs to better define analytical protocols.
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Asbestos is widely recognized as being a carcinogen when dispersed in air, but very little is known about its exposure pathways in water and its subsequent effects on human health. Several studies have proved asbestos presence in groundwater but failed to assess its mobility in aquifer systems. This paper aims to fill this gap by studying the transport of crocidolite, an amphibole asbestos, through sandy porous media mimicking different aquifer systems. To this purpose, two sets of column test were performed varying the crocidolite suspension concentration, the quartz sand grain size distribution, and the physicochemical water parameters (i.e., pH). The results proved that crocidolite is mobile in quartz sand due to the repulsive interactions between fibres and porous media. The concentration of fibres at the outlet of the column were found to decrease when decreasing the grain size distribution of the porous medium, with a bigger impact on highly concentrated suspensions. In particular, 5-to-10-µm-long fibres were able to flow through all the tested sands while fibres longer than 10 µm were mobile only through the coarser medium. These results confirm that groundwater migration should be considered a potential exposure pathway while implementing human health risk assessment.
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Background: In Italy the incidence of malignant mesothelioma (MM) among women is remarkably high, due to the several contexts in which women had been exposed to asbestos. However, very few studies in literature focus on the inorganic lung content in women. The aim of this retrospective, observational study is to investigate the asbestos lung burden, in terms of concentration, dimensions and type of asbestos, in 42 women who died from MM and had been non-occupationally exposed to asbestos during the activity of the asbestos-cement plant located in Broni (Pavia, Northern Italy) where mainly chrysotile, crocidolite and amosite were used. Methods: Lung samples taken during forensic autopsies have been digested using sodium hypochlorite and filtered through a cellulose-ester membrane. The filter was examined using a scanning electron microscope and the chemical composition of the fibers was analyzed using an electron dispersive spectroscopy. The number of detected inorganic fibers, asbestos fibers and asbestos bodies (ABs) were normalized to 1 gram of dry tissue. Results: In six samples no asbestos has been detected. Overall, the most represented kind of asbestos was amosite, followed by crocidolite, tremolite/actinolite asbestos and chrysotile. The concentration of all inorganic fibers was significantly higher in women with environmental and household exposures compared with those with only environmental exposure (P=0.025), as well as the concentration of asbestos fibers (P=0.019) and ABs (P=0.049). We found a significant correlation between the concentration of asbestos fibers and the duration of exposure (rho =0.413, P=0.008), as well as with the latency of MM (rho =0.427, P=0.005). The distance of the residential address from the factory and the time spent daily in contact with asbestos did not influence the lung asbestos burden. Conclusions: These results suggest the relevance of the lung clearance of asbestos, regarding mainly chrysotile. As a consequence, although scanning electron microscopy -energy dispersive X-ray spectroscopy (SEM-EDS) is considered the most reliable tool for assessing previous exposure to asbestos, its results should be interpreted with caution, especially in a legal context. In addition, our data confirm the relevance of environmental and household exposure in determining asbestos concentration in lungs and highlight the importance of household exposure.
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The environmental impact of natural occurrences of asbestos (NOA) and asbestos-like minerals is a growing concern for environmental protection agencies. The lack of shared sampling and analytical procedures hinders effectively addressing this issue. To investigate the hazard posed by NOA, a multidisciplinary approach that encompasses geology, mineralogy, chemistry, and toxicology is proposed and demonstrated here, on a natural occurrence of antigorite from a site in Varenna Valley, Italy. Antigorite is, together with chrysotile asbestos, one of the serpentine polymorphs and its toxicological profile is still under debate. We described field and petrographic analyses required to sample a vein and to evaluate the NOA-hazard. A combination of standardized mechanical stress and automated morphometrical analyses on milled samples allowed to quantify the asbestos-like morphology. The low congruent solubility in acidic simulated body fluid, together with the toxicity-relevant surface reactivity due to iron speciation, signalled a bio-activity similar or even greater to that of chrysotile. Structural information on the genetic mechanism of antigorite asbestos-like fibres in nature were provided. Overall, the NOA site was reported to contain veins of asbestos-like antigorite and should be regarded as source of potentially toxic fibres during hazard assessment procedure.
