length, respectively) [Muller et al. 2009; Liang et al.2010] did not produce mesothelioma. These findingsare consistent with those reported by Yamashitaet al. [2010] <strong>and</strong> Nagai et al. [2011] who foundthat MWCNT injected in<strong>to</strong> the peri<strong>to</strong>neal cavity ofmice or rats generated inflammation/genetic damage<strong>and</strong> mesothelioma that were related <strong>to</strong> the dimensionof the CNT. Results from these peri<strong>to</strong>nealassay studies indicate that CNT of specific dimensions<strong>and</strong> durability can cause inflammation, fibrosis,<strong>and</strong> mesothelial tumors in mice <strong>and</strong> in rats;however, additional experimental animal researchis needed <strong>to</strong>: (1) provide quantitative data on thebiopersistence of different types of CNT in the lung<strong>and</strong>, (2) address the key question as <strong>to</strong> the precisedimensions (<strong>and</strong> possibly other physical-chemicalcharacteristics) of CNT that pose a potential pathogenicrisk for cancer including mesothelioma.As synthesized, raw (unpurified) CNT, contain asmuch as 30% catalytic metals. Catalytic metals,such as iron-rich SWCNT, can generate hydroxylradicals in the presence of hydrogen peroxide <strong>and</strong>organic (lipid) peroxides [Kagan et al. 2006], <strong>and</strong>when human epidermal keratinocytes cells are exposed<strong>to</strong> unpurified SWCNT (in vitro cellular studies),oxidant injury occurs [Shvedova et al. 2003].These catalytic metals can be removed from rawCNT by acid treatment or by high temperature <strong>to</strong>yield purified CNT with low metal content. Removalof catalytic metals abolishes the ability ofSWCNT or MWCNT <strong>to</strong> generate hydroxyl radicals.However, in labora<strong>to</strong>ry animal studies the pulmonarybioactivity of SWCNT does not appear <strong>to</strong>be affected by the presence or absence of catalyticmetals. Lam et al. [2004] compared the pulmonaryresponse of mice <strong>to</strong> intratracheal instillation of raw(containing 25% metal catalyst) with purified (~2%iron) SWCNT <strong>and</strong> found that the granuloma<strong>to</strong>usreaction was not dependent on metal contamination.Likewise, the acute inflamma<strong>to</strong>ry reaction ofmice after aspiration of raw (30% iron) versus purified(< 1% iron) SWCNT was not affected by metalcontent [Shvedova et al. 2005, 2008].Pulmonary exposure <strong>to</strong> CNT have shown systemicresponses including an increase in inflamma<strong>to</strong>rymedia<strong>to</strong>rs in the blood, as well as oxidant stress inaortic tissue <strong>and</strong> increase plaque formation in anatherosclerotic mouse model [Li et al. 2007; Erdelyet al. 2009]. Pulmonary exposure <strong>to</strong> MWCNTalso depresses the ability of coronary arterioles <strong>to</strong>respond <strong>to</strong> dila<strong>to</strong>rs [Staple<strong>to</strong>n et al. 2011]. Thesecardiovascular effects may be due <strong>to</strong> neurogenicsignals from sensory irritant recep<strong>to</strong>rs in the lung.Mechanisms, such as inflamma<strong>to</strong>ry signals or neurogenicpathways causing these systemic responses,are under investigation.Results from in vitro cellular studies have shownthat SWCNT can cause geno<strong>to</strong>xicity <strong>and</strong> abnormalchromosome number, because of interference withmi<strong>to</strong>sis (cell division), by disrupting the mi<strong>to</strong>ticspindles in dividing cells <strong>and</strong> inducing the formationof anaphase bridges among the nuclei [Sargentet al. 2009]. In vitro studies also indicate that exposure<strong>to</strong> CNF can cause geno<strong>to</strong>xicity (micronuclei)as a result of reactive oxygen species (ROS) production,which in turn reacts with DNA, <strong>and</strong> byinterfering physically with the DNA/chromosomes<strong>and</strong>/or mi<strong>to</strong>tic apparatus [Kisin et al. 2011]. Lowdose,long-term exposure of bronchial epithelialcells <strong>to</strong> MWCNT has been shown <strong>to</strong> induce celltransformation, <strong>and</strong> these transformed cells inducetumors after injection in<strong>to</strong> nude mice [Stueckle etal. 2011; Wang et al. 2011].Currently, there are no studies reported in the literatureon the adverse health effects in workers producingor using CNT or CNF. However, becausehumans can also develop lung inflammation <strong>and</strong>fibrosis in response <strong>to</strong> inhaled particles <strong>and</strong> fibers,it is reasonable <strong>to</strong> assume that at equivalent exposures(e.g., lung burden/alveolar epithelial cell surface)<strong>to</strong> CNT <strong>and</strong> CNF, workers may also be at riskof developing these adverse lung effects.Although data on workplace exposures <strong>to</strong> CNT <strong>and</strong>CNF are limited, aerosolization of CNT <strong>and</strong> CNFhas been shown <strong>to</strong> occur at a number of operationsduring research, production, <strong>and</strong> use of CNT <strong>and</strong>CNF, including such work tasks as transferring,weighing, blending, <strong>and</strong> mixing. Worker exposure<strong>to</strong> airborne CNT <strong>and</strong> CNF has frequently been34 NIOSH CIB 65 • <strong>Carbon</strong> <strong>Nanotubes</strong> <strong>and</strong> <strong>Nanofibers</strong>
