Category Archives: risk

Nanoparticle behaviour in the environment unpredictable

These Swiss researchers took on a fairly massive project according to an April 19, 2017 news item on ScienceDaily,

The nanotech industry is booming. Every year, several thousands of tonnes of man-made nanoparticles are produced worldwide; sooner or later, a certain part of them will end up in bodies of water or soil. But even experts find it difficult to say exactly what happens to them there. It is a complex question, not only because there are many different types of man-made (engineered) nanoparticles, but also because the particles behave differently in the environment depending on the prevailing conditions.

Researchers led by Martin Scheringer, Senior Scientist at the Department of Chemistry and Applied Biosciences, wanted to bring some clarity to this issue. They reviewed 270 scientific studies, and the nearly 1,000 laboratory experiments described in them, looking for patterns in the behaviour of engineered nanoparticles. The goal was to make universal predictions about the behaviour of the particles.

An April 19, 2017ETH Zurich press release by Fabio Bergamin (also on EurekAlert), which originated the news item, elaborates,

Particles attach themselves to everything

However, the researchers found a very mixed picture when they looked at the data. “The situation is more complex than many scientists would previously have predicted,” says Scheringer. “We need to recognise that we can’t draw a uniform picture with the data available to us today.”

Nicole Sani-Kast, a doctoral student in Scheringer’s group and first author of the analysis published in the journal PNAS [Proceedings of the National Academy of Sciences], adds: “Engineered nanoparticles behave very dynamically and are highly reactive. They attach themselves to everything they find: to other nanoparticles in order to form agglomerates, or to other molecules present in the environment.”

Network analysis

To what exactly the particles react, and how quickly, depends on various factors such as the acidity of the water or soil, the concentration of the existing minerals and salts, and above all, the composition of the organic substances dissolved in the water or present in the soil. The fact that the engineered nanoparticles often have a surface coating makes things even more complicated. Depending on the environmental conditions, the particles retain or lose their coating, which in turn influences their reaction behaviour.

To evaluate the results available in the literature, Sani-Kast used a network analysis for the first time in this research field. It is a technique familiar in social research for measuring networks of social relations, and allowed her to show that the data available on engineered nanoparticles is inconsistent, insufficiently diverse and poorly structured.

More method for machine learning

“If more structured, consistent and sufficiently diverse data were available, it may be possible to discover universal patterns using machine learning methods,” says Scheringer, “but we’re not there yet.” Enough structured experimental data must first be available.

“In order for the scientific community to carry out such experiments in a systematic and standardised manner, some kind of coordination is necessary,” adds Sani-Kast, but she is aware that such work is difficult to coordinate. Scientists are generally well known for preferring to explore new methods and conditions rather than routinely performing standardized experiments.

[additional material]

Distinguishing man-made and natural nanoparticles

In addition to the lack of systematic research, there is also a second tangible problem in researching the behaviour of engineered nanoparticles: many engineered nanoparticles consist of chemical compounds that occur naturally in the soil. So far it has been difficult to measure the engineered particles in the environment since it is hard to distinguish them from naturally occurring particles with the same chemical composition.

However, researchers at ETH Zurich’s Department of Chemistry and Applied Biosciences, under the direction of ETH Professor Detlef Günther, have recently established an effective method that makes such a distinction possible in routine investigations. They used a state-of-the-art and highly sensitive mass spectrometry technique (called spICP-TOF mass spectrometry) to determine which chemical elements make up individual nanoparticles in a sample.

In collaboration with scientists from the University of Vienna, the ETH researchers applied the method to soil samples with natural cerium-containing particles, into which they mixed engineered cerium dioxide nanoparticles. Using machine learning methods, which were ideally suited to this particular issue, the researchers were able to identify differences in the chemical fingerprints of the two particle classes. “While artificially produced nanoparticles often consist of a single compound, natural nanoparticles usually still contain a number of additional chemical elements,” explains Alexander Gundlach-Graham, a postdoc in Günther’s group.

The new measuring method is very sensitive: the scientists were able to measure engineered particles in samples with up to one hundred times more natural particles.

The researchers have produced a visualization of their network analysis,

The researchers evaluated the experimental data published in the scientific literature using a network analysis. This analysis reveals which types of nanoparticles (blue) have been studied under which environmental conditions (red). (Visualisations: Thomas Kast)

Here are links and citation for two papers associated with this research,

A network perspective reveals decreasing material diversity in studies on nanoparticle interactions with dissolved organic matter by Nicole Sani-Kast, Jérôme Labille, Patrick Ollivier, Danielle Slomberg, Konrad Hungerbühler, and Martin Scheringer. PNAS 2017, 114: E1756-E1765, DOI: 10.1073/pnas.1608106114

Single-particle multi-element fingerprinting (spMEF) using inductively-coupled plasma time-of-flight mass spectrometry (ICP-TOFMS) to identify engineered nanoparticles against the elevated natural background in soils by Antonia Praetorius, Alexander Gundlach-Graham, Eli Goldberg, Willi Fabienke, Jana Navratilova, Andreas Gondikas, Ralf Kaegi, Detlef Günther, Thilo Hofmann, and Frank von der Kammer. Environonmental Science: Nano 2017, 4: 307-314, DOI: 10.1039/c6en00455e

Both papers are behind a paywall.

Is there a risk of resistance to nanosilver?

Anyone who’s noticed how popular silver has become as an antibacterial, antifungal, or antiviral agent may have wondered if resistance might occur as its use becomes more common. I have two bits on the topic, one from Australia and the other from Canada.

Australia

Researchers in Australia don’t have a definitive statement on the issue but are suggesting more caution (from a March 31, 2017 news item on Nanowerk),

Researchers at the University of Technology Sydney [UTS] warn that the broad-spectrum antimicrobial effectiveness of silver is being put at risk by the widespread and inappropriate expansion of nanosilver use in medical and consumer goods.

As well as their use in medical items such as wound dressings and catheters, silver nanoparticles are becoming ubiquitous in everyday items, including toothbrushes and toothpaste, baby bottles and teats, bedding, clothing and household appliances, because of their antibacterial potency and the incorrect assumption that ordinary items should be kept “clean” of microbes.

Nanobiologist Dr Cindy Gunawan, from the ithree institute at UTS and lead researcher on the investigation, said alarm bells should be ringing at the commercialisation of nanosilver use because of a “real threat” that resistance to nanosilver will develop and spread through microorganisms in the human body and the environment.

