William P. Chisholm

Multiwalled Carbon Nanotubes Induce a Fibrogenic Response by Stimulating Reactive Oxygen Species Production, Activating NF-κB Signaling, and Promoting Fibroblast-to-Myofibroblast Transformation

Carbon nanotubes (CNTs) are novel materials with unique electronic and mechanical properties. The extremely small size, fiberlike shape, large surface area, and unique surface chemistry render their distinctive chemical and physical characteristics and raise potential hazards to humans. Several reports have shown that pulmonary exposure to CNTs caused inflammation and lung fibrosis in rodents. The molecular mechanisms that govern CNT lung toxicity remain largely unaddressed. Here, we report that multiwalled carbon nanotubes (MWCNTs) have potent, dose-dependent toxicity on cultured human lung cells (BEAS-2B, A549, and WI38-VA13). Mechanistic analyses were carried out at subtoxic doses (≤20 μg/mL, ≤ 24 h). MWCNTs induced substantial ROS production and mitochondrial damage, implicating oxidative stress in cellular damage by MWCNT. MWCNTs activated the NF-κB signaling pathway in macrophages (RAW264.7) to increase the secretion of a panel of cytokines and chemokines (TNFα, IL-1β, IL-6, IL-10, and MCP1) that promote inflammation. Activation of NF-κB involved rapid degradation of IκBα, nuclear accumulation of NF-κBp65, binding of NF-κB to specific DNA-binding sequences, and transactivation of target gene promoters. Finally, MWCNTs induced the production of profibrogenic growth factors TGFβ1 and PDGF from macrophages that function as paracrine signals to promote the transformation of lung fibroblasts (WI38-VA13) into myofibroblasts, a key step in the development of fibrosis. Our results revealed that MWCNTs elicit multiple and intertwining signaling events involving oxidative damage, inflammatory cytokine production, and myofibroblast transformation, which potentially underlie the toxicity and fibrosis in human lungs by MWCNTs.

Performance of High Flow Rate Samplers for Respirable Particle Collection

The American Conference of Governmental Industrial hygienists (ACGIH) lowered the threshold limit value (TLV) for respirable crystalline silica (RCS) exposure from 0.05 to 0.025 mg m−3 in 2006. For a working environment with an airborne dust concentration near this lowered TLV, the sample collected with current standard respirable aerosol samplers might not provide enough RCS for quantitative analysis. Adopting high flow rate sampling devices for respirable dust containing silica may provide a sufficient amount of RCS to be above the limit of quantification even for samples collected for less than full shift. The performances of three high flow rate respirable samplers (CIP10-R, GK2.69, and FSP10) have been evaluated in this study. Eleven different sizes of monodisperse aerosols of ammonium fluorescein were generated with a vibrating orifice aerosol generator in a calm air chamber in order to determine the sampling efficiency of each sampler. Aluminum oxide particles generated by a fluidized bed aerosol generator were used to test (i) the uniformity of a modified calm air chamber, (ii) the effect of loading on the sampling efficiency, and (iii) the performance of dust collection compared to lower flow rate cyclones in common use in the USA (10-mm nylon and Higgins–Dewell cyclones). The coefficient of variation for eight simultaneous samples in the modified calm air chamber ranged from 1.9 to 6.1% for triplicate measures of three different aerosols. The 50% cutoff size (50dae) of the high flow rate samplers operated at the flow rates recommended by manufacturers were determined as 4.7, 4.1, and 4.8 μm for CIP10-R, GK2.69, and FSP10, respectively. The mass concentration ratio of the high flow rate samplers to the low flow rate cyclones decreased with decreasing mass median aerodynamic diameter (MMAD) and high flow rate samplers collected more dust than low flow rate samplers by a range of 2–11 times based on gravimetric analysis. Dust loading inside the high flow rate samplers does not appear to affect the particle separation in either FSP10 or GK2.69. The high flow rate samplers overestimated compared to the International Standards Organization/Comité Européen de Normalisation/ACGIH respirable convention [up to 40% at large MMAD (27.5 μm)] and could provide overestimated exposure data with the current flow rates. However, both cyclones appeared to be able to provide relatively unbiased assessments of RCS when their flow rates were adjusted.

