Final Report: Risk Assessment for Manufactured Nanoparticles Used in Consumer Products (RAMNUC)

EPA Grant Number: R834693
Title: Risk Assessment for Manufactured Nanoparticles Used in Consumer Products (RAMNUC)
Investigators: Zhang, Junfeng , Chung, Kian Fan , Di Giulio, Richard T. , Garfunkel, Eric , Georgopoulos, Panos G. , Isukapalli, Sastry S. , Kipen, Howard , Lee, Ki-Bum , Lioy, Paul J. , Mainelis, Gediminas , Porter, Alexandra , Ryan, Mary P. , Schwander, Stephan K. , Tetley, Teresa D
Institution: University of Medicine and Dentistry of New Jersey , Duke University , Imperial College
EPA Project Officer: Lasat, Mitch
Project Period: April 1, 2011 through June 30, 2014 (Extended to June 30, 2016)
Project Amount: $1,999,995
RFA: Environmental Behavior, Bioavailability and Effects of Manufactured Nanomaterials - Joint US – UK Research Program (2009) RFA Text |  Recipients Lists
Research Category: Nanotechnology , Safer Chemicals

Objective:

The overall objective of the RAMNUC project is to assess physicochemical and toxicological properties of manufactured nanoparticles at the point of exposure in comparison to those properties at the source (nanoparticles synthesized in the laboratory or materials acquired commercially). The differences between the source and the point of exposure may have significant consequences with respect to MNPs’ bioavailability, alterations of immunity, induction of oxidative stress, inflammation, disease processes, and other toxicity measures. Using selected consumer products that respectively incorporate zinc oxide, silver, and cerium dioxide nanoparticles, the RAMNUC project aims to understand how these nanoparticles transform as they enter and move through the atmosphere and when inhaled.

Summary/Accomplishments (Outputs/Outcomes):

Aerosolization of manufactured nanoparticles in consumer products: Exposures to airborne particles ranging from 14 nm to 20 µm due to the use of nanotechnology-based cosmetic powders were studied realistically by applying to a mannequin’s face while measuring the concentration and size distribution of inhaled aerosol particles. It was calculated that the highest inhaled particle mass was in the coarse aerosol fraction (2.5–10 µm), while particles <100 nm made a minimal contribution to the inhaled particle mass. For all powders, 85–93 % of aerosol deposition occurred in the head airways, while <10 % deposited in the alveolar and <5 % in the tracheobronchial regions. Electron microscopy data suggest that nanomaterials were present in the form of agglomerates across the entire investigated aerosol size range (14 nm–10 µm). Thus, the nanoparticle health effects should consider not only the alveolar region, but also other respiratory system regions where substantial nanomaterial deposition during the actual nanotechnology-based product use would occur. This particle size distribution would reduce the potential of the inhaled particles to enter the blood stream.

Characterization of silver, zinc and cerium nanoparticles in consumer products: The bioreactivity of nanoparticles in cells isolated from human lung tissue varied with both the type of particle (e.g., zinc oxide or silver) and the carrier solution. Through the characterization of how silver nanoparticles transform following incubation with cells, it was found that the lung lining fluid, particularly dipalmitoylphosphatidylcholine (DPPC), modified the kinetics of silver ion release from silver nanoparticles. Silver nanowires, inhaled into the lung and inside the human alveolar cells, were dissolved and subsequently transformed into the highly insoluble silver sulfide. When the lung cells were exposed to a commercially-available antifungal spray consisting of nano silver suspended in water, toxic effects were not observed; neither were they observed when the nano-silver and water were separated from each other and tested alone. However, when a commercially-available cleaning product, also containing nano silver, was tested, it exhibited significant toxic effects. These results suggest an increased inflammatory response due to the cleaning product solvent alone and a possible toxic effect of silver nanoparticles due to interaction with the solvent. The solvent-dependent effect demonstrates the importance of testing nanomaterials as they occur in commercial products themselves, as experienced by the consumer, as well as in isolation. A similar finding was reported for zinc-containing sprays, where the relative toxicity varied between products and depended on the type of spray involved. Overall, these studies suggest that inhalation of nanoparticle-containing sprays could have a harmful health effect and that the solvent used in the product can change the fate, behavior and toxicity of the nanoparticles in these products.