A March 31 (?), 2017 University of Technology Sydney press release by Fiona McGill, which originated the news item, expands on the theme,

Dr Gunawan and ithree institute director Professor Liz Harry, in collaboration with researchers at UNSW [University of New South Wales] and abroad, investigated more than 140 commercially available medical devices, including wound dressings and tracheal and urinary catheters, and dietary supplements, which are promoted as immunity boosters and consumed by throat or nasal spray.

Their perspective article in the journal ACS Nano concluded that the use of nanosilver in these items could lead to prolonged exposure to bioactive silver in the human body. Such exposure creates the conditions for microbial resistance to develop.

E. coli bacteria. Photo: Flickr/NIAID

 

The use of silver as an antimicrobial agent dates back centuries. Its ability to destroy pathogens while seemingly having low toxicity on human cells has seen it widely employed, in treating burns or purifying water, for example. More recently, ultra-small (less than 10,000th of a millimetre) silver nanoparticles have been engineered for antimicrobial purposes.  Their commercial appeal lies in superior potency at lower concentrations than “bulk” silver.

“Nanosilver is a proven antimicrobial agent whose reliability is being jeopardised by the commercialisation of people’s fear of bacteria,” Dr Gunawan said.

“Our use of it needs to be far more judicious, in the same way we need to approach antibiotic usage. Nanosilver is a useful tool but we need to be careful, use it wisely and only when the benefit outweighs the risk.

“People need to be made aware of just how widely it is used, but more importantly they need to be made aware that the presence of nanosilver has been shown to cause antimicrobial resistance.”

What is also needed, Dr Gunawan said, is a targeted surveillance strategy to monitor for any occurrence of resistance.

Professor Harry said the findings were a significant contribution to addressing the global antimicrobial resistance crisis.

“This research emphasises the threat posed to our health and that of the environment by the inappropriate use of nanosilver as an antibacterial, particularly in ordinary household and consumer items,” she said.

Here’s a link to and a citation for the paper,

Widespread and Indiscriminate Nanosilver Use: Genuine Potential for Microbial Resistance by Cindy Gunawan, Christopher P. Marquis, Rose Amal, Georgios A. Sotiriou, Scott A. Rice⊥, and Elizabeth J. Harry. ACS Nano, Article ASAP DOI: 10.1021/acsnano.7b01166 Publication Date (Web): March 24, 2017

Copyright © 2017 American Chemical Society

This paper is behind a paywall.

Meanwhile, researchers at the University Calgary (Alberta, Canada) may have discovered what could cause resistance to silver.

Canada

This April 25, 2017 news release on EurekAlert is from the Experimental Biology Annual Meeting 2017,

Silver and other metals have been used to fight infections since ancient times. Today, researchers are using sophisticated techniques such as the gene-editing platform Crispr-Cas9 to take a closer look at precisely how silver poisons pathogenic microbes–and when it fails. The work is yielding new insights on how to create effective antimicrobials and avoid the pitfalls of antimicrobial resistance.

Joe Lemire, a postdoctoral fellow at the University of Calgary, will present his work in this area at the American Society for Biochemistry and Molecular Biology annual meeting during the Experimental Biology 2017 meeting, to be held April 22-26 in Chicago.

“Our overarching goal is to deliver the relevant scientific evidence that would aid policymakers in developing guidelines for when and how silver could be used in the clinic to combat and control infectious pathogens,” said Lemire. “With our enhanced mechanistic understanding of silver toxicity, we also aim to develop novel silver-based antimicrobial therapies, and potentially rejuvenate other antibiotic therapies that bacteria have come to resist, via silver-based co-treatment strategies.”

Lemire and his colleagues are using Crispr-Cas9 genome editing to screen for and delete genes that allow certain bacterial species to resist silver’s antimicrobial properties. [emphasis mine] Although previous methods allowed researchers to identify genes that confer antibiotic resistance or tolerance, Crispr-Cas9 is the first technology to allow researchers to cleanly delete these genes from the genome without leaving behind any biochemical markers or “scars.”

The team has discovered many biological pathways involved in silver toxicity and some surprising ways that bacteria avoid succumbing to silver poisoning, Lemire said. While silver is used to control bacteria in many clinical settings and has been incorporated into hundreds of commercial products, gaining a more complete understanding of silver’s antimicrobial properties is necessary if we are to make the most of this ancient remedy for years to come.

###

Joe Lemire will present this research at 12-2:30 p.m. Tuesday, April 25, [2017] in Hall F, McCormick Place Convention Center (poster B379 939.2) (abstract). Contact the media team for more information or to obtain a free press pass to attend the meeting.

About Experimental Biology 2017

Experimental Biology is an annual meeting comprised of more than 14,000 scientists and exhibitors from six host societies and multiple guest societies. With a mission to share the newest scientific concepts and research findings shaping clinical advances, the meeting offers an unparalleled opportunity for exchange among scientists from across the U.S. and the world who represent dozens of scientific areas, from laboratory to translational to clinical research. http://www.experimentalbiology.org #expbio

About the American Society for Biochemistry and Molecular Biology (ASBMB)

ASBMB is a nonprofit scientific and educational organization with more than 12,000 members worldwide. Founded in 1906 to advance the science of biochemistry and molecular biology, the society publishes three peer-reviewed journals, advocates for funding of basic research and education, supports science education at all levels, and promotes the diversity of individuals entering the scientific workforce. http://www.asbmb.org

Lemire’s co-authors for the work presented at the 2017 annual meeting are: Kate Chatfield-Reed (The University of Calgary), Lindsay Kalan (Perelman School of Medicine), Natalie Gugala (The University of Calgary), Connor Westersund (The University of Calgary), Henrik Almblad (The University of Calgary), Gordon Chua (The University of Calgary), Raymond Turner (The University of Calgary).

For anyone who wants to pursue this research a little further, the most recent paper I can find is this one from 2015,

Silver oxynitrate: An Unexplored Silver Compound with Antimicrobial and Antibiofilm Activity by Joe A. Lemire, Lindsay Kalan, Alexandru Bradu, and Raymond J. Turner. Antimicrobial Agents and Chemotherapy 05177-14, doi: 10.1128/AAC.05177-14 Accepted manuscript posted online 27 April 2015

This paper appears to be open access.

