Dissolution Reaction and Surface Modification of UICC Amosite in Mimicked Gamble's Solution: A Step towards Filling the Gap between Asbestos Toxicity and Its Crystal Chemical Features.

This study focuses on the dissolution process and surface characterization of amosite fibres following interaction with a mimicked Gamble's solution at a pH of 4.5 and T = 37 °C, up to 720 h. To achieve this, a multi-analytical approach was adopted, and the results were compared to those previously obtained on a sample of asbestos tremolite and UICC crocidolite, which were investigated under the same experimental conditions. Combining surface chemical data obtained by XPS with cation release quantified by ICP-OES, an incongruent behaviour of the fibre dissolution was highlighted for amosite fibres, similarly to asbestos tremolite and UICC crocidolite. In particular, a preferential release of Mg and Ca from the amphibole structure was observed, in agreement with their Madelung site energies. Notably, no Fe release from amosite fibres was detected in our experimental conditions (pH of 4.5 and atmospheric pO2), despite the occurrence of Fe(II) at the M(4) site of the amphibole structure, where cations are expected to be rapidly leached out during mineral dissolution. Moreover, the oxidation of both the Fe centres initially present on the fibre surface and those promoted from the bulk, because of the erosion of the outmost layers, was observed. Since biodurability (i.e., the resistance to dissolution) is one of the most important toxicity parameters, the knowledge of the surface alteration of asbestos possibly occurring in vivo may help to understand the mechanisms at the basis of its long-term toxicity.

Particle Debris Generated from Passenger Tires Induces Morphological and Gene Expression Alterations in the Macrophages Cell Line RAW 264.7.

Air pollution in the urban environment is a topical subject. Aero-suspended particles can cause respiratory diseases in humans, ranging from inflammation to asthma and cancer. One of the components that is most prevalent in particulate matter (PM) in urban areas is the set of tire microparticles (1-20 µm) and nanoparticles (<1 µm) that are formed due to the friction of wheels with asphalt and are increased in slow-moving areas that involve a lot of braking actions. In this work, we studied the effect that microparticles generated from passenger tires (PTWP, passenger tire wear particles) have in vitro on murine macrophages cells RAW 264.7 at two concentrations of 25 and 100 µg/mL, for 24 and 48 h. In addition to the chemical characterization of the material and morphological characterization of the treated cells by transmission electron microscopy, gene expression analysis with RT-PCR and active protein analysis with Western blotting were performed. Growth curves were obtained, and the genotoxic effect was evaluated with a comet assay. The results indicate that initially, an induction of the apoptotic process is observable, but this is subsequently reversed by Bcl2. No genotoxic damage is present, but mild cellular abnormalities were observed in the treated cells.

Multidisciplinary investigation of an ancient renal stone in a mummy from Popoli, central Italy.

The renal stone found in the natural mummy of an anonymous nobleman dating to 19th century was investigated using advanced imaging modalities and analytic investigations. By this multidisciplinary approach we were able to identify the chemical components and their distribution throughout the sample. These results allowed to understand the lifestyle habits of the subject, as well as the exact pathogenesis of his disease.

Use of Lichens to Evaluate the Impact of Post-Earthquake Reconstruction Activities on Air Quality: A Case Study from the City of L'Aquila.

Lichens are widely used as bioindicators of air quality because of their ability to absorb chemical pollutants. We used the Lichen Diversity Value (LDV) index to assess the effects of the urban reconstruction activities in the city of L'Aquila ten years after the 2009 earthquake on air quality. Sampling was conducted from the city centre (still mostly under reconstruction and closed to traffic) to suburban areas (where reconstruction is minimal). We tested if the LDV index varied with distance from the city centre because of the presence of air pollutants produced by reconstruction works. We also used Energy-Dispersive X-ray Spectroscopy (EDS) to detect the main pollutants accumulated in the sampled lichens. The LDV increased from the city centre towards suburban areas. EDS revealed high concentrations of pollutants related to demolition and reconstruction activities, such as aluminium and silicon (used in the manufacture of concrete), in the more central areas. These results suggest that the LDV index can be a useful tool to monitor air quality, even on a small scale, and in urban environments subject to building demolition and reconstruction. Moreover, EDS could represent a good preliminary analytical technique to identify the air pollutants associated with all of these activities.