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There are six elongate mineral particles (EMPs) corresponding to specific dimensional and morphological criteria, known as asbestos. Responsible for health issues including asbestosis, and malignant mesothelioma, asbestos has been well researched. Despite this, significant exposure continues to occur throughout the world, potentially affecting 125 million people in the workplace and causing thousands of deaths annually from exposure in homes. However, there are other EMPS, such as fibrous/asbestiform erionite, that are classified as carcinogens and have been linked to cancers in areas where it has been incorporated into local building materials or released into the environment through earthmoving activities. Erionite is a more potent carcinogen than asbestos but as it is seldom used for commercial purposes, exposure pathways have been less well studied. Despite the apparent similarities between asbestos and fibrous erionite, their health risks and exposure pathways are quite different. This article examines the hazards presented by EMPs with a particular focus on fibrous erionite. It includes a discussion of the global locations of erionite and similar hazardous minerals, a comparison of the multiple exposure pathways for asbestos and fibrous erionite, a brief discussion of the confusing nomenclature associated with EMPs, and considerations of increasing global mesothelioma cases.
Assuntos
Amianto , Asbestose , Mesotelioma Maligno , Mesotelioma , Zeolitas , Amianto/toxicidade , Asbestose/epidemiologia , Carcinógenos/toxicidade , Humanos , Mesotelioma/induzido quimicamente , Mesotelioma/epidemiologiaRESUMO
In Naturally Occurring Asbestos (NOA) rich areas, water flows through asbestos bearing rocks and soils and generates waterborne fibres that may migrate in air and become a risk for humans. Research on the migration and dispersion after water vaporisation has been so far only marginally evaluated. This study investigates the migration in air of asbestos from a set of suspensions contaminated by chrysotile from Balangero (Italy), under controlled laboratory conditions. We evaluated i) the morphological modifications that might occur to chrysotile during migration from water to air, and ii) the amount of airborne chrysotile mobilised from standardised suspensions. Morphological alteration of asbestos fibres occurred during water-air migration and impacted on the analytical response of electron microscopy. Waterborne asbestos concentration higher than 40 â 106 f/L generates in air concentration higher than 1 fibre per litre [f/L], the alarm threshold limit set by World Health Organization for airborne asbestos. A possible correlation between the waterborne fibre concentration as mass or number of fibres per volume unit [µg/L or f/L] was observed.
Assuntos
Amianto , Exposição Ocupacional , Asbestos Serpentinas , Humanos , Itália , Exposição Ocupacional/análise , ÁguaRESUMO
The number of the Asbestos Bodies (AB), i.e. asbestos that developed an iron-protein coating during its permanence in biological tissues, is one of the most accessible markers of asbestos exposure in individuals. The approaches developed to perform AB count in biological tissues are based on the manual examination of tissue digests or histological sections by means of light or electron microscopies. Although these approaches are well established and relatively accessible, manual examination is time-consuming and can be reader-dependent. Besides, approximations are applied because of the limitations of 2D readings and to speed up manual counts. In addition, sample preparation using tissue digests require an amount of tissue that can only be obtained by invasive surgery or post-mortem sampling. In this paper, we propose a new approach to AB counting based on non-destructive 3D imaging, which has the potential to overcome most of the limitations of conventional approaches. This method allows automating the AB count and determining their morphometry distribution in bulk tissue samples (ideally non-invasive needle biopsies), with minimal sample preparation and avoiding approximations. Although the results are promising, additional testing on a larger number of AB-containing biological samples would be required to fully validate the method.