European Commission has issued evaluation of nanomaterial risk frameworks and tools

Despite complaints that there should have been more, there has been some research into risks where nanomaterials are concerned. While additional research would be welcome, it’s perhaps more imperative that standardized testing and risk frameworks are developed so, for example, carbon nanotube safety research in Japan can be compared with the similar research in the Netherlands, the US, and elsewhere. This March 15, 2017 news item on Nanowerk features some research analyzing risk assessment frameworks and tools in Europe,

A recent study has evaluated frameworks and tools used in Europe to assess the potential health and environmental risks of manufactured nanomaterials. The study identifies a trend towards tools that provide protocols for conducting experiments, which enable more flexible and efficient hazard testing. Among its conclusions, however, it notes that no existing frameworks meet all the study’s evaluation criteria and calls for a new, more comprehensive framework.

A March 9, 2017 news alert in the European Commission’s Science for Environment Policy series, which originated the news item, provides more detail (Note: Links have been removed),

Nanotechnology is identified as a key emerging technology in the EU’s growth strategy, Europe 2020. It has great potential to contribute to innovation and economic growth and many of its applications have already received large investments. However,there are some uncertainties surrounding the environmental, health and safety risks of manufactured nanomaterials. For effective regulation, careful scientific analysis of their potential impacts is needed, as conducted through risk assessment exercises.

This study, conducted under the EU-funded MARINA project1, reviewed existing frameworks and tools for risk assessing manufactured nanomaterials. The researchers define a framework as a ‘conceptual paradigm’ of how a risk assessment should be conducted and understood, and give the REACH chemical safety assessment as an example. Tools are defined as implements used to carry out a specific task or function, such as experimental protocols, computer models or databases.

In all, 12 frameworks and 48 tools were evaluated. These were identified from other studies and projects. The frameworks were assessed against eight criteria which represent different strengths, such as whether they consider properties specific to nanomaterials, whether they consider the entire life cycle of a nanomaterial and whether they include careful planning and prioritise objectives before the risk assessment is conducted.

The tools were assessed against seven criteria, such as ease of use, whether they provide quantitative information and if they clearly communicate uncertainty in their results. The researchers defined the criteria for both frameworks and tools by reviewing other studies and by interviewing staff at organisations who develop tools.

The evaluation was thus able to produce a list of strengths and areas for improvement for the frameworks and tools, based on whether they meet each of the criteria. Among its many findings, the evaluation showed that most of the frameworks stress that ‘problem formulation’, which sets the goals and scope of an assessment during the planning process, is essential to avoid unnecessary testing. In addition, most frameworks consider routes of exposure in the initial stages of assessment, which is beneficial as it can exclude irrelevant exposure routes and avoid unnecessary tests.

However, none of the frameworks met all eight of the criteria. The study therefore recommends that a new, comprehensive framework is developed that meets all criteria. Such a framework is needed to inform regulation, the researchers say, and should integrate human health and environmental factors, and cover all stages of the life cycle of a product containing nanomaterials.

The evaluation of the tools suggested that many of them are designed to screen risks, and not necessarily to support regulatory risk assessment. However, their strengths include a growing trend in quantitative models, which can assess uncertainty; for example, one tool analysed can identify uncertainties in its results that are due to gaps in knowledge about a material’s origin, characteristics and use.

The researchers also identified a growing trend in tools that provide protocols for experiments, such as identifying materials and test hazards, which are reproducible across laboratories. These tools could lead to a shift from expensive case-by-case testing for risk assessment of manufactured nanomaterials towards a more efficient process based on groupings of nanomaterials; and ‘read-across’ methods, where the properties of one material can be inferred without testing, based on the known properties of a similar material. The researchers do note, however, that although read-across methods are well established for chemical substances, they are still being developed for nanomaterials. To improve nanomaterial read-across methods, they suggest that more data are needed on the links between nanomaterials’ specific properties and their biological effects.

That’s all, folks.

The Swiss come to a better understanding of nanomaterials

Just to keep things interesting, after the report suggesting most of the information that the OECD (Organization for Economic Cooperation and Development) has on nanomaterials is of little value for determining risk (see my April 5, 2017 posting for more) the Swiss government has released a report where they claim an improved understanding of nanomaterials than they previously had due to further research into the matter. From an April 6, 2017 news item on Nanowerk,

In the past six years, the [Swiss] National Research Programme “Opportunities and Risks of Nanomaterials” (NRP 64) intensively studied the development, use, behaviour and degradation of engineered nanomaterials, including their impact on humans and on the environment.

Twenty-three research projects on biomedicine, the environment, energy, construction materials and food demonstrated the enormous potential of engineered nanoparticles for numerous applications in industry and medicine. Thanks to these projects we now know a great deal more about the risks associated with nanomaterials and are therefore able to more accurately determine where and how they can be safely used.

An April 6, 2017 Swiss National Science Foundation press release, which originated the news item, expands on the theme,

“One of the specified criteria in the programme was that every project had to examine both the opportunities and the risks, and in some cases this was a major challenge for the researchers,” explains Peter Gehr, President of the NRP 64 Steering Committee.

One development that is nearing industrial application concerns a building material strengthened with nanocellulose that can be used to produce a strong but lightweight insulation material. Successful research was also carried out in the area of energy, where the aim was to find a way to make lithium-ion batteries safer and more efficient.

Promising outlook for nanomedicine

A great deal of potential is predicted for the field of nanomedicine. Nine of the 23 projects in NRP 64 focused on biomedical applications of nanoparticles. These include their use for drug delivery, for example in the fight against viruses, or as immune modulators in a vaccine against asthma. Another promising application concerns the use of nanomagnets for filtering out harmful metallic substances from the blood. One of the projects demonstrated that certain nanoparticles can penetrate the placenta barrier, which points to potential new therapy options. The potential of cartilage and bone substitute materials based on nanocellulose or nanofibres was also studied.

The examination of potential health risks was the focus of NRP 64. A number of projects examined what happens when nanoparticles are inhaled, while two focused on ingestion. One of these investigated whether the human gut is able to absorb iron more efficiently if it is administered in the form of iron nanoparticles in a food additive, while the other studied silicon nanoparticles as they occur in powdered condiments. It was ascertained that further studies will be required in order to determine the doses that can be used without risking an inflammatory reaction in the gut.