Effects of silver, zinc and cerium nanoparticles in vitro and in vivo: In studies of consumer spray products containing silver and zinc nanoparticles, the RAMNUC discovered that the bioreactivity of the product depended on the type of particle and the medium (i.e., carrier solution). Neither the silver particles, nor the medium of Mesosilver (a tested consumer product) was toxic to respiratory lung cells in vitro. In contrast, for Nanofix (a tested consumer product), which also contained silver particles, the silver particles were toxic alone, as was high concentration of the medium alone, resulting in a very toxic mixture. Therazinc (a tested consumer product) was also found to be very toxic, largely due to the medium, but also at high particle concentrations. Conversely, the nanoparticles in Dermazinc (a tested consumer product) were most toxic, whereas the medium was toxic only at high concentrations, making the whole product very toxic. These studies indicate that even if these sprays are used for other purposes (e.g., cleaning and skin conditioning), unintended inhalation could have adverse health effects.

In another set of experiments, RAMNUC examined the potential toxic effects of silver nanoparticles generated from a spark generator (mass concentration: 600-800 μg/mm3; mean diameter: 13-16 nm; total lung doses: 8 and 26-28 μg) inhaled by the nasal route in two rat strains. Results showed that cellular inflammation and increasing silver-positive macrophages were present in the lungs at day 7, and it was associated with significant levels of lung silver which indicate that lung toxicity is persistent for up to one week. In Brown-Norway rat strain, a pre-existing inflammatory condition is likely to lead to an increased amount of silver retained in the lungs resulting in parenchymal dysfunction. This was not observed in Sprague-Dawley strain. The pre-existing inflammatory state of the Brown-Norway rat is likely to underlie the increased amount of silver retained in the lungs with a reduced clearance rate that may underlie the increased inflammatory response, induction of surfactant protein D and phospholipids, and airway and parenchymal dysfunction observed in this rat strain but not in Sprague–Dawley strain. This would indicate that inhalation of AgNPs in people with pre-existing inflammation in the lungs such as asthma patients or those with chronic obstructive pulmonary disease would lead to a greater degree of inflammation with heightened consequences on lung function.

Interaction of pulmonary surfactant with zinc oxide (ZnO) nanoparticles: Alveolar lung lining liquid containing surfactant is the first extracellular barrier that inhaled nanoparticles will encounter in the respiratory units. The underlying alveolar epithelium is therefore likely to interact with surfactant-modified nanoparticles. Pulmonary surfactant can significantly alter the dissolution kinetics, aggregation state and surface chemistry of ZnO nanowires (ZnONW), with important consequences on how underlying epithelial cells internalized and processed these nanowires. Results showed that combinations of spatially resolved static and dynamic techniques are required to develop a holistic understanding of the parameters that govern ZnONW bioreactivity at the point of exposure and to accurately predict their risks on human health and the environment. In vitro adsorption of pulmonary surfactant lipids on ZnONWs has been demonstrated for the first time. The lipid corona delayed the kinetics of Zn2+ ion release from ZnONWs at acidic pH, by blocking direct contact between the nanowire surface and the aqueous environment. Pulmonary surfactant prevented the agglomeration of ZnONWs, possibly through contributions of both steric and charge stabilization. The role of pulmonary surfactant is central in our understanding interactions at the bio-nano interface of the alveoli, and their impact on subsequent epithelial–endothelial nanoparticle translocation.

Toxicological effects of cerium oxide nanoparticles in diesel fuel: One research focus of the RAMNUC consortium was to investigate the fate, behavior and toxicological effects of ceria nanoparticles as a diesel fuel additive. It was found that adding ceria reduced emission rates of pollutants including carbon monoxide, carbon dioxide, total particle mass, formaldehyde, acetaldehyde, acrolein and several polycyclic hydrocarbons. However, there was also an increase in emission rates for oxides of nitrogen and ultrafine particle counts. The addition of nano-ceria in the diesel fuel also affected several physicochemical properties of diesel exhaust particles (DEPs), e.g., reducing particle size, reducing carbon content, increasing cerium content, increasing organic carbon to elemental carbon ratio, and reducing oxidation potential measured as ascorbate depletion rate. These changes in physiochemical properties may be responsible for the changes in toxicity and bioreactivity of diesel exhaust particles (DEPs) observed in four experimental models. First, when cultured human lung cells were exposed to ceria-containing DEPs, the inflammatory response was reduced compared with those from untreated diesel, particularly at the higher concentrations of ceria, suggesting an anti-inflammatory effect. Second, a similar reduction in inflammatory response by DEPs from the nano-ceria treated fuel compared to the untreated fuel in mice instilled with DEPs was observed. Third, a significant, more than two-fold, decrease in mortality rate was observed when dechorionated zebrafish embryos were exposed to DEP from the ceria treated fuel compared to DEP from the untreated fuel. Lastly, studies of the effect of the ceria additive on immune responses in blood monocytes found that the effect was linked to changes in size and zeta potential of DEP induced by nano-ceria.