Quartz Measurement in Coal Dust with High-Flow Rate Samplers: Laboratory Study

A laboratory study was performed to measure quartz in coal dust using high-flow rate samplers (CIP10-R, GK2.69 cyclone, and FSP10 cyclone) and low-flow rate samplers [10-mm nylon and Higgins-Dewell type (BGI4L) cyclones] and to determine whether an increased mass collection from high-flow rate samplers would affect the subsequent quartz measurement by Fourier transform infrared (FTIR) and X-ray diffraction (XRD) analytical procedures. Two different sizes of coal dusts, mass median aerodynamic diameter 4.48 μm (Coal Dust A) and 2.33 μm (Coal Dust B), were aerosolized in a calm air chamber. The mass of coal dust collected by the samplers was measured gravimetrically, while the mass of quartz collected by the samplers was determined by FTIR (NIOSH Manual of Analytical Method 7603) and XRD (NIOSH Manual of Analytical Method 7500) after one of two different indirect preparations. Comparisons between high-flow rate samplers and low-flow rate samplers were made by calculating mass concentration ratios of coal dusts, net mass ratios of coal dusts, and quartz net mass. Mass concentrations of coal dust from the FSP10 cyclone were significantly higher than those from other samplers and mass concentrations of coal dust from 10-mm nylon cyclone were significantly lower than those from other samplers, while the CIP10-R, GK2.69, and BGI4L samplers did not show significant difference in the comparison of mass concentration of coal dusts. The BGI4L cyclone showed larger mass concentration of ∼9% compared to the 10-mm nylon cyclone. All cyclones provided dust mass concentrations that can be used in complying with the International Standard Organization standard for the determination of respirable dust concentration. The amount of coal dust collected from the high-flow rate samplers was found to be higher with a factor of 2-8 compared to the low-flow rate samplers but not in direct proportion of increased flow rates. The high-flow rate samplers collected more quartz compared to low-flow rate samplers in the range of 2-10. There was no significant difference between the per cent (%) quartz in coal dust between the FTIR and XRD analyses. The findings of this study indicated that the increased mass of quartz collected with high-flow rate samplers would provide precise analytical results (i.e. significantly above the limit of detection and/or limit of quantification) compared to the mass collected with low-flow rate samplers, especially in environments with low concentrations of quartz or where short sampling times are desired.

Toxicological Assessment of CoO and La2O3 Metal Oxide Nanoparticles in Human Small Airway Epithelial Cells

Cobalt monoxide (CoO) and lanthanum oxide (La2O3) nanoparticles are 2 metal oxide nanoparticles with different redox potentials according to their semiconductor properties. By utilizing these two nanoparticles, this study sought to determine how metal oxide nanoparticles mode of toxicological action is related to their physio-chemical properties in human small airway epithelial cells (SAEC). We investigated cellular toxicity, production of superoxide radicals and alterations in gene expression related to oxidative stress, and cellular death at 6 and 24 h following exposure to CoO and La2O3(administered doses: 0, 5, 25, and 50 µg/ml) nanoparticles. CoO nanoparticles induced gene expression related to oxidative stress at 6 h. After characterizing the nanoparticles, transmission electron microscope analysis showed SAEC engulfed CoO and La2O3nanoparticles. CoO nanoparticles were toxic after 6 and 24 h of exposure to 25.0 and 50.0 µg/ml administered doses, whereas, La2O3nanoparticles were toxic only after 24 h using the same administered doses. Based upon the Volumetric Centrifugation Methodin vivoSedimentation, Diffusion, and Dosimetry, the dose of CoO and La2O3nanoparticles delivered at 6 and 24 h were determined to be: CoO: 1.25, 6.25, and 12.5 µg/ml; La2O3: 5, 25, and 50 µg/ml and CoO: 4, 20, and 40 µg/ml; and La2O3: 5, 25, 50 µg/ml, respectively. CoO nanoparticles produced more superoxide radicals and caused greater stimulation of total tyrosine and threonine phosphorylation at both 6 and 24 h when compared with La2O3nanoparticles. Taken together, these data provide evidence that different toxicological modes of action were involved in CoO and La2O3metal oxide nanoparticle-induced cellular toxicity.

A Field Study on the Respiratory Deposition of the Nano-Sized Fraction of Mild and Stainless Steel Welding Fume Metals.

A field study was conducted to estimate the amount of Cr, Mn, and Ni deposited in the respiratory system of 44 welders in two facilities. Each worker wore a nanoparticle respiratory deposition (NRD) sampler during gas metal arc welding (GMAW) of mild and stainless steel and flux-cored arc welding (FCAW) of mild steel. Several welders also wore side-by-side NRD samplers and closed-face filter cassettes for total particulate samples. The NRD sampler estimates the aerosols nano-fraction deposited in the respiratory system. Mn concentrations for both welding processes ranged 2.8–199 μg/m3; Ni concentrations ranged 10–51 μg/m3; and Cr concentrations ranged 40–105 μg/m3. Cr(VI) concentrations ranged between 0.5–1.3 μg/m3. For the FCAW process the largest concentrations were reported for welders working in pairs. As a consequence this often resulted in workers being exposed to their own welding fumes and to those generated from the welding partner. Overall no correlation was found between air velocity and exposure (R2 = 0.002). The estimated percentage of the nano-fraction of Mn deposited in a mild-steel-welders respiratory system ranged between 10 and 56%. For stainless steel welding, the NRD samplers collected 59% of the total Mn, 90% of the total Cr, and 64% of the total Ni. These results indicate that most of the Cr and more than half of the Ni and Mn in the fumes were in the fraction smaller than 300 nm.