What happens to engineered nanomaterials in the environment?

The aim of the seven projects focusing on environmental impact was to gain a better understanding of the toxicity of nanomaterials and their degradability, stability and accumulation in the environment and in biological systems. Here, the research teams monitored how engineered nanoparticles disseminate along their lifecycle, and where they end up or how they can be discarded.

One of the projects established that 95 per cent of silver nanoparticles that are washed out of textiles are collected in sewage treatment plants, while the remaining particles end up in sewage sludge, which in Switzerland is incinerated. In another project a measurement device was developed to determine how aquatic microorganisms react when they come into contact with nanoparticles.

Applying results and making them available to industry

“The findings of the NRP 64 projects form the basis for a safe application of nanomaterials,” says Christoph Studer from the Federal Office of Public Health. “It has become apparent that regulatory instruments such as testing guidelines will have to be adapted at both national and international level.” Studer has been closely monitoring the research programme in his capacity as the Swiss government’s representative in NRP 64. In this context, the precautionary matrix developed by the government is an important instrument by means of which companies can systematically assess the risks associated with the use of nanomaterials in their production processes.

The importance of standardised characterisation and evaluation of engineered nanomaterials was highlighted by the close cooperation among researchers in the programme. “The research network that was built up in the framework of NRP 64 is functioning smoothly and needs to be further nurtured,” says Professor Bernd Nowack from Empa, who headed one of the 23 projects.

The results of NRP 64 show that new key technologies such as the use of nanomaterials need to be closely monitored through basic research due to the lack of data on its long-term effects. As Peter Gehr points out, “We now know a lot more about the risks of nanomaterials and how to keep them under control. However, we need to conduct additional research to learn what happens when humans and the environment are exposed to engineered nanoparticles over longer periods, or what happens a long time after a one-off exposure.”

You can find out more about the Opportunities and Risks of Nanomaterials; National Research Programme (NRP 64) here.

OECD (Organization for Economic Cooperation and Development) Dossiers on Nanomaterials Are of “Little to No Value for assessing risk?”

The announcement that a significant portion of the OECD’s (Organization for Economic Cooperation and Development) dossiers on 11 nanomaterials have next to no value for assessing risk seems a harsh judgment from the Center for International Environmental Law (CIEL). From a March 1, 2017 posting by Lynn L. Bergeson on the Nanotechnology Now,

On February 23, 2017, the Center for International Environmental Law (CIEL) issued a press release announcing a new report, commissioned by CIEL, the European Environmental Citizens’ Organization for Standardization (ECOS), and the Oeko-Institute, that “shows that most of the information made available by the Sponsorship Testing Programme of the Organisation for Economic Co-operation and Development (OECD) is of little to no value for the regulatory risk assessment of nanomaterials.”

Here’s more from the Feb. 23, 3017 CIEL press release, which originated the posting,

The study published today [Feb. 23, 2017] was delivered by the Institute of Occupational Medicine (IOM) based in Singapore. IOM screened the 11,500 pages of raw data of the OECD dossiers on 11 nanomaterials, and analysed all characterisation and toxicity data on three specific nanomaterials – fullerenes, single-walled carbon nanotubes, and zinc oxide.

“EU policy makers and industry are using the existence of the data to dispel concerns about the potential health and environmental risks of manufactured nanomaterials,” said David Azoulay, Senior Attorney for CIEL. “When you analyse the data, in most cases, it is impossible to assess what material was actually tested. The fact that data exists about a nanomaterial does not mean that the information is reliable to assess the hazards or risks of the material.”

The dossiers were published in 2015 by the OECD’s Working Party on Manufactured Nanomaterials (WPMN), which has yet to draw conclusions on the data quality. Despite this missing analysis, some stakeholders participating in EU policy-making – notably the European Chemicals Agency (ECHA) and the European Commission’s Joint Research Centre – have presented the dossiers as containing information on nano-specific human health and environmental impacts. Industry federations and individual companies have taken this a step further emphasizing that there is enough information available to discard most concerns about potential health or environmental risks of manufactured nanomaterials.

“Our study shows these claims that there is sufficient data available on nanomaterials are not only false, but dangerously so,” said Doreen Fedrigo, Senior Policy Officer of ECOS. ”The lack of nano-specific information in the dossiers means that the results of the tests cannot be used as evidence of no ‘nano-effect’ of the tested material. This information is crucial for regulators and producers who need to know the hazard profile of these materials. Analysing the dossiers has shown that legislation detailing nano-specific information requirements is crucial for the regulatory risk assessment of nanomaterials.”

The report provides important recommendations on future steps in the governance of nanomaterials. “Based on our analysis, serious gaps in current dossiers must be filled in with characterisation information, preparation protocols, and exposure data,” said Andreas Hermann of the Oeko-Institute. “Using these dossiers as they are and ignoring these recommendations would mean making decisions on the safety of nanomaterials based on faulty and incomplete data. Our health and environment requires more from producers and regulators.”

CIEL has an Analysis of OECD WPMN Dossiers Regarding the Availability of Data to Evaluate and Regulate Risk (Dec 2016) webpage which provides more information about the dossiers and about the research into the dossiers and includes links to the report, the executive summer, and the dataset,

The Sponsorship Testing Programme of the Working Party on Manufactured Nanomaterials (WPMN) of the Organisation for Economic Co-operation and Development (OECD) started in 2007 with the aim to test a selection of 13 representative nanomaterials for many endpoints. The main objectives of the programme were to better understand what information on intrinsic properties of the nanomaterials might be relevant for exposure and hazards assessment and assess the validity of OECD chemicals Test Guidelines for nanomaterials. The testing programme concluded in 2015 with the publication of dossiers on 11 nanomaterials: 11,500 pages of raw data to be analysed and interpreted.

The WPMN has not drawn conclusions on the data quality, but some stakeholders participating in EU policy-making – notably the European Chemicals Agency and the European Commission’s Joint Research Centre – presented the dossiers as containing much scientific information that provided a better understanding of their nano-specific human health and environmental impacts. Industry federations and individual companies echoed the views, highlighting that there was enough information available to discard most concerns about potential health or environmental risks of manufactured nanomaterials.

