Comparison of acute phase response in mice after inhalation and intratracheal instillation of molybdenum disulphide and tungsten particles.

Inhalation studies are the gold standard for assessing the toxicity of airborne materials. They require considerable time, special equipment, and large amounts of test material. Intratracheal instillation is considered a screening and hazard assessment tool as it is simple, quick, allows control of the applied dose, and requires less test material. The particle-induced pulmonary inflammation and acute phase response in mice caused by intratracheal instillation or inhalation of molybdenum disulphide or tungsten particles were compared. End points included neutrophil numbers in bronchoalveolar lavage fluid, Saa3 mRNA levels in lung tissue and Saa1 mRNA levels in liver tissue, and SAA3 plasma protein. Acute phase response was used as a biomarker for the risk of cardiovascular disease. Intratracheal instillation of molybdenum disulphide or tungsten particles did not produce pulmonary inflammation, while molybdenum disulphide particles induced pulmonary acute phase response with both exposure methods and systemic acute phase response after intratracheal instillation. Inhalation and intratracheal instillation showed similar dose-response relationships for pulmonary and systemic acute phase response when molybdenum disulphide was expressed as dosed surface area. Both exposure methods showed similar responses for molybdenum disulphide and tungsten, suggesting that intratracheal instillation can be used for screening particle-induced acute phase response and thereby particle-induced cardiovascular disease.

Genotoxicity in the absence of inflammation after tungsten inhalation in mice.

Tungsten is used in several applications and human exposure may occur. To assess its pulmonary toxicity, we exposed male mice to nose-only inhalation of tungsten particles at 9, 23 or 132 mg/m3 (Low, Mid and High exposure) (45 min/day, 5 days/week for 2 weeks). Increased genotoxicity (assessed by comet assay) was seen in bronchoalveolar (BAL) fluid cells at Low and High exposure. We measured acellular ROS production, and cannot exclude that ROS contributed to the observed genotoxicity. We saw no effects on body weight gain, pulmonary inflammation, lactate dehydrogenase or protein in BAL fluid, pathology of liver or kidney, or on sperm counts. In conclusion, tungsten showed non-dose dependent genotoxicity in the absence of inflammation and therefore interpreted to be primary genotoxicity. Based on genotoxicity, a Lowest Observed Adverse Effect Concentration (LOAEC) could be set at 9 mg/m3. It was not possible to establish a No Adverse Effect Concentration (NOAEC).

Acute phase response following pulmonary exposure to soluble and insoluble metal oxide nanomaterials in mice.

BACKGROUND: Acute phase response (APR) is characterized by a change in concentration of different proteins, including C-reactive protein and serum amyloid A (SAA) that can be linked to both exposure to metal oxide nanomaterials and risk of cardiovascular diseases. In this study, we intratracheally exposed mice to ZnO, CuO, Al2O3, SnO2 and TiO2 and carbon black (Printex 90) nanomaterials with a wide range in phagolysosomal solubility. We subsequently assessed neutrophil numbers, protein and lactate dehydrogenase activity in bronchoalveolar lavage fluid, Saa3 and Saa1 mRNA levels in lung and liver tissue, respectively, and SAA3 and SAA1/2 in plasma. Endpoints were analyzed 1 and 28 days after exposure, including histopathology of lung and liver tissues. RESULTS: All nanomaterials induced pulmonary inflammation after 1 day, and exposure to ZnO, CuO, SnO2, TiO2 and Printex 90 increased Saa3 mRNA levels in lungs and Saa1 mRNA levels in liver. Additionally, CuO, SnO2, TiO2 and Printex 90 increased plasma levels of SAA3 and SAA1/2. Acute phase response was predicted by deposited surface area for insoluble metal oxides, 1 and 28 days post-exposure. CONCLUSION: Soluble and insoluble metal oxides induced dose-dependent APR with different time dependency. Neutrophil influx, Saa3 mRNA levels in lung tissue and plasma SAA3 levels correlated across all studied nanomaterials, suggesting that these endpoints can be used as biomarkers of acute phase response and cardiovascular disease risk following exposure to soluble and insoluble particles.

Pulmonary toxicity of molybdenum disulphide after inhalation in mice.