Size Distributions of 0.5 to 20 μm Aerodynamic Diameter Lead-Containing Particles from Aerosol Sampler Walls and Filters

The study presented here investigates the number weighted particle size distributions of aerosols generated in the laboratory from lead oxide and lead sulfide dusts and sampled by Institute of Occupational Medicine (IOM) and closed face cassette (CFC) samplers as determined by scanning electron microscopy (SEM). The wall deposits and filter deposits from each sampler were characterized separately. A Mann-Whitney statistical analysis revealed that differences in the number weighted distributions of particles captured by the filter and the wall were not significant over the size range (up to 20 μm aerodynamic equivalent diameter) present in these laboratory-generated aerosols. Furthermore, for these samples it was not possible to distinguish an absolute difference between the IOM and CFC filter catches. By comparing direct measurements of aerodynamic equivalent diameter (AED) made by an Aerodynamic Particle Sizer (APS) to AEDs calculated from SEM images, empirical shape factors for lead oxide and lead sulfide were determined. To validate this approach APS and SEM measurements of the AED of 2 μm and 6 μm physical diameter monodisperse glass and polystyrene microspheres were made. Using the shape factors of spheres and the known densities of these materials, it was found that the SEM determinations of AED agreed with the APS results. To demonstrate the reliability of the redeposition method of sample preparation, lead sulfide and lead oxide aerosols were briefly sampled by IOM samplers such that sufficient particles were collected for SEM examination directly on the filter but not so many that particles were likely to touch or overlap. Half of each filter was analyzed in the SEM directly; the other half was ultrasonically removed and re-deposited for analysis by SEM. There were no statistically significant differences in their number weighted size distributions, demonstrating that the sample treatment process does not change the size distribution of these particular aerosols.

Comparison of Filter and Wall Deposits From Samplers Used to Collect Airborne Lead-Containing Dusts at Field Sites

Pairs of Institute of Occupational Medicine (IOM) and 37 mm closed face cassette samplers (CFC) were deployed where occupational exposures to lead-containing dusts were known to occur. Discrete particle analyses of wall and filter deposits were performed by Scanning Electron Microscopy—Energy Dispersive X-ray Spectrometry (SEM-EDX). From the elemental composition and projected area diameter of each particle a density, volume, and mass were calculated, and a mass-weighted size distribution for each filter and corresponding wall deposit determined. Comparison of pairs of wall and filter mass-weighted size distributions by Mann–Whitney statistical analysis shows that in only 3 of 72 examples from either sampler were the distributions significantly different that suggests that the mechanisms of particle deposition on the sampler walls for particles in this size range (0.5 μm through 20 μm) do not differ for the different samplers. Furthermore, in only 4 of 33 sampler pairs did the IOM and CFC results differ. Although these results originate from several distinct processes characterized by different chemical and physical dust generation mechanisms, they suggest that in these environments the measurement of “total dust” by the CFC and inhalable dust by the IOM will be very similar when both samplers are processed the same way with respect to the including or excluding wall deposits with the filter catch. However, these results may not be applicable to environments where larger particles exist.

Size Distribution and Estimated Respiratory Deposition of Total Chromium, Hexavalent Chromium, Manganese, and Nickel in Gas Metal Arc Welding Fume Aerosols