As for the OECD, it concluded, even before the publication of the dossiers, that “many of the existing guidelines are also suitable for the safety assessment of nanomaterials” and “the outcomes (of the sponsorship programme) will provide useful information on the ‘intrinsic properties’ of nanomaterials.”

The Center for International Environmental Law (CIEL), the European Citizens’ Organisation for Standardisation (ECOS) and the Öko-Institut commissioned scientific analysis of these dossiers to assess the relevance of the data for regulatory risk assessment.

The resulting report: Analysis of OECD WPMN dossiers regarding the availability of data to evaluate and regulate risk, provides insights illustratating how most of the information made available by the sponsorship programme is of little to no value in identifying hazards or in assessing risks due to nanomaterials.

The analysis shows that:

  • Most studies and documents in the dossiers contain insufficient characterisation data about the specific nanomaterial addressed (size, particle distribution, surface shape, etc.), making it impossible to assess what material was actually tested.
  • This makes it impossible to make any firm statements regarding the nano-specificity of the hazard data published, or the relationship between observed effects and specific nano-scale properties.
  • Less than 2% of the study records provide detail on the size of the nanomaterial tested. Most studies use mass rather than number or size distribution (so not following scientifically recommended reporting practice).
  • The absence of details on the method used to prepare the nanomaterial makes it virtually impossible to correlate an identified hazard with specific nanomaterial characteristic. Since the studies do not indicate dispersion protocols used, it is impossible to assess whether the final dispersion contained the intended mass concentration (or even the actual presence of nanomaterials in the test system), how much agglomeration may have occurred, and how the preparation protocols may have influenced the size distribution.
  • There is not enough nano-specific information in the dossiers to inform about nano-characteristics of the raw material that influence their toxicology. This information is important for regulators and its absence makes information in the dossier irrelevant to develop read-across guidelines.
  • Only about half of the endpoint study records using OECD Test Guideliness (TGs) were delivered using unaltered OECD TGs, thereby respecting the Guidelines’ requirements. The reasons for modifications of the TGs used in the tests are not clear from the documentation. This includes whether the study record was modified to account for challenges related to specific nanomaterial properties or for other, non-nano-specific reasons.
  • The studies do not contain systematic testing of the influence of nano-specific characteristics on the study outcome, and they do not provide the data needed to assess the effect of nano-scale features on the Test Guidelines. Given the absence of fundamental information on nanomaterial characteristics, the dossiers do not provide evidence of the applicability of existing OECD Test Guidelines to nanomaterials.

The analysis therefore dispels several myths created by some stakeholders following publication of the dossiers and provides important perspective for the governance of nanomaterials. In particular, the analysis makes recommendations to:

  • Systematically assess the validity of existing Test Guidelines for relevance to nanomaterials
  • Develop Test Guidelines for dispersion and other test preparations
  • Define the minimum characteristics of nanomaterials that need to be reported
  • Support the build-up of exposure database
  • Fill the gaps in current dossiers with characterisation information, preparation protocols and exposure data

Read full report.
Read executive summary.
Download full dataset.

This is not my area of expertise and while I find the language a bit inflammatory, it’s my understanding that there are great gaps in our understanding of nanomaterials and testing for risk assessment has been criticized for many of the reasons pointed out by CIEL, ECOS, and the Oeko-Institute.

You can find out more about CIEL here; ECOS here; and the Oeko-Institute (also known as Öko-Institute) here.

Findings on oral exposure to nanoscale titanium dioxide

It’s been a while since I’ve run a piece on health concerns and nanoparticles. The nanoparticles in question are titanium dioxide and the concerns centre on oral exposure to them according to a Jan. 24, 2017 news item on Nanowerk,

Researchers from INRA [French National Institute for Agricultural Research] and their partners have studied the effects of oral exposure to titanium dioxide, an additive (E171) commonly used in foodstuffs, especially confectionary. They have shown for the first time that E171 crosses the intestinal barrier in animals and reaches other parts of the body.

Immune system disorders linked to the absorption of the nanoscale fraction of E171 particles were observed. The researchers also showed that chronic oral exposure to the additive spontaneously induced preneoplastic lesions in the colon, a non-malignant stage of carcinogenesis, in 40% of exposed animals.

Moreover, E171 was found to accelerate the development of lesions previously induced for experimental purposes. While the findings show that the additive plays a role in initiating and promoting the early stages of colorectal carcinogenesis, they cannot be extrapolated to humans or more advanced stages of the disease. [emphasis mine]

A Jan. 20, 2017 IINRA press release, which originated the news item,  provides more detail about European use of titanium dioxide as a food additive and about the research,

Present in many products including cosmetics, sunscreens, paint and building materials, titanium dioxide (or TiO2), known as E171 in Europe, is also widely used as an additive in the food industry to whiten or give opacity to products. It is commonly found in sweets, chocolate products, biscuits, chewing gum and food supplements, as well as in toothpaste and pharmaceutical products. Composed of micro- and nanoparticles, E171 is nevertheless not labelled a “nanomaterial”, since it does not contain more than 50% of nanoparticles (in general it contains from 10-40%). The International Agency for Research on Cancer (IARC) evaluated the risk of exposure to titanium dioxide by inhalation (occupational exposure), resulting in a Group 2B classification, reserved for potential carcinogens for humans.

Today, oral exposure to E171 is a concern, especially in children who tend to eat a lot of sweets. INRA researchers studied the product as a whole (that is, its mixed composition of micro- and nanoparticules), and have also evaluated the effect of the nanoscale particle fraction alone, by comparing it to a model nanoparticle.

Titanium dioxide crosses the intestinal barrier and passes into the bloodstream

The researchers exposed rats orally to a dose of 10mg of E171 per kilogram of body weight per day, similar to the exposure humans experience through food consumption (data from European Food Safety Agency, September 20162). They showed for the first time in vivo that titanium dioxide is absorbed by the intestine and passes into the bloodstream. Indeed, the researchers found titanium dioxide particles in the animals’ livers.

Titanium dioxide alters intestinal and systemic immune response

Titanium dioxide nanoparticles were present in the lining of the small intestine and in the colon, and entered the nuclei of the immune cells of Peyer’s patches, which induce immune response in the intestine. The researchers showed an imbalance in immune response, ranging from a defect in the production of cytokines in Peyer’s patches to the development of micro-inflammation in colon mucosa. In the spleen, representative of systemic immunity, exposure to E171 increases the capacity of immune cells to produce pro-inflammatory cytokines when they are activated in vitro.