Modelling risks and biological responses: The sheer number of nanoparticle-containing products, application and uses, and the vast range of organisms that could be affected present significant challenges to the prediction of health risk. The task is further complicated by their complex transformation upon entering the environment. To respond to these challenges and to capture realistic exposure scenarios at a population level, RAMNUC has taken a modular modelling approach that includes use of a geographic information system and particle size data to identify possible exposure to nanoparticles in both indoor and outdoor environments.

To characterize ambient and indoor population exposures to silver nanoparticles, a novel tiered modeling system, Prioritization/Ranking of Toxic Exposures with GIS Extension (PRoTEGE), was developed and implemented for ambient and indoor environments, utilizing available data for nanoparticles production, usage, and property databases. The data were complemented with laboratory measurements of potential exposures from nano silver-containing consumer spray products generated by RAMNUC. Modeling of micro-environmental levels of nanomaterials employs Probabilistic Material Flow Analysis combined with product Life Cycle Analysis to account for releases during manufacturing, transport, usage, disposal, etc. Population distributions of intakes, consistent with published individual-based intake estimates, were used to calculate biologically relevant population distributions of uptakes and target tissue doses through human airway dosimetry modeling taking into account nanoparticle size distributions and age-relevant physiological parameters. Mathematical models of nanoparticle fate, behavior, and “in vitro” toxicity were developed to support analysis and prediction of in vivo effects of nanoparticles by predicting changes in cellular mechanisms. RAMNUC investigated the effects of particle properties on biological toxicity using the Agglomeration-diffusion-sedimentation-reaction model (ADSRM) and used a direct simulation Monte Carlo method to study the transformation of nanoparticles in biological media. Nanoparticle diffusion, gravitational settling, agglomeration, and dissolution were treated in a mechanistic manner. The model predictions for agglomeration and dissolution were compared with in vitro measurements for various types of MNPs, coating materials, and incubation media, and were found to be consistent with the measurements. Modeling of fate, transport, and effects of nanoparticles throughout the lung was used to predict particle transport across the air/biological fluid interface and the final expected dose for both coated and uncoated particles.

 

Conclusions:

The toxicity and bioreactivity of MNPs collected at the point of exposure or in the form of consumer products are different from those of MNPs as synthesized or in the original form before added into a consumer product. The difference is attributable to the changes in physical and chemical properties of MNPs when they are formulated into the consumer product, released into the atmosphere, and/or after enter the respiratory tract. RAMNUC has demonstrated the predictability of a source-to-dose-to-effect modular modeling system. Population exposures and inhaled doses can be mathematically modeled, along with toxicity data, to predict health risks associated with the use of MNPs incorporated in consumer products.

 

RAMNUC is a product of a collaborative effort funded through the US-UK joint program on nanotechnology research. This allowed researchers from two US institutions (Duke University and Rutgers University and two UK institutions (Imperial College London and Public Protection Agency of England) to work together and share knowledge across national borders in an effort to understand the complexities associated with MNP toxicity an population exposure. The synergetic expertise of the RAMNUC team and findings from this research have facilitated the successful competition of several grants and supplementary funds. These additional grant supports, in turn, have deepened and expanded the scope of the research initially proposed in the RAMNUC proposal, leading to a long list of publications listed below.

The output of RAMNUC reflects trans-Atlantic perspectives on risks associated with MNPs used in consumer products. For example, nano CeO2 based diesel additives have been used in the UK and elsewhere in the Europe, but not yet in the United States. Findings from RAMNUC suggest that the use of such an additive can reduce the emissions of certain air pollutants as well as the bioreactivity and toxicity of diesel exhaust particles. If the findings can be reproduced or confirmed in future studies using diesel engines used in vehicles, the use of nano-CeO2 fuel additive in the US diesel fleet should result in both climate and health benefits through reduced emissions of CO2 and diminished toxicity of diesel exhaust particles.


