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Inhalation Exposure to Carbon Nanotubes (CNT) and Carbon Nanofibers (CNF): Methodology and Dosimetry

Oberdörster, Günter, Castranova, Vincent, Asgharian, Bahman, Sayre, Phil·2015·Journal of Toxicology and Environmental Health, Part B
DOI 10.1080/10937404.2015.1051611PMC PMC4706753
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FIGURE 1 a. MWCNT aerosols at a research facility. Aerosols collected at a research facility during manufacture of MWCNT revealing a multiplicity of structures. (Han et al. 2008). Shown are samples collected in the facility after opening the furnace, demonstrating many non-fibrous structures in addition to some single fibers but mostly agglomerates of different sizes. Initially high concentrations could be significantly reduced following installation of proper enclosure and ventilation systems. b. MWCNT aerosols collected at a research facility. Aerosols collected at a research laboratory generating MWCNT by chemical vapor deposition (A: Tsai et al. 2009) and several commercial and research facilities manufacturing MWCNT (B, C, D: Methner et al. 2010). In the research facility, mostly agglomerates were seen and only some single filaments with numerous attached clusters of nanoparticles were found that consisted of carbon and included Fe from the catalyst (A). The aerosol in a commercial facility was described as “various forms like agglomerates, clusters, bundles, or nests rather than individual fibers or spherical particles” (B, C, D). These were collected under different working tasks (e.g., sawing, dumping). An example of the appearance of the few individual fibers and spherical particles is shown in (C). Total carbon concentrations measured on filters according to NIOSH NMAM Method 5040 (Issue 2, 1998) were equivalent to 1.8 mg/m3. In general, samples from both facilities indicate that long and individual fibers were hardly seen, in contrast to clusters/tangles of different sizes. The MWCNT aerosol generated experimentally for use in animal inhalation studies seems to be much better dispersed.
FIGURE 1 a. MWCNT aerosols at a research facility. Aerosols collected at a research facility during manufacture of MWCNT revealing a multiplicity of structures. (Han et al. 2008). Shown are samples collected in the facility after opening the furnace, demonstrating many non-fibrous structures in addition to some single fibers but mostly agglomerates of different sizes. Initially high concentrations could be significantly reduced following installation of proper enclosure and ventilation systems. b. MWCNT aerosols collected at a research facility. Aerosols collected at a research laboratory generating MWCNT by chemical vapor deposition (A: Tsai et al. 2009) and several commercial and research facilities manufacturing MWCNT (B, C, D: Methner et al. 2010). In the research facility, mostly agglomerates were seen and only some single filaments with numerous attached clusters of nanoparticles were found that consisted of carbon and included Fe from the catalyst (A). The aerosol in a commercial facility was described as “various forms like agglomerates, clusters, bundles, or nests rather than individual fibers or spherical particles” (B, C, D). These were collected under different working tasks (e.g., sawing, dumping). An example of the appearance of the few individual fibers and spherical particles is shown in (C). Total carbon concentrations measured on filters according to NIOSH NMAM Method 5040 (Issue 2, 1998) were equivalent to 1.8 mg/m3. In general, samples from both facilities indicate that long and individual fibers were hardly seen, in contrast to clusters/tangles of different sizes. The MWCNT aerosol generated experimentally for use in animal inhalation studies seems to be much better dispersed.CC BY-NC · PubMed Central
FIGURE 2 CNF aerosols collected on impactor stages during thermal treatment Transmission electron microscopy images of airborne CNF size selected by impactors located in a thermal treatment processing area for CNF. Impactor Stage A: 2.5–10 µm; Impactor Stage B: 1.0–2.5 µm; Impactor Stage C: 0.5–1.0 µm; Impactor Stage D: 0.25–0.5. Lacey carbon-coated Ni and SiO-coated Ni grids were used in impactors. (From: Birch et al. 2011).