Assuntos
Amianto/efeitos adversos , Pulmão/diagnóstico por imagem , Pulmão/patologia , Idoso de 80 Anos ou mais , Feminino , Humanos , Imageamento Tridimensional , Masculino , Microtomografia por Raio-XRESUMO
We appreciate very much the interest of Mirabelli et al. [...].
Assuntos
Amianto , Mesotelioma , Exposição Ocupacional , Amianto/toxicidade , Humanos , Itália/epidemiologia , Pulmão , Mesotelioma/induzido quimicamente , Mesotelioma/epidemiologia , Exposição Ocupacional/efeitos adversos , Saúde PúblicaRESUMO
In the lungs, asbestos develops an Fe-rich coating (Asbestos Body, AB) that becomes the actual interface between the foreign fibers and the host organism. Conventional approaches to study ABs require an invasive sample preparation that can alter them. In this work, a novel combination of x-ray tomography and spectroscopy allowed studying unaltered lung tissue samples with chrysotile and crocidolite asbestos. The thickness and mass density maps of the ABs obtained by x-ray tomography were used to derive a truly quantitative elemental analysis from scanning x-ray fluorescence spectroscopy data. The average mass density of the ABs is compatible with that of highly loaded ferritin, or hemosiderin. The composition of all ABs analyzed was similar, with only minor differences in the relative elemental fractions. Silicon concentration decreased in the core-to-rim direction, indicating a possible partial dissolution of the inner fiber. The Fe content in the ABs was higher than that possibly contained in chrysotile and crocidolite. This finding opens two opposite scenarios, the first with Fe coming from the fiber bulk and concentrating on the surface as long as the fiber dissolves, the second where the Fe that takes part to the formation of the AB originates from the host organism Fe-pool.
Assuntos
Amianto/química , Asbestose/diagnóstico por imagem , Pulmão/diagnóstico por imagem , Espectrometria de Fluorescência/métodos , Tomografia por Raios X/métodos , Idoso de 80 Anos ou mais , Feminino , Humanos , MasculinoRESUMO
Biodurability is one of the main determinants of asbestos hazardousness for human health. Very little is known about the actual persistence of asbestos in lungs and its clearance, nor about differences in this regard between the different mineralogical types of asbestos. The aim of the present study was to evaluate the amount, the dimensional characteristics and the mineralogic kinds of asbestos in lungs (measured using SEM-EDS) of a series of 72 deceased subjects who were certainly exposed to asbestos (mainly crocidolite and chrysotile) during their life. Moreover, we investigated possible correlations between the lung burden of asbestos (in general and considering each asbestos type), as well as their dimension (length, width, and l/w ratio) and the duration of exposure, the latency- in case of malignant mesothelioma (MM), the survival and the time since the end of exposure. In 62.5% of subjects, asbestos burden in lungs was lower that the threshold considered demonstrative for occupational exposure. In 29.1% of cases no asbestos was found. Chrysotile was practically not detected. The mean length of asbestos fibers and the length to width ratio were significantly related to the duration of exposure to asbestos. No other statistically significant correlations were found between the amount and dimensional characteristics of asbestos (nor with the relative amount of each asbestos type) and the other chronological variables considered. In conclusion, it was pointed out that chrysotile can be completely removed from human lungs in <8 years and, instead, amphiboles persist much more time. The present results suggest, as well, that the finding of no asbestos in lungs cannot rule out the attribution of MM to asbestos (in particular, chrysotile) inhaled in an occupational setting. This point is of crucial importance from a legal point of view.