Molybdenum disulphide (MoS2) is a constituent of many products. To protect humans, it is important to know at what air concentrations it becomes toxic. For this, we tested MoS2 particles by nose-only inhalation in mice. Exposures were set to 13, 50 and 150 mg MoS2/m3 (=8, 30 and 90 mg Mo/m3), corresponding to Low, Mid and High exposure. The duration was 30 min/day, 5 days/week for 3 weeks. Molybdenum lung-deposition levels were estimated based on aerosol particle size distribution measurements, and empirically determined with inductively coupled plasma-mass spectrometry (ICP-MS). Toxicological endpoints were body weight gain, respiratory function, pulmonary inflammation, histopathology, and genotoxicity (comet assay). Acellular reactive oxygen species (ROS) production was also determined. The aerosolised MoS2 powder had a mean aerodynamic diameter of 800 nm, and a specific surface area of 8.96 m2/g. Alveolar deposition of MoS2 in lung was estimated at 7, 27 and 79 µg/mouse and measured as 35, 101 and 171 µg/mouse for Low, Mid and High exposure, respectively. Body weight gain was lower than in controls at Mid and High exposure. The tidal volume was decreased with Low and Mid exposure on day 15. Increased genotoxicity was seen in bronchoalveolar lavage (BAL) fluid cells at Mid and High exposures. ROS production was substantially lower than for carbon black nanoparticles used as bench-mark, when normalised by mass. Yet if ROS of MoS2 was normalised by surface area, it was similar to that of carbon black, suggesting that a ROS contribution to the observed genotoxicity cannot be ruled out. In conclusion, effects on body weight gain and genotoxicity indicated that Low exposure (13 mg MoS2/m3, corresponding to 0.8 mg/m3 for an 8-hour working day) was a No Observed Adverse Effect Concentration (NOAEC,) while effects on respiratory function suggested this level as a Lowest Observed Adverse Effect Concentration (LOAEC).

New method for the protection and restoration of calcareous cultural heritage stones by polyelectrolytes and hydroxyapatite nanocrystals.

We have investigated the feasibility of a new two-step protocol for the restoration of marbles. The process employs a polyelectrolyte multilayer film that enhances the chemical affinity between the treated stone and restorative material (hydroxyapatite nanocrystals), through functionalization, while at the same time it attributes an acid resistant property to the resulting system. Surface functionalization and material deposition is achieved through spraying; a simple and versatile application method suitable for objects of various sizes and geometries. Polyelectrolyte (polyethylenimine and polyacrylic acid) deposition was examined through Attenuated Total Reflection Fourier-Transform Infrared Spectroscopy (ATR-FTIR) and Atomic Force Microscopy (AFM), and tested through contact angle, water absorption and dissolution experiments. The hydroxyapatite nanocrystals were studied by ATR-FTIR, z-potential, AFM and Scanning Electron Microscopy (SEM), and characterized via contact angle and color alteration measurements. Our results show that the polyelectrolyte multilayer was stable in an aqueous environment with increased acid resistance (up to 46% decrease in mass weight loss when compared with untreated samples) and decreased water absorption (up to 39%). Color measurements of the outer hydroxyapatite layer showed a minimal color alteration for one type of the tested substrates showing low color difference values (ΔΕ* < 5). The results suggest that the proposed method holds great potential for marble restoration as it attributes multi-functionality and is easy to apply.

A three-dimensional digital microscopic investigation of entheseal changes as skeletal activity markers.

OBJECTIVES: The current article explores the effectiveness of entheseal changes (EC) as skeletal activity markers by testing the correlation between such changes and cross-sectional geometric (CSG) properties while controlling for the effect of age and body size. MATERIALS AND METHODS: The originality of the article lies in capturing EC in a continuous quantitative manner using three-dimensional microscopy. Roughness and bone resorption were recorded on Zones 1 and 2 of three humeral entheses (subscapularis, supraspinatus, and infraspinatus) in a documented sample of 29 male skeletons. RESULTS: Our analysis found that merely 5.91% of the partial correlations between EC and CSG properties were statistically significant. In addition, two unexpected patterns were identified, namely a higher number of significant correlations on the left side entheses compared to the right side ones, and a higher number of correlations between minimum roughness and CSG properties compared to mean and maximum roughness. DISCUSSION: These patterns are the inverse of what we would expect if activity had exerted an important effect on EC expression. Therefore, they support the lack of association between EC and habitual activity, even though various factors potentially affecting the above results are discussed.

Sand consolidation with calcium phosphate-polyelectrolyte composites.

A new method for the consolidation of loose sand formations has been developed. The method involves in situ precipitation of a composite calcium phosphate-polyelectrolyte salt that binds together with loose sand grains, thus resulting to their consolidation. Three different polyelectrolytes (PE) were tested, i.e., polyacrylic acid (PAA), polyallylamine hydrochloride (PAH), and polyethylenimine (PEI). The effect of PE tested on the thermodynamics and the kinetics of precipitation of calcium phosphate salts was investigated. Three types of experiments were done. Investigation of the adsorption of PE on either hydroxyapatite (Ca(5)(PO(4))(3)OH, HAP) crystals or on sand grains. Measurement of the kinetics of heterogeneous nucleation of HAP on the solid substrates and the mechanical properties of the obtained crystals in batch experiments of low and high supersaturation solutions, respectively. Evaluation of the consolidation in sand packs in order to investigate the effectiveness of the method. The crystallization rates, R(p), on HAP crystals in the presence of the PE tested were found in the order R(p)(PAA)>R(p)(PEI)>R(p)(PAH), while nucleation and crystal growth on silicate sand took place only in the absence of adsorbed PE. PAH favored strongly the consolidation process, whereas PEI and PAA resulted in the formation of poorly consolidated grain agglomerates.