FIGURE 2 CNF aerosols collected on impactor stages during thermal treatment Transmission electron microscopy images of airborne CNF size selected by impactors located in a thermal treatment processing area for CNF. Impactor Stage A: 2.5–10 µm; Impactor Stage B: 1.0–2.5 µm; Impactor Stage C: 0.5–1.0 µm; Impactor Stage D: 0.25–0.5. Lacey carbon-coated Ni and SiO-coated Ni grids were used in impactors. (From: Birch et al. 2011).CC BY-NC · PubMed Central
FIGURE 3 Dosimetric Extrapolation of Inhaled Particles from Rats to Humans Concept of dosimetric extrapolation of effects of inhaled materials in experimental animals to humans. A Human Equivalent Concentration (HEC) is derived based on study in rats (scenario 2 described in text) by using the MPPD model for rats to estimate the inhaled (delivered) and daily deposited dose in rats resulting from the specific experimental exposure concentrations. Knowing the material rat-specific clearance rates or retention halftime (T1/2) in the lung, the retained dose that has accumulated over the duration in the lung can be calculated, but may also be determined by analytical methods at the end of exposure. Using then human-specific clearance rates—if available—and breathing scenarios as inputs to the MPPD model for humans the HEC can be estimated for either the MMAD and GSD of the experimental aerosol, or for an aerosol size distribution that is known to be present for the human exposure scenario. The HEC is equivalent to the rat exposure in terms of resulting in the same deposited dose—or retained dose—in both species. Through using the MPPD model human and rat inhalability and respirability differences are taken into consideration. Deposited and retained dose can be expressed either per unit of epithelial surface in the respiratory tract—tracheobronchial or alveolar—, or per unit mass of the lung. In addition, the material based dosemetric can be mass (as indicated in the scheme), or surface area or number of CNT/CNF, if respective information is available. [From: (Oberdörster 1989)]
FIGURE 3 Dosimetric Extrapolation of Inhaled Particles from Rats to Humans Concept of dosimetric extrapolation of effects of inhaled materials in experimental animals to humans. A Human Equivalent Concentration (HEC) is derived based on study in rats (scenario 2 described in text) by using the MPPD model for rats to estimate the inhaled (delivered) and daily deposited dose in rats resulting from the specific experimental exposure concentrations. Knowing the material rat-specific clearance rates or retention halftime (T1/2) in the lung, the retained dose that has accumulated over the duration in the lung can be calculated, but may also be determined by analytical methods at the end of exposure. Using then human-specific clearance rates—if available—and breathing scenarios as inputs to the MPPD model for humans the HEC can be estimated for either the MMAD and GSD of the experimental aerosol, or for an aerosol size distribution that is known to be present for the human exposure scenario. The HEC is equivalent to the rat exposure in terms of resulting in the same deposited dose—or retained dose—in both species. Through using the MPPD model human and rat inhalability and respirability differences are taken into consideration. Deposited and retained dose can be expressed either per unit of epithelial surface in the respiratory tract—tracheobronchial or alveolar—, or per unit mass of the lung. In addition, the material based dosemetric can be mass (as indicated in the scheme), or surface area or number of CNT/CNF, if respective information is available. [From: (Oberdörster 1989)]CC BY-NC · PubMed Central
FIGURE 4 Exposure-Response and Dose-Response Exposure-Response and Dose-Response relationships of five 3-month inhalation studies in rats with MWCNT, CNF and CB. The carbon black (CB) study is added as a well studied low toxicity particle for comparison. Percent increase in lung lavage neutrophils (PMN) and retained lung dose were not reported for all of the five subchronic rat inhalation studies. Thus, to approximate retained lung burden of the MWCNT study by Ma-Hock et al. and for the CNF study by DeLorme et al., deposited and retained doses were calculated with the MPPD model, Version 2.90.3, using MMAD and GSD data and the packing density of the materials provided by the authors and a normal rat-specific pulmonary retention halftime (T1/2) of 70 days as model inputs. This T1/2 is appropriate for the rats exposed to the lower concentrations; for the high concentrations and resulting high lung burdens, a prolongation of T1/2 has to be expected (for example, as determined by Pauluhn et al., 2010, Table 3). Therefore, the lung burdens calculated for the high dose groups of these two studies are likely underestimated. The x-axes have been truncated to better separate the individual MWCNT/CNF data, although as a consequence the high exposure/date points could not be displayed. A: Lung weight and lung lavage neutrophil exposure-responses. B: Lung weight dose-responses based on retained lung burden expressed as mass, surface area and volume of the retained material. - MWCNT (Pauluhn, 2010); - MWCNT (Ma-Hock et al., 2009a); - CNF (DeLorme et al., 2012); - CB (Elder et al., 2005); - MWCNT (Kasai et al., 2015).