Chronic oral exposure to titanium dioxide plays a role in initiating and promoting early stages of colorectal carcinogenesis

The researchers exposed rats to regular oral doses of titanium dioxide through drinking water for 100 days. In a group of rats previously treated with an experimental carcinogen, exposure to TiO2 led to an increase in the size of preneoplastic lesions. In a group of healthy rats exposed to E171, four out of eleven spontaneously developed preneoplastic lesions in the intestinal epithelium. Non-exposed animals presented no anomalies at the end of the 100-day study. These results indicate that E171 both initiates and promotes the early stages of colorectal carcinogenesis in animals.

These studies show for the first time that the additive E171 is a source of titanium dioxide nanoparticles in the intestine and the entire body, with consequences for both immune function and the development of preneoplastic lesions in the colon. These first findings justify a carcinogenesis study carried out under OECD [Organization for Economic Cooperation and Development] guidelines to continue observations at a later stage of cancer. They provide new data for evaluating the risks of the E171 additive in humans.

These studies were carried out within the framework of the Nanogut project, financed by the French Agency for Food, Environmental and Occupational Health & Safety (ANSES) within the French national programme for research related to the environment, health and the workplace (PNR EST) and coordinated by INRA. Sarah Bettini’s university thesis contract was financed by the French laboratory of excellence LabEx SERENADE.

Here’s a link to and a citation for the paper,

Food-grade TiO2 impairs intestinal and systemic immune homeostasis, initiates preneoplastic lesions and promotes aberrant crypt development in the rat colon by Sarah Bettini, Elisa Boutet-Robinet, Christel Cartier, Christine Coméra, Eric Gaultier, Jacques Dupuy, Nathalie Naud, Sylviane Taché, Patrick Grysan, Solenn Reguer, Nathalie Thieriet, Matthieu Réfrégiers, Dominique Thiaudière, Jean-Pierre Cravedi, Marie Carrière, Jean-Nicolas Audinot, Fabrice H. Pierre, Laurence Guzylack-Piriou, & Eric Houdeau. Scientific Reports 7, Article number: 40373 (2017) doi:10.1038/srep40373 Published online: 20 January 2017

This paper is open access.

The research is concerning but they don’t want to draw any conclusions yet, which explains the recommendation for further research.

Using sugar for a better way to clean nanoparticles from organisms

Researchers at the US National Institute of Standards and Technology (NIST) have found that a laboratory technique used for over 60 years is the best way to date to clean nanoparticles from organisms. From a Jan. 26, 2017 news item on ScienceDaily,

Sometimes old-school methods provide the best ways of studying cutting-edge tech and its effects on the modern world.

Giving a 65-year-old laboratory technique a new role, researchers at the National Institute of Standards and Technology (NIST) have performed the cleanest separation to date of synthetic nanoparticles from a living organism. The new NIST method is expected to significantly improve experiments looking at the potential environmental and health impacts of these manufactured entities. It will allow scientists to more accurately count how many nanoparticles have actually been ingested by organisms exposed to them.

A Jan. 26, 2017 NIST news release (also on EurekAlert), which originated the news item, offers more detail,

The common roundworm Caenorhabditis elegans has been used in recent years as a living model for laboratory studies of how biological and chemical compounds may affect multicellular organisms. These compounds include engineered nanoparticles (ENPs), bits of material between 1 and 100 nanometers (billionths of a meter, or about 1/10,000 the diameter of a red blood cell). Previous research has often focused on quantifying the amount and size of engineered nanoparticles ingested by C. elegans. Measuring the nanoparticles that actually make it into an organism is considered a more relevant indicator of potential toxicity than just the amount of ENPs to which the worms are exposed.

Traditional methods for counting ingested ENPs have produced questionable results. Currently, researchers expose C. elegans to metal ENPs such as silver or gold in solution, then rinse the excess particles away with water followed by centrifugation and freeze-drying. A portion of the “cleaned” sample produced is then typically examined by a technique that determines the amount of metal present, known as inductively coupled plasma mass spectrometry (ICP-MS). It often yields ENP counts in the tens of thousands per worm; however, those numbers always seem too high to NIST researchers working with C. elegans.

“Since ICP-MS will detect all of the nanoparticles associated with the worms, both those ingested and those that remain attached externally, we suspect that the latter is what makes the ‘ENPs’ per-worm counts so high,” said NIST analytical chemist Monique Johnson (link sends e-mail), the lead author on the ACS Nano paper. “Since we only wanted to quantify the ingested ENPs, a more robust and reliable separation method was needed.”

Luckily, the solution to the problem was already in the lab.

Cross section of the roundworm C. elegans

Scanning electron micrograph showing a cross section of the roundworm C. elegans with two ingested engineered nanoparticles (red dots just right of center). Images such as this provided NIST researchers with visual confirmation that nanoparticle consumption actually occurred. Credit: K. Scott/NIST

In the course of culturing C. elegans for ENP-exposure experiments, Johnson and her colleagues had used sucrose density gradient centrifugation, a decades-old and established system for cleanly separating cellular components, to isolate the worms from debris and bacteria. “We wondered if the same process would allow us to perform an organism-from-ENP separation as well, so I designed a study to find out,” Johnson said.

In their experiment, the NIST researchers first exposed separate samples of C. elegans to low and high concentrations of two sizes of gold nanospheres, 30 and 60 nanometers in diameter. The researchers put each of the samples into a centrifuge and removed the supernatant (liquid portion), leaving the worms and ENPs in the remaining pellets. These were centrifuged twice in a salt solution (rather than just water as in previous separation methods), and then centrifuged again, but this time, through a uniquely designed sucrose density gradient.

“From top to bottom, our gradient consisted of a salt solution layer to trap excess ENPs and three increasingly dense layers of sucrose [20, 40 and 50 percent] to isolate the C. elegans,” Johnson explained. “We followed up the gradient with three water rinses and with centrifugations to ensure that only worms with ingested ENPs, and not the sucrose separation medium with any excess ENPs, would make it into the final pellet.”

Analyzing the range of masses in the ultrapurified samples indicated gold levels more in line with what the researchers expected would be found as ingested ENPs. Experimental validation of the NIST separation method’s success came when the worms were examined in detail under a scanning electron microscope (SEM).

