Histopathology of the broad class of carbon nanotubes and nanofibers used or produced in U.S. facilities in a murine model.

BACKGROUND: Multi-walled carbon nanotubes and nanofibers (CNT/F) have been previously investigated for their potential toxicities; however, comparative studies of the broad material class are lacking, especially those with a larger diameter. Additionally, computational modeling correlating physicochemical characteristics and toxicity outcomes have been infrequently employed, and it is unclear if all CNT/F confer similar toxicity, including histopathology changes such as pulmonary fibrosis. Male C57BL/6 mice were exposed to 40 µg of one of nine CNT/F (MW #1-7 and CNF #1-2) commonly found in exposure assessment studies of U.S. facilities with diameters ranging from 6 to 150 nm. Human fibroblasts (0-20 µg/ml) were used to assess the predictive value of in vitro to in vivo modeling systems. RESULTS: All materials induced histopathology changes, although the types and magnitude of the changes varied. In general, the larger diameter MWs (MW #5-7, including Mitsui-7) and CNF #1 induced greater histopathology changes compared to MW #1 and #3 while MW #4 and CNF #2 were intermediate in effect. Differences in individual alveolar or bronchiolar outcomes and severity correlated with physical dimensions and how the materials agglomerated. Human fibroblast monocultures were found to be insufficient to fully replicate in vivo fibrosis outcomes suggesting in vitro predictive potential depends upon more advanced cell culture in vitro models. Pleural penetrations were observed more consistently in CNT/F with larger lengths and diameters. CONCLUSION: Physicochemical characteristics, notably nominal CNT/F dimension and agglomerate size, predicted histopathologic changes and enabled grouping of materials by their toxicity profiles. Particles of greater nominal tube length were generally associated with increased severity of histopathology outcomes. Larger particle lengths and agglomerates were associated with more severe bronchi/bronchiolar outcomes. Spherical agglomerated particles of smaller nominal tube dimension were linked to granulomatous inflammation while a mixture of smaller and larger dimensional CNT/F resulted in more severe alveolar injury.

Physicochemical characterization and genotoxicity of the broad class of carbon nanotubes and nanofibers used or produced in U.S. facilities.

BACKGROUND: Carbon nanotubes and nanofibers (CNT/F) have known toxicity but simultaneous comparative studies of the broad material class, especially those with a larger diameter, with computational analyses linking toxicity to their fundamental material characteristics was lacking. It was unclear if all CNT/F confer similar toxicity, in particular, genotoxicity. Nine CNT/F (MW #1-7 and CNF #1-2), commonly found in exposure assessment studies of U.S. facilities, were evaluated with reported diameters ranging from 6 to 150 nm. All materials were extensively characterized to include distributions of physical dimensions and prevalence of bundled agglomerates. Human bronchial epithelial cells were exposed to the nine CNT/F (0-24 µg/ml) to determine cell viability, inflammation, cellular oxidative stress, micronuclei formation, and DNA double-strand breakage. Computational modeling was used to understand various permutations of physicochemical characteristics and toxicity outcomes. RESULTS: Analyses of the CNT/F physicochemical characteristics illustrate that using detailed distributions of physical dimensions provided a more consistent grouping of CNT/F compared to using particle dimension means alone. In fact, analysis of binning of nominal tube physical dimensions alone produced a similar grouping as all characterization parameters together. All materials induced epithelial cell toxicity and micronuclei formation within the dose range tested. Cellular oxidative stress, DNA double strand breaks, and micronuclei formation consistently clustered together and with larger physical CNT/F dimensions and agglomerate characteristics but were distinct from inflammatory protein changes. Larger nominal tube diameters, greater lengths, and bundled agglomerate characteristics were associated with greater severity of effect. The portion of tubes with greater nominal length and larger diameters within a sample was not the majority in number, meaning a smaller percentage of tubes with these characteristics was sufficient to increase toxicity. Many of the traditional physicochemical characteristics including surface area, density, impurities, and dustiness did not cluster with the toxicity outcomes. CONCLUSION: Distributions of physical dimensions provided more consistent grouping of CNT/F with respect to toxicity outcomes compared to means only. All CNT/F induced some level of genotoxicity in human epithelial cells. The severity of toxicity was dependent on the sample containing a proportion of tubes with greater nominal lengths and diameters.

In Vivo Toxicity Assessment of Occupational Components of the Carbon Nanotube Life Cycle To Provide Context to Potential Health Effects.