FIGURE 4 Exposure-Response and Dose-Response Exposure-Response and Dose-Response relationships of five 3-month inhalation studies in rats with MWCNT, CNF and CB. The carbon black (CB) study is added as a well studied low toxicity particle for comparison. Percent increase in lung lavage neutrophils (PMN) and retained lung dose were not reported for all of the five subchronic rat inhalation studies. Thus, to approximate retained lung burden of the MWCNT study by Ma-Hock et al. and for the CNF study by DeLorme et al., deposited and retained doses were calculated with the MPPD model, Version 2.90.3, using MMAD and GSD data and the packing density of the materials provided by the authors and a normal rat-specific pulmonary retention halftime (T1/2) of 70 days as model inputs. This T1/2 is appropriate for the rats exposed to the lower concentrations; for the high concentrations and resulting high lung burdens, a prolongation of T1/2 has to be expected (for example, as determined by Pauluhn et al., 2010, Table 3). Therefore, the lung burdens calculated for the high dose groups of these two studies are likely underestimated. The x-axes have been truncated to better separate the individual MWCNT/CNF data, although as a consequence the high exposure/date points could not be displayed. A: Lung weight and lung lavage neutrophil exposure-responses. B: Lung weight dose-responses based on retained lung burden expressed as mass, surface area and volume of the retained material. - MWCNT (Pauluhn, 2010); - MWCNT (Ma-Hock et al., 2009a); - CNF (DeLorme et al., 2012); - CB (Elder et al., 2005); - MWCNT (Kasai et al., 2015).CC BY-NC · PubMed Central
FIGURE 5 MWCNT aerosol size distribution in rodent inhalation study A: Subchronic inhalation of different types of cigarettes and air-exposed controls in nose-only tubes. B: Chronic inhalation of different types of cigarette smoke and diesel engine exhaust and air-exposed controls in nose-only tubes. Note: Effect of restraint stress on bodyweight in sham (clean air) exposed controls, highlighted in red.
FIGURE 5 MWCNT aerosol size distribution in rodent inhalation study A: Subchronic inhalation of different types of cigarettes and air-exposed controls in nose-only tubes. B: Chronic inhalation of different types of cigarette smoke and diesel engine exhaust and air-exposed controls in nose-only tubes. Note: Effect of restraint stress on bodyweight in sham (clean air) exposed controls, highlighted in red.CC BY-NC · PubMed Central
FIGURE 6 MWCNT aerosols collected in rodent inhalation study Aerosolized MWCNT collected on filter (A, B) and carbon-coated 200 mesh N: grids (C, D). The MWCNT were aerosolized using an acoustic aerosol generation system (McKinney, Chen, and Frazer 2009) for dry powder generation. Large variations of the airborne structures were observed, ranging from a few single tubes to differently-sized structures of agglomerated CNT to larger tangles resulting in an airborne size distribution as shown in Figure 5. The aerodynamic, settling and diffusional behavior cannot be inferred from these pictures; the large tangle in section D may not be inhalable by rodents and is likely not respirable, but can be estimated from their aerodynamic size distribution. Such experimentally generated MWCNT aerosol for rodent exposures needs to be compared to atmospheres found at workplaces. (See Figures 1, 2).
FIGURE 6 MWCNT aerosols collected in rodent inhalation study Aerosolized MWCNT collected on filter (A, B) and carbon-coated 200 mesh N: grids (C, D). The MWCNT were aerosolized using an acoustic aerosol generation system (McKinney, Chen, and Frazer 2009) for dry powder generation. Large variations of the airborne structures were observed, ranging from a few single tubes to differently-sized structures of agglomerated CNT to larger tangles resulting in an airborne size distribution as shown in Figure 5. The aerodynamic, settling and diffusional behavior cannot be inferred from these pictures; the large tangle in section D may not be inhalable by rodents and is likely not respirable, but can be estimated from their aerodynamic size distribution. Such experimentally generated MWCNT aerosol for rodent exposures needs to be compared to atmospheres found at workplaces. (See Figures 1, 2).CC BY-NC · PubMed Central
FIGURE 7 MWCNT aerosols collected in rodent inhalation study Aerosolized MWCNT collected on filter (A, B) and carbon-coated 200 mesh N: grids (C, D). The MWCNT were aerosolized using an acoustic aerosol generation system (McKinney et al., 2009) for dry powder generation. Large variations of the airborne structures were observed, ranging from a few single tubes to differently-sized structures of agglomerated CNT to larger tangles resulting in an airborne size distribution as shown in Figure 8. The aerodynamic, settling and diffusional behavior cannot be inferred from these pictures; the large tangle in section D may not be inhalable by rodents and is likely not respirable, but the aerodynamic behavior can be estimated from their aerodynamic size distribution. Such experimentally generated MWCNT aerosol for rodent exposures needs to be compared to atmospheres found at workplaces. (See Figures 1, 2).