Genotoxicity Response of Fibroblast Cells and Human Epithelial Adenocarcinoma In Vitro Model Exposed to Bare and Ozone-Treated Silica Microparticles.

Indoor air pollutants (IAP), which can pose a serious risk to human health, include biological pollutants, nitric oxide (NO), nitrogen dioxide (NO2), volatile organic compounds (VOC), sulfur dioxide (SO2), carbon monoxide (CO), carbon dioxide (CO2), silica, metals, radon, and particulate matter (PM). The aim of our work is to conduct a multidisciplinary study of fine silica particles (<2.5 µm) in the presence or absence of ozone (O3), and evaluate their potential cytotoxicity using MTS, micronucleus, and the comet test in two cell lines. We analyzed A549 (human basal alveolar epithelial cell adenocarcinoma) and Hs27 (human normal fibroblasts) exposed to dynamic conditions by an IRC simulator under ozone flow (120 ppb) and in the presence of silica particles (40 µg/h). The viability of A549 and Hs27 cells at 48 and 72 h of exposure to silica or silica/ozone decreases, except at 72 h in Hs27 treated with silica/ozone. The micronucleus and comet tests showed a significant increase in the number of micronuclei and the % of DNA in the queue, compared to the control, in both lines in all treatments, even if in different cell times/types. We found that silica alone or with more O3 causes more pronounced genotoxic effects in A549 tumor cells than in normal Hs27 fibroblasts.

Surface and bulk modifications of amphibole asbestos in mimicked gamble's solution at acidic PH.

This study aimed at investigating the surface modifications occurring on amphibole asbestos (crocidolite and tremolite) during leaching in a mimicked Gamble's solution at pH of 4.5 and T = 37 °C, from 1 h up to 720 h. Results showed that the fibre dissolution starts with the release of cations prevalently allocated at the various M- and (eventually) A-sites of the amphibole structure (incongruent dissolution). The amount of released silicon, normalized to fibre surface area, highlighted a leaching faster for the crocidolite sample, about twenty times higher than that of tremolite. Besides, the fast alteration of crocidolite promotes the occurrence of Fe centres in proximity of the fibre surface, or possibly even exposed, particularly in the form of Fe(II), of which the bulk is enriched with respect to the oxidized surface. Conversely, for tremolite fibres the very slow fibre dissolution prevents the underlying cations of the bulk to be exposed on the mineral surface, and the iron oxidation, faster than the leaching process, significantly depletes the surface Fe(II) centres initially present. Results of this work may contribute to unravel possible correlations between surface properties of amphibole asbestos and its long-term toxicity.

In Vitro Shear Bond Strength of Orthodontic Brackets after Enamel Conditioning with Acid Etching and Hydroabrasion.

The purpose of this study was to evaluate the shear bond strength and adhesive remnant index ARI) of orthodontic brackets following enamel conditioning with acid etching, hydroabrasion, and with both procedures. Thirty extracted human premolars were divided into three groups and received either acid etching, hydroabrasion or both procedures. Orthodontic brackets were bonded with composite resin. Shear bond strength was tested with a tensile machine, then the teeth were observed under a stereomicroscope to evaluate ARI scores. The enamel morphology after each conditioning method was evaluated with scanning electron microscope imaging. A one-way ANOVA and a Kruskal-Wallis H test were used to compare the bond strength and the ARI scores among the three groups. Hydroabrasion alone produced shear bond strength values below clinical acceptability, while the combination of acid etching and hydroabrasion produced the highest values. The ARI scores in the hydroabrasion group were significantly different from the other groups. Hydroabrasion followed by acid etching was effective in increasing the shear bond strength of orthodontic brackets. Further in vivo studies are needed to confirm the cost and benefits of this technique.

Thermal inertization of amphibole asbestos modulates Fe topochemistry and surface reactivity.