Assuntos
Amianto , Neoplasias Pulmonares , Amianto/efeitos adversos , Amiantos Anfibólicos/efeitos adversos , Asbestos Serpentinas/efeitos adversos , Humanos , Pulmão , Neoplasias Pulmonares/induzido quimicamenteRESUMO
Increased mortality due to malignant mesothelioma has been demonstrated by several epidemiologic studies in the area around Broni (a small town in Lombardy, northern Italy), where a factory producing asbestos cement was active between 1932 and 1993. Until now, the inorganic fiber burden in lungs has not been investigated in this population. The aim of this study is to assess the lung fiber burden in 72 individuals with previous occupational and/or anthropogenic environmental exposure to asbestos during the activity of an important asbestos cement factory. Inorganic fiber lung burden was assessed in autoptic samples taken from individuals deceased from asbestos-related diseases using a scanning electron microscope equipped with an energy-dispersive spectrometer. Significant differences in the detected amount of asbestos were pointed out among the three types of exposure. In most lung samples taken from patients who died of mesothelioma, very little asbestos (or, in some cases, no fibers) was found. Such subjects showed a significantly lower median amount of asbestos as compared to asbestosis. Almost no chrysotile was detected in the examined samples. Overall, crocidolite was the most represented asbestos, followed by amosite, tremolite/actinolite asbestos, and anthophyllite asbestos. There were significant differences in the amount of crocidolite and amosite fibers according to the kind of exposure. Overall, these findings provide novel insights into the link between asbestos exposure and mesothelioma, as well as the different impacts of the various types of asbestos on human health in relation to their different biopersistences in the lung microenvironment.
Assuntos
Amianto , Neoplasias Pulmonares , Mesotelioma , Exposição Ocupacional , Exposição Ambiental , Humanos , Itália/epidemiologia , Pulmão , Neoplasias Pulmonares/epidemiologia , Neoplasias Pulmonares/etiologia , Mesotelioma/induzido quimicamente , Mesotelioma/epidemiologia , Exposição Ocupacional/efeitos adversos , Microambiente TumoralRESUMO
The present investigation represents a new approach useful to evaluate the general population risk correlated with environmental exposure to air dispersed inorganic fibers. The used method is based on the evaluation of the respirable inorganic fibers both air dispersed in a big city and contained in lungs of the general population following their respiration. Moreover, these data allow to identify the sources of dispersion (anthropogenic or natural) in air of the inorganic fibers and therefore to apply strategies to improve air quality. To describe this approach, we investigated air samples from a big city in NW Italy and lung inorganic burden of people here lived. This paper reports the data of the airborne inorganic fibers detected in two sampling campaign (2014 and 2016), in 24 districts of Torino (Piemonte - NW Italy), and in some autoptic lungs of general population lived here. The airborne fibers (collected on mixed-cellulose esters membrane) were characterized by SEMEDS. The identified inorganic fiber species were assigned to 5 classes, one of these including 2 types of asbestos. These last are grouped as tremolite/actinolite asbestos. They are dispersed from natural sources (i.e. certain kinds of rocks outcropping in the city surrounding areas). In no-one of the 24 districts of Torino their concentration highlighted a situation of asbestos pollution in place. A correlation with inorganic fibers (collected on mixed-cellulose esters membrane and characterized by SEM-EDS) detected in lung tissue samples of 10 subjects lived in Torino all their life and without professional exposure to asbestos were attempted. The only types of fibers identified as asbestos are tremolite/actinolite asbestos, and they match those detected in air sampling. The number of fibers per 1 g of tissue dry weight is lower than the quantities reported as indicative of significant asbestos exposure. We observed interesting gender differences.