A laboratory study was conducted to determine the mass of total Cr, Cr(VI), Mn, and Ni in 15 size fractions for mild and stainless steel gas-metal arc welding (GMAW) fumes. Samples were collected using a nano multi orifice uniform deposition impactor (MOUDI) with polyvinyl chloride filters on each stage. The filters were analyzed by inductively coupled plasma mass spectrometry (ICP-MS) and ion chromatography. Limits of detection (LODs) and quantitation (LOQs) were experimentally calculated and percent recoveries were measured from spiked metals in solution and dry, certified welding-fume reference material. The fraction of Cr(VI) in total Cr was estimated by calculating the ratio of Cr(VI) to total Cr mass for each particle size range. Expected, regional deposition of each metal was estimated according to respiratory-deposition models. The weight percent (standard deviation) of Mn in mild steel fumes was 9.2% (6.8%). For stainless steel fumes, the weight percentages were 8.4% (5.4%) for total Cr, 12.2% (6.5%) for Mn, 2.1% (1.5%) for Ni and 0.5% (0.4%) for Cr(VI). All metals presented a fraction between 0.04 and 0.6 μm. Total Cr and Ni presented an additional fraction <0.03 μm. On average 6% of the Cr was found in the Cr(VI) valence state. There was no statistical difference between the smallest and largest mean Cr(VI) to total Cr mass ratio (p-value = 0.19), hence our analysis does not show that particle size affects the contribution of Cr(VI) to total Cr. The predicted total respiratory deposition for the metal particles was ∼25%. The sites of principal deposition were the head airways (7–10%) and the alveolar region (11–14%). Estimated Cr(VI) deposition was highest in the alveolar region (14%).

Replacement of filters for respirable quartz measurement in coal mine dust by infrared spectroscopy

ABSTRACT The objective of this article is to compare and characterize nylon, polypropylene (PP), and polyvinyl chloride (PVC) membrane filters that might be used to replace the vinyl/acrylic co-polymer (DM-450) filter currently used in the Mine Safety and Health Administration (MSHA) P-7 method (Quartz Analytical Method) and the National Institute for Occupational Safety and Health (NIOSH) Manual of Analytical Methods 7603 method (QUARTZ in coal mine dust, by IR re-deposition). This effort is necessary because the DM-450 filters are no longer commercially available. There is an impending shortage of DM-450 filters. For example, the MSHA Pittsburgh laboratory alone analyzes annually approximately 15,000 samples according to the MSHA P-7 method that requires DM-450 filters. Membrane filters suitable for on-filter analysis should have high infrared (IR) transmittance in the spectral region 600–1000 cm−1. Nylon (47 mm, 0.45 µm pore size), PP (47 mm, 0.45 µm pore size), and PVC (47 mm, 5 µm pore size) filters meet this specification. Limits of detection and limits of quantification were determined from Fourier transform infrared spectroscopy (FTIR) measurements of blank filters. The average measured quartz mass and coefficient of variation were determined from test filters spiked with respirable α-quartz following MSHA P-7 and NIOSH 7603 methods. Quartz was also quantified in samples of respirable coal dust on each test filter type using the MSHA and NIOSH analysis methods. The results indicate that PP and PVC filters may replace the DM-450 filters for quartz measurement in coal dust by FTIR. PVC filters of 5 µm pore size seemed to be suitable replacement although their ability to retain small particulates should be checked by further experiment.

Comparison of Lead and Tin Concentrations in Air at a Solder Manufacturer from the Closed-Face 37-mm Cassette With and Without a Custom Cellulose-Acetate Cassette Insert

A polyvinyl chloride (PVC) cassette insert with PVC filter (ACCU-CAP) in a 37-mm closed-face cassette (CFC) was designed for gravimetric analysis. A customized version of the ACCU-CAP, also to be used in the CFC, was manufactured from an acid-digestible cellulose-acetate cassette insert joined to a mixed cellulose ester (MCE) filter for wet chemical analysis. The aim of this study was to compare metal particle concentrations as sampled by the customized insert (CI) in a CFC sampler with the traditional sampling method using only a MCE filter in the CFC. Thirty-nine personal and 13 area samples were taken using paired filter-based CFC and the CI in CFC samplers at a solder manufacturing plant. The CI was removed from its CFC, and digested and analyzed as a whole. The MCE filter from the typical CFC was removed for analysis and then the interior of the cassette was wiped with Ghost Wipe for a separate analysis. The MCE filter only, Ghost Wipe, and CI were separately dissolved in heated nitric acid for ICP-MS analysis. Overall, the geometric mean concentration of the filter-only (FO) samples was considerably lower than that of the CI samples, by 53% for lead and 32% for tin. However, if the FO analysis was added to the corresponding Ghost Wipe analysis, i.e., filter+interior wipe (FW), the geometric mean concentrations of the FW results were similar to those of the CI results (by 113% for lead and 98% for tin). For both lead and tin the comparison of (log-transformed) metal concentrations between the FW and CI results showed no statistically significant difference (p-value = 0.3009 for lead and 0.800 for tin), while the comparison between the FO and CI results shows statistically significant differences (all p-values < 0.05). In conclusion, incorporating the sampler internal non-filter deposits by wiping or use of an internal filter capsule gave higher results than analyzing only the filter. Close agreement between the two methods of including non-filter deposits is an indication of general equivalency.