This study discloses the morphological and chemical-structural modifications that occur during thermal degradation of amphibole asbestos. Low-iron tremolite and iron-rich crocidolite were heated at temperatures ranging from r.t. to 1200 °C. Heating promoted a complex sequence of iron oxidation, migration and/or clustering and, finally, the formation of brittle fibrous pseudomorphs consisting of newly formed minerals and amorphous nanophases. The effects of the thermal modifications on toxicologically relevant asbestos reactivity were evaluated by quantifying carbon- and oxygen-centred, namely hydroxyl (OH), radicals. Heating did not alter carbon radicals, but largely affected oxygen-centred radical yields. At low temperature, reactivity of both amphiboles decreased. At 1200 °C, tremolite structural breakdown was achieved and the reactivity was further reduced by migration of reactive iron ions into the more stable TO4 tetrahedra of the newly formed pyroxene(s). Differently, crocidolite breakdown at 1000 °C induced the formation of hematite, Fe-rich pyroxene, cristobalite, and abundant amorphous material and restored radical reactivity. Our finding suggests that thermally treated asbestos and its breakdown products still share some toxicologically relevant properties with pristine fibre. Asbestos inertization studies should consider morphology and surface reactivity, beyond crystallinity, when proving that a thermally inactivated asbestos-containing material is safe.

Enamel preservation during composite removal after orthodontic debonding comparing hydroabrasion with rotary instruments.

The aim of the present study was to evaluate how hydroabrasion performs during composite removal. A standardized amount of composite was bonded to 40 enamel surfaces of extracted third molars, then removed with either a tungsten carbide bur mounted on a micro-motor handpiece without irrigation, a tungsten carbide bur mounted on a micro-motor handpiece with irrigation, a tungsten carbide bur mounted on an air-rotor handpiece, or hydroabrasion. The four treatment methods were compared using the enamel-surface-index and the adhesive-remnant-index and performing a Kruskal-Wallis statistical test to detect differences between each method' scores. Hydroabrasion produced significantly less damage to the enamel surface compared to the other three methods. Hydroabrasion was the cleaning method that produced less damages to the enamel surface, at a cost of a less efficient composite removal than tungsten carbide burs on micro-motor handpiece.

In Vitro Genotoxicity of Polystyrene Nanoparticles on the Human Fibroblast Hs27 Cell Line.

Several studies have provided information on environmental nanoplastic particles/debris, but the in vitro cyto-genotoxicity is still insufficiently characterized. The aim of this study is to analyze the effects of polystyrene nanoparticles (PNPs) in the Hs27 cell line. The viability of Hs27 cells was determined following exposure at different time windows and PNP concentrations. The genotoxic effects of the PNPs were evaluated by the cytokinesis-block micronucleus (CBMN) assay after exposure to PNPs. We performed ROS analysis on HS27 cells to detect reactive oxygen species at different times and treatments in the presence of PNPs alone and PNPs added to the Crocus sativus L. extract. The different parameters of the CBMN test showed DNA damage, resulting in the increased formation of micronuclei and nuclear buds. We noted a greater increase in ROS production in the short treatment times, in contrast, PNPs added to Crocus sativus showed the ability to extract, thus reducing ROS production. Finally, the SEM-EDX analysis showed a three-dimensional structure of the PNPs with an elemental composition given by C and O. This work defines PNP toxicity resulting in DNA damage and underlines the emerging problem of polystyrene nanoparticles, which extends transversely from the environment to humans; further studies are needed to clarify the internalization process.

Exposure to particle debris generated from passenger and truck tires induces different genotoxicity and inflammatory responses in the RAW 264.7 cell line.