“For me, the eureka moment was when I first saw gold ENPs in the cross section images taken from the C. elegans samples that had been processed through the sucrose density gradient,” Johnson said. “I had been dreaming about finding ENPs in the worm’s digestive tract and now they were really there!”

The high-resolution SEM images also provided visual evidence that only ingested ENPs were counted. “No ENPs were attached to the cuticle, the exoskeleton of C. elegans, in any of the sucrose density gradient samples,” Johnson said. “When we examined worms from our control experiments [processed using the traditional no-gradient, water-rinse-only separation method], there were a number of nanospheres found attached to the cuticle.

Now that it has been successfully demonstrated, the NIST researchers plan to refine and further validate their system for evaluating the uptake of ENPs by C. elegans. “Hopefully, our method will become a useful and valuable tool for reducing the measurement variability and sampling bias that can plague environmental nanotoxicology studies,” Johnson said.

They’ve tested this technique on gold nanoparticles, which begs the question, What kinds of nanoparticles can this technique be used for? Metal nanoparticles only or all nanoparticles?

I’m sure the researchers have already asked these questions and started researching the answers. While the rest of us wait, here’s a link to and a citation for the paper about this promising new technique,

Separation, Sizing, and Quantitation of Engineered Nanoparticles in an Organism Model Using Inductively Coupled Plasma Mass Spectrometry and Image Analysis by Monique E. Johnson, Shannon K. Hanna, Antonio R. Montoro Bustos, Christopher M. Sims, Lindsay C. C. Elliott, Akshay Lingayat, Adrian C. Johnston, Babak Nikoobakht, John T. Elliott, R. David Holbrook, Keana C. K. Scott, Karen E. Murphy, Elijah J. Petersen, Lee L. Yu, and Bryant C. Nelson. ACS Nano, 2017, 11 (1), pp 526–540 DOI: 10.1021/acsnano.6b06582 Publication Date (Web): December 16, 2016

Copyright This article not subject to U.S. Copyright. Published 2016 by the American Chemical Society

This paper is behind a paywall.

Nanoparticles can activate viruses lying dormant in lung cells

The nanoparticles in question are from combustion engines, which means that we are exposed to them. One other note, the testing has not been done on humans but rather on cells. From a Jan. 16, 2017 news item on ScienceDaily,

Nanoparticles from combustion engines can activate viruses that are dormant in in lung tissue cells. This is the result of a study by researchers of Helmholtz Zentrum München, a partner in the German Center for Lung Research (DZL), which has now been published in the journal Particle and Fibre Toxicology.

To evade the immune system, some viruses hide in cells of their host and persist there. In medical terminology, this state is referred to as a latent infection. If the immune system becomes weakened or if certain conditions change, the viruses become active again, begin to proliferate and destroy the host cell. A team of scientists led by Dr. Tobias Stöger of the Institute of Lung Biology and Prof. Dr. Heiko Adler, deputy head of the research unit Lung Repair and Regeneration at Helmholtz Zentrum München, now report that nanoparticles can also trigger this process.

A Jan. 16, 2017 Helmholtz Zentrum München press release (also on EurekAlert), which originated the news item, provides more detail,

“From previous model studies we already knew that the inhalation of nanoparticles has an inflammatory effect and alters the immune system,” said study leader Stöger. Together with his colleagues Heiko Adler and Prof. Dr. Philippe Schmitt-Kopplin, he showed that “an exposure to nanoparticles can reactivate latent herpes viruses in the lung.”

Specifically, the scientists tested the influence of nanoparticles typically generated by fossil fuel combustion in an experimental model for a particular herpes virus infection. They detected a significant increase in viral proteins, which are only produced with active virus proliferation. “Metabolic and gene expression analyses also revealed patterns resembling acute infection,” said Philippe Schmitt-Kopplin, head of the research unit Analytical BioGeoChemistry (BGC). Moreover, further experiments with human cells demonstrated that Epstein-Barr viruses are also ‘awakened’ when they come into contact with the nanoparticles.

Potential approach for chronic lung diseases

In further studies, the research team would like to test whether the results can also be transferred to humans. “Many people carry herpes viruses, and patients with idiopathic pulmonary fibrosis are particularly affected,” said Heiko Adler. “If the results are confirmed in humans, it would be important to investigate the molecular process of the reactivation of latent herpes viruses induced by particle inhalation. Then we could try to influence this pathway therapeutically.”

Special cell culture models shall therefore elucidate the exact mechanism of virus reactivation by nanoparticles. “In addition,” Stöger said, ”in long-term studies we would like to investigate to what extent  repeated nanoparticle exposure with corresponding virus reactivation leads to chronic inflammatory and remodeling processes in the lung.”

Further Information

Background:
In 2015 another group at the Helmholtz Zentrum München demonstrated how the Epstein-Barr virus  hides in human cells. In March 2016 researchers also showed that microRNAs silence immune alarm signals of cells infected with the Epstein-Barr virus.

Original Publication:
Sattler, C. et al. (2016): Nanoparticle exposure reactivates latent herpesvirus and restores a signature of acute infection. Particle and Fibre Toxicology, DOI 10.1186/s12989-016-0181-1

Here’s a link to and a citation for the paper on investigating latent herpes virus,

Nanoparticle exposure reactivates latent herpesvirus and restores a signature of acute infection by Christine Sattler, Franco Moritz, Shanze Chen, Beatrix Steer, David Kutschke, Martin Irmler, Johannes Beckers, Oliver Eickelberg, Philippe Schmitt-Kopplin, Heiko Adler. Particle and Fibre Toxicology201714:2 DOI: 10.1186/s12989-016-0181-1 Published: 10 January 2017

©  The Author(s). 2017

This paper is open access and, so too, is the 2016 paper.

2016 report on nanomaterial reporting released by French government

Lynn L. Bergeson has announced the release of a new report from the French government in her Jan. 3, 2017 posting on Nanotechnology Now,

In November 2016, the Ministry of the Environment, Energy, and the Sea released its 2016 report, in French, Éléments issus des déclarations des substances à l’état nanoparticulaire. … The report analyzes nanomaterial declarations received in 2016 for reporting year 2015. Under Decree No. 2012-232, companies that manufacture, import, and/or distribute a “substance with nanoparticle status” in an amount of at least 100 grams per year must submit an annual report with substance identity, quantity, and use information. According to the report, while the number of declarations received in 2016 decreased from 2015, the quantity of materials produced increased (350,487 tonnes vs. 300,822 tonnes in 2015), as well as the quantity imported (125,279 tonnes vs. 114,951 tonnes in 2015).