FIGURE 7 MWCNT aerosols collected in rodent inhalation study Aerosolized MWCNT collected on filter (A, B) and carbon-coated 200 mesh N: grids (C, D). The MWCNT were aerosolized using an acoustic aerosol generation system (McKinney et al., 2009) for dry powder generation. Large variations of the airborne structures were observed, ranging from a few single tubes to differently-sized structures of agglomerated CNT to larger tangles resulting in an airborne size distribution as shown in Figure 8. The aerodynamic, settling and diffusional behavior cannot be inferred from these pictures; the large tangle in section D may not be inhalable by rodents and is likely not respirable, but the aerodynamic behavior can be estimated from their aerodynamic size distribution. Such experimentally generated MWCNT aerosol for rodent exposures needs to be compared to atmospheres found at workplaces. (See Figures 1, 2).CC BY-NC · PubMed Central
FIGURE 8 Novel method using hybrid markers to quantify MWCNT in tissues. a. Lung exposed to MWCNT. b. Transfer of lung tissue into C99 at room temperature. c. Lung digestion in C99. d. Lung solution centrifugation at 12,000 rpm for 10 minutes. e. Supernatant removal. f. Novel method using hybrid markers. Marker adsorbtion onto MWCNT for 15 min. g. Extraction of adsorbed B(ghi)P on MWCNTs to acetonitrile. h. Filtration of B(Ghi)P in the MWCNT. i. HPLC analysis. Limit of quantitation is 0.2 µg in MWCNT in rat lung.
FIGURE 8 Novel method using hybrid markers to quantify MWCNT in tissues. a. Lung exposed to MWCNT. b. Transfer of lung tissue into C99 at room temperature. c. Lung digestion in C99. d. Lung solution centrifugation at 12,000 rpm for 10 minutes. e. Supernatant removal. f. Novel method using hybrid markers. Marker adsorbtion onto MWCNT for 15 min. g. Extraction of adsorbed B(ghi)P on MWCNTs to acetonitrile. h. Filtration of B(Ghi)P in the MWCNT. i. HPLC analysis. Limit of quantitation is 0.2 µg in MWCNT in rat lung.CC BY-NC · PubMed Central
FIGURE 9 MWCNT aerosol size distribution in rodent inhalation study MWCNT aerosol size distribution as determined by a nano-MOUDI through weighing of deposits on the individual stages. The aerosol was generated with an acoustic aerosol generation system (Fig. 6) and collected with a 13-stage nano-MOUDI (NanoMoudi-II™ Model 125A, MSP Corp., Shoreview, MN) for 10 minutes at a flow rate of 10 l/min. Lower stages (10–13) were removed because deposits could not be reliably weighed; they contained less than 2% of the total mass. Larger structures with aerodynamic sizes >5 µm contribute much of the total mass concentration of the aerosol, but not to the deposition in the lower respiratory tract of rodents. Thus, describing a CNT/CNF aerosol exposure only by its total airborne mass concentration can be highly misleading when used as comparison to a human exposure concentration. Concepts of dosimetric rodent to human extrapolation discussed in this document have to be taken into consideration. Placing a cyclone separator in line before the aerosol enters the inhalation chamber may eliminate the non-inhalable tangles. On the other hand, if such larger structures are still inhalable by humans and if they are present at the workplace, it could be considered to use intratracheal inhalation (Oberdörster et al., 1997) in rats to circumvent the upper respiratory tract. However, intratracheal inhalation requires anesthesia of rats and is not recommended for day-to- day repeat exposures.
FIGURE 9 MWCNT aerosol size distribution in rodent inhalation study MWCNT aerosol size distribution as determined by a nano-MOUDI through weighing of deposits on the individual stages. The aerosol was generated with an acoustic aerosol generation system (Fig. 6) and collected with a 13-stage nano-MOUDI (NanoMoudi-II™ Model 125A, MSP Corp., Shoreview, MN) for 10 minutes at a flow rate of 10 l/min. Lower stages (10–13) were removed because deposits could not be reliably weighed; they contained less than 2% of the total mass. Larger structures with aerodynamic sizes >5 µm contribute much of the total mass concentration of the aerosol, but not to the deposition in the lower respiratory tract of rodents. Thus, describing a CNT/CNF aerosol exposure only by its total airborne mass concentration can be highly misleading when used as comparison to a human exposure concentration. Concepts of dosimetric rodent to human extrapolation discussed in this document have to be taken into consideration. Placing a cyclone separator in line before the aerosol enters the inhalation chamber may eliminate the non-inhalable tangles. On the other hand, if such larger structures are still inhalable by humans and if they are present at the workplace, it could be considered to use intratracheal inhalation (Oberdörster et al., 1997) in rats to circumvent the upper respiratory tract. However, intratracheal inhalation requires anesthesia of rats and is not recommended for day-to- day repeat exposures.CC BY-NC · PubMed Central
FIGURE A-1 CNT/CNF inhalability for humans and rats CNT/CNF inhalability for humans via oral and nasal, and rats nasal breathings. Equivalent impaction diameter in the figure includes shape and orientation effects. dei is found from Equation (1) for cylindrical shapes. Additional size measurements are needed to develop more realistic models of dei for CNT/CNF which, while being elongated, generally are not cylindrical.