A number of studies have shown variable grades of cytotoxicity and genotoxicity in in vitro cell cultures, laboratory animals and humans when directly exposed to particle debris generated from tires. However, no study has compared the effects of particles generated from passenger tires with the effects of particles from truck tires. The aim of this study was to investigate and relate the cyto- and genotoxic effects of different types of particles (PP, passenger tire particles vs. TP, truck tire particles) in vitro using the phagocytic cell line RAW 264.7 (mouse leukaemic monocyte macrophage cell line). The viability of RAW 264.7 cells was determined by the 3- (4,5-dimethylthiazol-2-yl) -5- (3-carboxymethoxyphenyl) -2- (4-sulfophenyl) -2H-tetrazolium (MTS) assay following exposure for 4, 24 and 48 hours to different particle concentrations (10 µg / ml, 25 µg / ml, 50 µg / ml, 100 µg / ml). The effects of particles of passenger and truck tires on cell proliferation and genotoxicity were evaluated by means of the cytokinesis-block micronucleus (CBMN) assay following exposure for 24 hours to different particle concentrations (10 µg / ml, 25 µg / ml, 50 µg / ml, 100 µg / ml). In MTS assay, after 24 hours, it was found that PP induced a 30% decrease in metabolic activity at a concentration of 10 µg/ml, while TP caused reductions of 20% and 10% at concentrations of 10 µg/ml and 50 µg/ml, respectively. At 48 hours after the treatments, we observed increased metabolic activity at 50 µg/ml and 100 µg/ml for the PP while only at 50 µg/ml for the TP. The CBMN assay showed a significant increase in the number of micronuclei in the cells incubated with PP in all experimental conditions, while the cells treated with TP showed a meaningful increase only at 10 µg /ml. We utilized the TNF-α ELISA mouse test to detect the production of tumour necrosis factor-alpha (TNF-α) in RAW 264.7 cells. The effect of passenger and truck particles on TNF-α release was evaluated following exposure for 4 and 24 hours. After 4 hours of incubation, the cells treated with PP and TP at 100 µg / ml showed a slight but significant increase in TNF-α release, while there was a significant increase in the release of TNF-α after 24 hours of incubation with both tire samples in the cells treated with 50 and 100 µg / ml PP. The data obtained show a higher cytotoxic, clastogenic/genotoxic and inflammatory effects of passenger compared to the truck tire particles.

Characterization of atmospheric particulate: relationship between chemical composition, size, and emission source.

Physicochemical characterization of particulate matter fractionated into eight samples by size from 10 to 0.43 microm was performed by HS-SPME/GC-MS for the organic (semi-)volatile components and SEM X-ray microanalysis (SEM/EDX) for analysis of the elemental composition. The HS-SPME technique was shown to be efficient with respect to requiring an extremely low amount of material, being selective and clean and avoiding use of any solvents. Particulate matter was collected at four sites characterized by particular environmental locations and different pollution levels around the city of L'Aquila in central Italy. The results reveal a tight correlation between the particle composition, size, and the emission source. The analyses show also that the finer the particle, the higher its content of elemental carbon and organic compounds. Well-known carcinogens such as PAHs were detected among the identified organic compounds from both the rural and highly polluted sites.

[Inhalable airborne particulate pollution in sites characterized by dissimilar environmental conditions]. / Inquinamento da polveri e da particolato fino in siti con differenti caratteristiche ambientali.

The aim of the present study is to characterise from a physico-chemical point of view the airborne particulate matter (PM10) sampled in different sites dissimilar with regard to the weather conditions, the residential density and the industrial activities. The particles were collected by an eight-stage cascade impactor, Andersen particle fractionating sampler. The analysis of the particle samples was performed by a scanning electron microscopy (SEM) equipped with a thin-window system for X-ray microanalysis by energy dispersion spectrometry. The hierarchical cluster analysis (HCA) on the analytical data revealed the presence of eight different particulate types (particle clusters) in the sampling sites: C-rich particles (cluster 1); Ca and Mg carbonates (cluster 2); Ca sulphates (cluster 3); silica particles (cluster 4); silicates (cluster 5); Fe-rich particles (cluster 6); metal compounds (cluster 7); carbonaceous particles with heavy metals (cluster 8). Data obtained in this study demonstrate a significant correlation between the PM10 composition and the anthropic activities present the sampling site.

Monitoring urban air particulate matter (fractions PM 2.5 and PM 10) genotoxicity by plant systems and human cells in vitro: a comparative analysis.

Increased incidence of mortality and sickness due to cardiopulmonary complications has been associated with elevated levels of urban air particles (UAP), with an aerodynamic diameter of 10 microm (PM 10) and 2.5 microm (PM 2.5). In the present report alternative plant systems and human cells in vitro are associated with human hazard and genotoxic risk assessment of UAP. The genotoxic activities associated with the coarse (PM 10) and the fine fraction (PM 2.5) of airborne particulates have been analyzed by evaluating micronuclei induction and/or sister-chromatid exchange (SCE) using in vitro models of Daucus carota and HS 27 human fibroblast cell suspensions and Zea mays root meristems. Results show variability in the response of the test systems and indicate that the mutagenicity trend in both plant and human cell cultures was directly correlated to the concentration of carbon-rich particles in the fraction of the PM 2.5 airborne particulates. Moreover, in plant tissues, the frequency of micronuclei and SCE was related to an enhancement of the specific activity of the stress-related enzyme peroxidase.