Journal Articles on this Report : 24 Displayed | Download in RIS Format

Other project views: All 41 publications 24 publications in selected types All 24 journal articles
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Journal Article Calderon L, Han T, McGilvery CM, Yang L, Subramaniam P, Lee K-B, Schwander S, Tetley TS, Georgopoulos P, Ryan M, Porter AE, Smith R, Chung KF, Lioy PJ, Zhang J, Mainelis G. Release of airborne particles and Ag and Zn compounds from nanotechnology-enabled consumer sprays: Implications for inhalation exposure. Atmospheric Environment. 2017;155:85-96. R834693 (Final)
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  • Journal Article Chen S, Theodorou IG, Goode AE, Gow A, Schwander S, Zhang JJ, Chung KF, Tetley TD, Shaffer MS, Ryan MP, Porter AE. High-resolution analytical electron microscopy reveals cell culture media-induced changes to the chemistry of silver nanowires. Environmental Science & Technology 2013;47(23):13813-13821. R834693 (2014)
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  • Journal Article Chen S, Goode AE, Sweeney S, Theodorou IG, Thorley AJ, Ruenraroengsak P, Chang Y, Gow A, Schwander S, Skepper J, Zhang JJ, Shaffer MS, Chung KF, Tetley TD, Ryan MP, Porter AE. Sulfidation of silver nanowires inside human alveolar epithelial cells: a potential detoxification mechanism. Nanoscale 2013;5(20):9839-9847. R834693 (2014)
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  • Journal Article Chen S, Goode AE, Skepper JN, Thorley AJ, Seiffert JM, Chung KF, Tetley TD, Shaffer MS, Ryan MP, Porter AE. Avoiding artefacts during electron microscopy of silver nanomaterials exposed to biological environments. Journal of Microscopy 2015;261(2):157-166. R834693 (Final)
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  • Journal Article Cole TB, Fisher JC, Burbacher TM, Costa LG, Furlong CE. Neurobehavioral assessment of mice following repeated postnatal exposure to chlorpyrifos-oxon. Neurotoxicology and Teratology 2012;34(3):311-322. R834693 (Final)
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  • Journal Article Leo BF, Chen S, Kyo Y, Herpoldt KL, Terrill NJ, Dunlop IE, McPhail DS, Shaffer MS, Schwander S, Gow A, Zhang J, Chung KF, Tetley TD, Porter AE, Ryan MP. The stability of silver nanoparticles in a model of pulmonary surfactant. Environmental Science & Technology 2013;47(19):11232-11240. R834693 (2014)
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  • Journal Article Lopez-Heras M, Theodorou IG, Leo BF, Ryan MP, Porter AE. Towards understanding the antibacterial activity of Ag nanoparticles: electron microscopy in the analysis of the materials-biology interface in the lung. Environmental Science: Nano 2015;2(4):312-326. R834693 (Final)
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  • Journal Article Nazarenko Y, Zhen H, Han T, Lioy PJ, Mainelis G. Potential for inhalation exposure to engineered nanoparticles from nanotechnology-based cosmetic powders. Environmental Health Perspectives 2012;120(6):885-892. R834693 (2012)
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  • Journal Article Nazarenko Y, Zhen H, Han T, Lioy PJ, Mainelis G. Nanomaterial inhalation exposure from nanotechnology-based cosmetic powders: a quantitative assessment. Journal of Nanoparticle Research 2012;14(11):1229 (14 pp.). R834693 (2014)
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  • Journal Article Nazarenko Y, Lioy PJ, Mainelis G. Quantitative assessment of inhalation exposure and deposited dose of aerosol from nanotechnology-based consumer sprays. Environmental Science: Nano 2014;1(2):161-171. R834693 (2013)
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  • Journal Article Royce SG, Mukherjee D, Cai T, Xu SS, Alexander JA, Mi Z, Calderon L, Mainelis G, Lee K, Lioy PJ, Tetley TD, Chung KF, Zhang J, Georgopoulos PG. Modeling population exposures to silver nanoparticles present in consumer products. Journal of Nanoparticle Research 2014;16(11):2724. R834693 (2013)
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  • Journal Article Sarkar S, Zhang L, Subramaniam P, Lee KB, Garfunkel E, Strickland PA, Mainelis G, Lioy PJ, Tetley TD, Chung KF, Zhang J, Ryan M, Porter A, Schwander S. Variability in bioreactivity linked to changes in size and zeta potential of diesel exhaust particles in human immune cells. PLoS ONE 2014;9(5):e97304 (12 pp.). R834693 (2014)