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Poluição do Ar , Amianto , Cidades , Humanos , Itália , Pulmão/químicaRESUMO
Professional exposure to asbestos fibres is widely recognized as very dangerous to human health and for this reason many countries have banned their commercial uses. People, nevertheless, continue to be exposed to low dose of asbestos from natural and anthropogenic sources still in loco, for which the potential hazard is unknown. The aim of this research is to assess environmental exposure in an area with outcropping serpentinite rocks, which bear asbestos mineralizations, using sentinel animals which are a non-experimental animal model. We studied the burden of inorganic fibres in cattle lungs which come from two areas in Italy's Western Alps bearing serpentinitic outcrops: Susa Valley with a heavy anthropization and Lanzo Valleys, with a minor human impact. The identification and quantification of inorganic fibres were performed by Scanning Electron Microscope (SEM) and Energy Dispersive Spectrometer (EDS). In comparison to humans, studies of animals have some advantages, such as no occupational exposure or history of smoking and, in the case of cattle, a sedentary life restricted to one region. Results spotlight that over than 35% of inorganic fibres found both in Susa and Lanzo valleys, belong to asbestos mineralogical species (asbestos tremolite/actinolite, chrysotile s.s., asbestos grunerite, crocidolite). We also observed a higher concentration of artificial fibrous products in Susa samples showing a correlation with the level of anthropization. These results confirm that sentinel animals are an excellent model to assess breathable environmental background because it is possible to eliminate some variables, such as unknown occupational exposure.
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Amianto/toxicidade , Exposição Ambiental , Compostos Inorgânicos/toxicidade , Pulmão/patologia , Altitude , Animais , Asbestos Serpentinas/toxicidade , Bovinos , Feminino , Geografia , Humanos , Itália , Pulmão/ultraestrutura , Masculino , Microscopia Eletrônica de Varredura , Modelos AnimaisRESUMO
Mesothelioma is a tumor of the serosal membranes described both in human and veterinary medicine. While in humans the relationship between mesothelioma and exposure to asbestos and some other asbestiform minerals is well known, in animals it is still difficult to establish. In this paper a case of malignant peritoneal mesothelioma probably related to asbestos exposure in a wild boar is described. At post-mortem evaluation the peritoneum, diaphragm and serosal surface of liver and kidneys showed isolated to coalescent multiple nodular lesions. Samples from diaphragm, liver and lung were collected to perform microbiological and histological investigations. To assess the presence of asbestos and/or other asbestiform minerals, SEM-EDS investigations were performed on organs and soil samples collected from the area where the wild boar lived. Microbiological investigations were negative for Mycobacterium species. Gross and histological examination were compatible with a biphasic mesothelioma, with nodules composed of epithelioid and sarcomatoid elements with high pleomorphism. Immunohistochemistry revealed only multifocal scattered positivity for WT-1 and D2-40. Asbestos fibres were detected in all samples (organs and soil) by SEM-EDS, demonstrating a potential relationship between the neoplasia and the exposure to naturally occurring asbestos (NOA). In conclusion, the results of the present study are further confirmation that wild animals, such as the boar, are suitable sentinels to indicate the risk of environmental exposure to asbestos for human populations.
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Exposição Ambiental/estatística & dados numéricos , Monitoramento Ambiental/métodos , Neoplasias Pulmonares/veterinária , Mesotelioma/veterinária , Animais , Amianto/análise , Asbestose/epidemiologia , Asbestose/veterinária , Poluentes Ambientais/análise , Humanos , Itália , Neoplasias Pulmonares/epidemiologia , Mesotelioma/epidemiologia , Mesotelioma Maligno , SuínosRESUMO
The methods conventionally used to determine the burden of asbestos fibres inhaled/incorporated in lung require chemical digestion of the biological matrix before counting/characterising the inorganic fibrous phases under scanning electron microscopy and energy dispersive spectroscopy (SEM/EDS). Asbestos fibres can also be present in extra-pulmonary organs, and we set out to quantify the fibres in gallbladder. Although the standardised procedure requires approximately 5â¯×â¯10-1â¯g of wet tissue, this amount of tissue is not always available. We applied the procedure on about 9â¯×â¯10-4â¯g of gallbladder from a patient with known environmental and workplace exposure to asbestos. The patient died of malignant pleural mesothelioma and was also affected by severe bile-tract problems. The traditional procedure of digesting tissue samples in NaClO and filtering the resulting suspension was carried out. The filter was then examined under SEM/EDS using two methods 1. following the standardised procedure to assess the fibre burden in lung by investigating only 2â¯mm2 of the filter (660 microscopic fields), and 2. analysing all the microscopic fields in one-quarter of the filter (about 82â¯mm2). In parallel, histological sections (prepared in the usual way for medical diagnosis) were analysed without digestion or manipulation of the sample using variable pressure SEM/EDS. The fibre counts obtained using the two methods were of the same order of magnitude, i.e., â¼105 fibres/g of wet tissue. We showed that the counting of fibres in human tissue may be successfully carried out even when a limited amount of tissue is available. We also found that, when exposure to asbestos is considerable, the number of asbestos fibres accumulating in the gallbladder may be significant.