FIGURE A-1 CNT/CNF inhalability for humans and rats CNT/CNF inhalability for humans via oral and nasal, and rats nasal breathings. Equivalent impaction diameter in the figure includes shape and orientation effects. dei is found from Equation (1) for cylindrical shapes. Additional size measurements are needed to develop more realistic models of dei for CNT/CNF which, while being elongated, generally are not cylindrical.CC BY-NC · PubMed Central
FIGURE B-1 CNT/CNF thoracic fractions for humans and rats CNT/CNF Thoracic fraction for different sizes for humans and rats at minute ventilations of 15 LPM and 7.4 mL/s, respectively.
FIGURE B-1 CNT/CNF thoracic fractions for humans and rats CNT/CNF Thoracic fraction for different sizes for humans and rats at minute ventilations of 15 LPM and 7.4 mL/s, respectively.CC BY-NC · PubMed Central
FIGURE B-2 Human to rat ratios for equal thoracic fractions Ratio of human equivalent concentration to rat exposure concentration based on equal thoracic fractions for minute ventilations of 15 LPM and 7.4 mL/s in humans and rats respectively.
FIGURE B-2 Human to rat ratios for equal thoracic fractions Ratio of human equivalent concentration to rat exposure concentration based on equal thoracic fractions for minute ventilations of 15 LPM and 7.4 mL/s in humans and rats respectively.CC BY-NC · PubMed Central
FIGURE B-3 CNT/CNF respiratory fractions for humans at different aspect ratios CNT/CNF deposition fraction in the human lung via nasal breathing for different aspect ratios. A minute ventilation of 7.5 LMP is used in the calculations.
FIGURE B-3 CNT/CNF respiratory fractions for humans at different aspect ratios CNT/CNF deposition fraction in the human lung via nasal breathing for different aspect ratios. A minute ventilation of 7.5 LMP is used in the calculations.CC BY-NC · PubMed Central
FIGURE B-4 HEC/CR ratios for CNT/CNF (100 nm—2 µm diameter) at aspect ratio of 10 Ratio of HEC/CR for different diameter CNT/CNF and aspect ratio of 10. Minute ventilations for humans and rats were 7.5 LPM and 0.241 LPM respectively.
FIGURE B-4 HEC/CR ratios for CNT/CNF (100 nm—2 µm diameter) at aspect ratio of 10 Ratio of HEC/CR for different diameter CNT/CNF and aspect ratio of 10. Minute ventilations for humans and rats were 7.5 LPM and 0.241 LPM respectively.CC BY-NC · PubMed Central

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Metadata

Authors
Oberdörster, G., Castranova, V., Asgharian, B., & Sayre, P.
Venue
Journal of Toxicology and Environmental Health, Part B
Publisher
Informa UK Limited
Published
2015-05-19
Volume
18
Issue
3-4
Pages
121-212
ISSN
  • 1093-7404
  • 1521-6950
DOI
10.1080/10937404.2015.1051611
PMC
PMC4706753
Cited by
150
References
6
Language
en

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CoChat. (2026). Citation check: Inhalation Exposure to Carbon Nanotubes (CNT) and Carbon Nanofibers (CNF): Methodology and Dosimetry. https://app.cochat.ai/papers/10.1080/10937404.2015.1051611
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Oberdörster, G., Castranova, V., Asgharian, B., & Sayre, P. (2015). Inhalation Exposure to Carbon Nanotubes (CNT) and Carbon Nanofibers (CNF): Methodology and Dosimetry. Journal of Toxicology and Environmental Health, Part B. https://doi.org/10.1080/10937404.2015.1051611