Pulmonary toxicity studies on carbon nanotubes focus primarily on as-produced materials and rarely are guided by a life cycle perspective or integration with exposure assessment. Understanding toxicity beyond the as-produced, or pure native material, is critical, due to modifications needed to overcome barriers to commercialization of applications. In the first series of studies, the toxicity of as-produced carbon nanotubes and their polymer-coated counterparts was evaluated in reference to exposure assessment, material characterization, and stability of the polymer coating in biological fluids. The second series of studies examined the toxicity of aerosols generated from sanding polymer-coated carbon-nanotube-embedded or neat composites. Postproduction modification by polymer coating did not enhance pulmonary injury, inflammation, and pathology or in vitro genotoxicity of as-produced carbon nanotubes, and for a particular coating, toxicity was significantly attenuated. The aerosols generated from sanding composites embedded with polymer-coated carbon nanotubes contained no evidence of free nanotubes. The percent weight incorporation of polymer-coated carbon nanotubes, 0.15% or 3% by mass, and composite matrix utilized altered the particle size distribution and, in certain circumstances, influenced acute in vivo toxicity. Our study provides perspective that, while the number of workers and consumers increases along the life cycle, toxicity and/or potential for exposure to the as-produced material may greatly diminish.

Elevated Concentrations of Lead in Particulate Matter on the Neighborhood-Scale in Delhi, India As Determined by Single Particle Analysis.

High mass concentrations of atmospheric lead particles are frequently observed in the Delhi, India metropolitan area, although the sources of lead particles are poorly understood. In this study, particles sampled across Delhi (August - December 2008) were analyzed by computer-controlled scanning electron microscopy with energy dispersive X-ray spectroscopy (CCSEM-EDX) to improve our understanding of the spatial and physicochemical variability of lead-rich particles (>90% lead). The mean mass concentration of lead-rich particles smaller than 10 µm (PM10) was 0.7 µg/m(3) (1.5 µg/m(3) std. dev.) with high variability (range: 0-6.2 µg/m(3)). Four samples (16% of 25 samples) with PM10 lead-rich particle concentrations >1.4 µg/m(3) were defined as lead events and studied further. The temporal characteristics, heterogeneous spatial distribution, and wind patterns of events, excluded regional monsoon conditions or common anthropogenic sources from being the major causes of the lead events. Individual particle composition, size, and morphology analysis indicate informal recycling operations of used lead-acid batteries as the likely source of the lead events. This source is not typically included in emission inventories, and the observed isolated hotspots with high lead concentrations could represent an elevated exposure risk in certain neighborhoods of Delhi.

Heterogeneity of coarse particles in an urban area.

The variation in composition and concentration of coarse particles in Rochester, a medium-sized city in western New York, was studied using UNC passive samplers and computer-controlled scanning electron microscopy (CCSEM). The samplers were deployed in a 5 × 5 grid (2 km × 2 km per grid cell) for 2-3 week periods in two seasons (September 2008 and May 2009) at 25 different sites across Rochester. CCSEM analysis yielded size and elemental composition for individual particles and analyzed more than 800 coarse particles per sample. Based on the composition as reflected in the fluoresced X-ray spectrum, the particles were grouped into classes with similar chemical compositions using an adaptive resonance theory (ART) network. The mass fractions of particles in the identified classes were then used to assess the homogeneity of composition and concentration across the measurement domain. These results illustrate how particle sampling using the UNC passive sampler coupled with CCSEM/ART can be used to determine the concentration and source of the coarse particulate matter at multiple sites. The particle compositions were dominated by elements suggesting that the major particle sources are road dust and biological particles. Considerable heterogeneity in both composition and concentration were observed between adjacent sites as indicated by cofficient of divergence analyses.

Measurements of OC and EC in coarse particulate matter in the southeastern United States.

The organic carbon (OC) and elemental carbon (EC) content of filter-based, 24-hr integrated particulate matter with aerodynamic diameters between 2.5 and 10 microm (PM10-2.5) was measured at two urban and two rural locations in the southeastern United States. On average, total carbon (OC + EC) comprised approximately 30% of PM10-2.5 mass at these four sites. Carbonate carbon was measured on a subset of samples from three sites and was found to be undetectable at a rural site in central Alabama, less than 2% of PM10-2.5 at an urban site in Georgia, and less than 10% of PM10-2.5 at an urban-industrial site in Alabama. Manual scanning electron microscopy (SEM) and computer-controlled SEM (CCSEM) along with energy dispersive X-ray spectroscopy (EDS) were used to identify individual carbonaceous particles in a selected subset of samples collected at one rural site and one urban-industrial site in Alabama. CCSEM results showed that biological material (e.g., fungal spores, pollen, and vegetative detritus) accounted for 60-70% of the carbonaceous mass in PM10-2.5 samples with concentrations in the range of 2-16 microg/m3. Samples with higher PM10-2.5 concentrations (25-42 microg/m3) at the urban-industrial site were found by manual SEM to have significant amounts of unidentified carbonaceous material, likely originating from local industrial activities. Both filter-based OC and EC concentrations and SEM-identified biological material tended to have higher concentrations during warmer months. Upper limits for organic mass (OM) to OC ratios (OM/OC) are estimated for PM10-2.5 samples at 2.1 for urban sites and 2.6-2.7 for rural sites.