observed <strong>to</strong> be task-specific <strong>and</strong> short-term in duration,with exposure concentrations (frequentlyreported as particle number or mass concentrations)found <strong>to</strong> exceed background exposure measurementswhen appropriate engineering controlsare not used <strong>to</strong> reduce exposures [Maynard et al.2004; Methner et al. 2007; Han et al. 2008a; Bello etal. 2009; Tsai et al. 2009; Bello et al. 2010; Evans et al.2010; Johnson et al. 2010; Lee et al. 2010; Cena <strong>and</strong>Peters 2011; Dahm et al. 2011]. Results from studiesalso suggest that the airborne concentration <strong>and</strong>the physical-chemical characteristics of particles(e.g., discrete versus agglomerated CNT) releasedwhile h<strong>and</strong>ling CNT may vary significantly withproduction batch <strong>and</strong> work process. Comprehensiveworkplace exposure evaluations are needed <strong>to</strong>characterize <strong>and</strong> quantify worker exposure <strong>to</strong> CNT<strong>and</strong> CNF at various job tasks <strong>and</strong> operations, <strong>and</strong><strong>to</strong> determine what control measures are the mosteffective in reducing worker exposures.The findings of adverse respira<strong>to</strong>ry effects (i.e., pulmonaryfibrosis, granuloma<strong>to</strong>us inflammation) <strong>and</strong>systemic responses in animals indicate the needfor protective measures <strong>to</strong> reduce the health risk <strong>to</strong>workers exposed <strong>to</strong> CNT <strong>and</strong> CNF. Availableevidence also indicates that the migration of MW-CNT in<strong>to</strong> the intrapleural space could potentiallyinitiate mesothelial injury <strong>and</strong> inflammation tha<strong>to</strong>ver time cause pleural pathology, including mesothelioma.Long-term inhalation studies are needed<strong>to</strong> determine whether CNT <strong>and</strong> CNF of specificdimension <strong>and</strong> chemistry can cause cancer inlabora<strong>to</strong>ry animals at doses equivalent <strong>to</strong> potentialworkplace exposures. In addition, the potential formigration of CNT through the lungs <strong>and</strong> for accumulationin the intrapleural space with time afterinhalation requires further investigation. Until resultsfrom animal research studies can fully explainthe mechanisms in which inhalation exposure <strong>to</strong>CNT <strong>and</strong> CNF cause adverse lung effects <strong>and</strong> possiblesystemic effects, all types of CNT <strong>and</strong> CNFshould be considered an occupational respira<strong>to</strong>ryhazard, <strong>and</strong> the following actions should be taken<strong>to</strong> minimize health concerns:1. Minimize workplace exposures.2. Establish an occupational health surveillanceprogram for workers exposed <strong>to</strong> CNT <strong>and</strong> CNF(Section 6, Appendix B).NIOSH CIB 65 • <strong>Carbon</strong> <strong>Nanotubes</strong> <strong>and</strong> <strong>Nanofibers</strong>35
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CURRENT INTELLIGENCE BULLETIN 65Occ
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Current Intelligence Bulletin 65Occ
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ForewordThe Occupational Safety and
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Executive SummaryOverviewCarbon nan
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- Page 17 and 18: Periodic Evaluations••Evaluatio
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- Page 25 and 26: AcknowledgementsThis Current Intell
- Page 27 and 28: 1 IntroductionMany nanomaterial-bas
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- Page 36 and 37: information on air contaminants. Sa
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- Page 45 and 46: 3.3 SWCNT and MWCNTIntraperitoneal
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- Page 97 and 98: 7 Research NeedsAdditional data and
- Page 99 and 100: ReferencesACGIH [1984]. Particle si
- Page 101 and 102: Bolton RE, Vincent HJ, Jones AD, Ad
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Piegorsch WW, Bailer AF [2005]. Qua
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AD, Baron PA [2003]. Exposure to ca
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Varga C, Szendi K [2010]. Carbon na
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ContentsA.1 Introduction ..........
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A.1 IntroductionThe increasing prod
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provide an informal check on the es
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these same dose groups; this effect
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Table A-1. Rodent study information
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the deposited (no clearance) and th
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The other BMDS models failed to con
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Figure A-2. Benchmark dose model (m
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Figure A-3 (continued). Benchmark d
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Table A-3. Benchmark dose estimates
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Table A-5. Benchmark dose estimates
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histopathology grade 2 or higher lu
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Table A-8. Working lifetime percent
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developing early-stage adverse lung
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Figure A-4. Dose-response relations
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cell surface area). However, the wo
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purified or unpurified (with differ
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Table A-9. Comparison of rat or hum
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A.6.1.3 Pulmonary Ventilation RateT
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used as the effect levels in evalua
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the DF estimate, although a larger
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or overloading, of particle clearan
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Table A-13. Human-equivalent retain
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A.7.1 Particle CharacteristicsBoth
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and density. The following MMAD and
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Table A-15. CNT lung dose normalize
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B.1 Key Terms Related toMedical Sur
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APPENDIX CNIOSH Method 5040
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filter. In the method evaluation, d
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Most of the studies on sampling art
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e analyzed to determine the onset o
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Delivering on the Nation’s promis