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  • Journal Article Seiffert J, Hussain F, Wiegman C, Li F, Bey L, Baker W, Porter A, Ryan MP, Chang Y, Gow A, Zhang J, Zhu J, Tetley TD, Chung KF. Pulmonary toxicity of instilled silver nanoparticles: influence of size, coating and rat strain. PLOS ONE 2015;10(3):e0119726 (17 pp.). R834693 (2013)
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  • Journal Article Seiffert J, Buckley A, Leo B, Martin NG, Zhu J, Dai R, Hussain F, Guo C, Warren J, Hodgson A, Gong J, Ryan MP, Zhang JJ, Porter A, Tetley TD, Gow A, Smith R, Chung KF. Pulmonary effects of inhalation of spark-generated silver nanoparticles in Brown-Norway and Sprague-Dawley rats. Respiratory Research 2016;17(1):85 (15 pp.). R834693 (2014)
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  • Journal Article Subramaniam P, Lee SJ, Shah S, Patel S, Starovoytov V, Lee K-B. Generation of a library of non-toxic quantum dots for cellular imaging and siRNA delivery. Advanced Materials 2012;24(29):4014-4019. R834693 (2012)
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  • Journal Article Theodorou IG, Ryan MP, Tetley TD, Porter AE. Inhalation of silver nanomaterials--seeing the risks. International Journal of Molecular Sciences 2014;15(12):23936-23974. R834693 (Final)
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  • Journal Article Theodorou IG, Botelho D, Schwander S, Zhang J, Chung KF, Tetley TD, Shaffer MS, Gow A, Ryan MP, Porter AE. Static and dynamic microscopy of the chemical stability and aggregation state of silver nanowires in components of murine pulmonary surfactant. Environmental Science & Technology 2015; 49(13):8048-8056. R834693 (Final)
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  • Journal Article Zhang J, Nazarenko Y, Zhang L, Calderon L, Lee KB, Garfunkel E, Schwander S, Tetley TD, Chung KF, Porter AE, Ryan M, Kipen H, Lioy PJ, Mainelis G. Impacts of a nanosized ceria additive on diesel engine emissions of particulate and gaseous pollutants. Environmental Science & Technology 2013;47(22):13077-13085. R834693 (2013)
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  • Journal Article Zhang J, Lee K-B, He L, Seiffert J, Subramaniam P, Yang L, Chen S, Maguire P, Mainelis G, Schwander S, Tetley T, Porter, A, Ryan M, Shaffer M, Hu S, Gong J, Chung KF. Effects of a nanoceria fuel additive on physicochemical properties of diesel exhaust particles. Environmental Science: Processes and Impacts 2016;18(10):1333-1342. R834693 (Final)
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  • Journal Article Mukherjee D, Leo BF, Royce SF, Porter AE, Ryan MP, Schwander S, Chung KF, Tetley TD, Zhang J, Georgopoulos PG. Modeling physiochemical interactions affecting in vitro cellular dosimetry of engineered nanomaterials: application to nanosilver. Journal of Nanoparticle Research 2014;16(10):2616 (29 pp.). R834693 (Final)
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  • Journal Article Mukherjee D, Royce SG, Sarkar S, Thorley A, Schwander S, Ryan MP, Porter AE, Chung KF, Tetley TD, Zhang J, Georgopoulos PG. Modeling in vitro cellular responses to silver nanoparticles. Journal of Toxicology 2014;2014:852890 (13 pp.). R834693 (Final)
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  • Journal Article Mukherjee D, Porter, A, Ryan M, Schwander S, Chung KF, Tetley T, Zhang J, Georgopoulos P. Modeling in vivo interactions of engineered nanoparticles in the pulmonary alveolar lining fluid. Nanomaterials 2015;5(3):1223-1249. R834693 (Final)
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  • Journal Article Theodorou IG, Ruenraroengsak P, Gow A, Schwander S, Zhang JJ, Chung KF, Tetley TD, Ryan MP, Porter AE. Effect of pulmonary surfactant on the dissolution, stability and uptake of zinc oxide nanowires by human respiratory epithelial cells. Nanotoxicology 2016;10(9):1351-1362. R834693 (Final)
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  • Journal Article Chung KF, Seiffert J, Chen S, Theodorou I, Goode A, Leo BF, McGilvery C, Hussain F, Wiegman C, Rossios C, Zhu J, Gong J, Tariq F, Yufit V, Monteith A, Hashimoto T, Skepper J, Ryan M, Zhang J, Tetley T, Porter A. Inactivation, clearance and functional effects of lung-instilled short and long silver nanowires in rat. ACS Nano 2017;11(3):2652-2664. R834693 (Final)
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  • Supplemental Keywords:

    silver, zinc oxide, cerium dioxide, diesel fuel additive, nanotechnology, nanotoxicology, consumer spray products  

    Progress and Final Reports:

    Original Abstract
  • 2011
  • 2012 Progress Report
  • 2013 Progress Report
  • 2014 Progress Report
  • 2015