For people with the French language skills, you can find the report here (PDF). You can also check out the R-Nano website (French language) (English language) for more information about the reporting programme in France.

In related news. the US Environmental Protection Agency announced its one-time only nanomaterial reporting requirements as highlighted in my Jan. 18, 2017 posting.

US Environmental Protection Agency finalizes its one-time reporting requirements for nanomaterials

The US Environmental Protection Agency (EPA) has announced its one-time reporting requirement for  nanomaterials. From a Jan. 12, 2017 news item on Nanowerk,

The U.S. Environmental Protection Agency (EPA) is requiring one-time reporting and recordkeeping requirements on nanoscale chemical substances in the marketplace. These substances are nano-sized versions of chemicals that are already in the marketplace.
EPA seeks to facilitate innovation while ensuring safety of the substances. EPA currently reviews new chemical substances manufactured or processed as nanomaterials prior to introduction into the marketplace to ensure that they are safe.

For the first time, EPA is using [the] TSCA [Toxic Substances Control Act] to collect existing exposure and health and safety information on chemicals currently in the marketplace when manufactured or processed as nanoscale materials.

The companies will notify EPA of certain information:
– specific chemical identity;
– production volume;
– methods of manufacture; processing, use, exposure, and release information; and,available health and safety data.

Reactions

David Stegon writes about the requirement in a Jan. 12, 2017 posting on Chemical Watch,

The US EPA has finalised its nanoscale materials reporting rule, completing a process that began more than 11 years ago.

The US position contrasts with that of the European Commission, which has rejected the idea of a specific mandatory reporting obligation for nanomaterials. Instead it insists such data can be collected under REACH’s registration rules for substances in general. It has told Echa [ECHA {European Chemicals Agency}] to develop ‘nano observatory’ pages on its website with existing nanomaterial information. Meanwhile, Canada set its reporting requirements in 2015.

The US rule, which comes under section 8(a) of TSCA, will take effect 120 days after publication in the Federal Register.

It defines nanomaterials as chemical substances that are:

  • solids at 25 degrees Celsius at standard atmospheric pressure;
  • manufactured or processed in a form where any particles, including aggregates and agglomerates, are between 1 and 100 nanometers (nm) in at least one dimension; and
  • manufactured or processed to exhibit one or more unique and novel property.

The rule does not apply to chemical substances manufactured or processed in forms that contain less than 1% by weight of any particles between 1 and 100nm.

Taking account of comments received on the rulemaking, the EPA made three changes to the proposed definition:

  • it added the definition of unique and novel properties to help identify substances that act differently at nano sizes;
  • it clarified that a substance is not a nanomaterial if it fits the specified size range, but does not have a size-dependent property that differs from the same chemical at sizes greater than 100nm; and
  • it eliminated part of the nanomaterial definition that had said a reportable chemical may not include a substance that only has trace amounts of primary particles, aggregates, or agglomerates in the size range of 1 to 100nm.

The EPA has added the new information gathering rule (scroll down about 50% of the way) on its Control of Nanoscale Materials under the Toxic Substances Control Act webpage.

There’s also this Jan. 17, 2017 article by Meagan Parrish for the ChemInfo which provides an alternative perspective and includes what appears to be some misinformation (Note: A link has been removed),

It was several years in the making, but in the final stages of its rule-making process for nanomaterial reporting, the Environmental Protection Agency declined to consider feedback from the industry.

Now, with the final language published and the rule set to go into effect in May, some in the industry are concerned that the agency is requiring an unnecessary amount of costly reporting that isn’t likely to reveal potential hazards. The heightened regulations could also hamper the pace of innovation underway in the industry.

“The poster child for nanotechnology is carbon nanotubes,” says James Votaw, a partner with Manatt, Phelps & Phillips, of the form of carbon that is 10,000 smaller than human hair but stronger than steel. “It can be used to make very strong materials and as an additive in plastics to make them electrically conductive or stiffer.”

The EPA has been attempting to define nanomaterials since 2004 and assess the potential for environmental or human health risks associated with their use. In 2008, the EPA launched an effort to collect voluntarily submitted information from key players in the industry, but after a few years, the agency wasn’t happy with amount of responses. The effort to create a mandatory reporting requirement was launched in 2010.

Yet, according to Votaw, after a 2015 proposal of the rule was extensively criticized by the industry for being overly ambiguous and overly inclusive of its coverage, the industry asked the EPA to reopen a dialogue on the rule. The EPA declined.

The new reporting requirement is expected to cost companies about $27.79 million during the first year and $3.09 million in subsequent years. [emphasis mine]

As far as I’m aware, this is a one-time reporting requirement. Although I’m sure many would like to see that change.

As for the Canadian situation, I mentioned the nanomaterials mandatory survey noted in Stegon’s piece in a July 29, 2015 posting. It was one of a series of mandatory surveys (currently, a survey on asbestos is underway) issued as part of Canada’s Chemicals Management Plan. You can find more information about the nanomaterials notice and approach to the survey although there doesn’t appear to have been a report made public but perhaps it’s too soon. From the Nanomaterials Mandatory Survey page,

The Government of Canada is undertaking a stepwise approach to address nanoscale forms of substances on the DSL. The proposed approach consists of three phases:

  • Establishment of a list of existing nanomaterials in Canada (this includes the section 71 Notice);
  • Prioritization of existing nanomaterials for action; and
  • Action on substances identified for further work.

The overall approach was first described in a consultation document entitled Proposed Approach to Address Nanoscale Forms of Substances on the Domestic Substances List, published on March 18, 2015. This consultation document was open for a 60-day public comment period to solicit feedback from stakeholders, particularly on the first phase of the approach.

A second consultation document entitled Proposed Prioritization Approach for Nanoscale Forms of Substances on the Domestic Substances List was published on July 27, 2016. In this document, the approach proposed for prioritization of existing nanomaterials on the DSL is described, taking into consideration the results of the section 71 Notice.  Comments on this consultation document may be submitted prior to September 25, 2016 …

I look forward to discovering a report on the Canadian nanomaterials survey should one be made public.