Assuntos
Asbesto Crocidolita/toxicidade , Asbestos Serpentinas/toxicidade , Vesícula Biliar/química , Neoplasias Pulmonares/mortalidade , Mesotelioma/mortalidade , Exposição Ocupacional/efeitos adversos , Idoso de 80 Anos ou mais , Asbesto Crocidolita/isolamento & purificação , Asbestos Serpentinas/isolamento & purificação , Feminino , Vesícula Biliar/patologia , Humanos , Neoplasias Pulmonares/patologia , Mesotelioma/patologia , Mesotelioma MalignoRESUMO
We studied the effects of fibrous antigorite on mesothelial MeT-5A and monocyte-macrophage J774 cell lines to further understand cellular mechanisms induced by asbestos fibers leading to lung damage and cancer. Antigorite is a mineral with asbestiform properties, which tends to associate with chrysotile or tremolite, and frequently occurs as the predominant mineral in the veins of several serpentinite rocks found abundantly in the Western Alps. Particles containing antigorite are more abundant in the breathing air of this region than those typically found in urban ambient air. Exposure of MeT-5A and J774 cells to fibrous antigorite at concentrations of 5-100 microg/ml for 72 hr induced dose-dependent cytotoxicity. Antigorite also stimulated the ROS production, induced the generation of nitrite and PGE2. MeT-5A cells were more sensitive to antigorite than J774 cells. The results of this study revealed that the fibrous antigorite stimulates cyclooxygenase and formation of hydroxyl and nitric oxide radicals. These changes represent early cellular responses to antigorite fibers, which lead to a host of pathological and neoplastic conditions because free radicals and PGE2 play important roles as mediators of tumor pathogenesis. Understanding the mechanisms of the cellular responses to antigorite and other asbestos particles should be helpful in designing rational prevention and treatment approaches.
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Asbestos Serpentinas/toxicidade , Animais , Testes de Carcinogenicidade , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Dinoprostona/metabolismo , Relação Dose-Resposta a Droga , Humanos , L-Lactato Desidrogenase/metabolismo , Camundongos , Fibras Minerais/toxicidade , Nitritos/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Nitrogênio/metabolismo , Espécies Reativas de Oxigênio/metabolismoRESUMO
Once penetrated into the lungs of exposed people, asbestos induces an in vivo biomineralisation process that leads to the formation of a ferruginous coating embedding the fibres. The ensemble of the fibre and the coating is referred to as asbestos body and is believed to be responsible for the high toxicological outcome of asbestos. Lung tissue of two individuals subjected to prolonged occupational exposure to crocidolite asbestos was investigated using synchrotron radiation micro-probe tools. The distribution of K and of elements heavier than Fe (Zn, Cu, As, and Ba) in the asbestos bodies was observed for the first time. Elemental quantification, also reported for the first time, confirmed that the coating is highly enriched in Fe (~20% w/w), and x-ray absorption spectroscopy indicated that Fe is in the 3+ oxidation state and that it is present in the form of ferritin or hemosiderin. Comparison of the results obtained studying the asbestos bodies upon removing the biological tissue by chemical digestion and those embedded in histological sections, allowed unambiguously distinguishing the composition of the asbestos bodies, and understanding to what extent the digestion procedure altered their chemical composition. A speculative model is proposed to explain the observed distribution of Fe.