Tag Archives: nanotoxicology

New method for measuring risks and quantities of engineered nanomaterials delivered to cells

Despite all the talk about testing engineered nanoparticles and their possible effects on cells, there are problems with the testing process which researchers at the Harvard School of Public Health (HSPH) claim to have addressed (h/t Nanowerk, March 28, 2014).

A March 28, 2014 HSPH press release explains the interest in testing the effects of engineered nanomaterials/nanoparticles on health and describes some of the problems associated with testing their interaction with cells,

Thousands of consumer products containing engineered nanoparticles — microscopic particles found in everyday items from cosmetics and clothing to building materials — enter the market every year. Concerns about possible environmental health and safety issues of these nano-enabled products continue to grow with scientists struggling to come up with fast, cheap, and easy-to-use cellular screening systems to determine possible hazards of vast libraries of engineered nanomaterials. However, determining how much exposure to engineered nanoparticles could be unsafe for humans requires precise knowledge of the amount (dose) of nanomaterials interacting with cells and tissues such as lungs and skin.

With chemicals, this is easy to do but when it comes to nanoparticles suspended in physiological media, this is not trivial. Engineered nanoparticles in biological media interact with serum proteins and form larger agglomerates which alter both their so called effective density and active surface area and ultimately define their delivery to cell dose and bio-interactions. This behavior has tremendous implications not only in measuring the exact amount of nanomaterials interacting with cells and tissue but also in defining hazard rankings of various engineered nanomaterials (ENMs). As a result, thousands of published cellular screening assays are difficult to interpret and use for risk assessment purposes.

The press release goes on to describe the new technique (Note: Links have been removed),

Scientists at the Center for Nanotechnology and Nanotoxicology at Harvard School of Public Health (HSPH) have discovered a fast, simple, and inexpensive method to measure the effective density of engineered nanoparticles in physiological fluids, thereby making it possible to accurately determine the amount of nanomaterials that come into contact with cells and tissue in culture.

The method, referred to as the Volumetric Centrifugation Method (VCM), was published in the March 28, 2014 Nature Communications.

The new discovery will have a major impact on the hazard assessment of engineered nanoparticles, enabling risk assessors to perform accurate hazard rankings of nanomaterials using cellular systems. Furthermore, by measuring the composition of nanomaterial agglomerates in physiologic fluids, it will allow scientists to design more effective nano-based drug delivery systems for nanomedicine applications.

“The biggest challenge we have in assessing possible health effects associated with nano exposures is deciding when something is hazardous and when it is not, based on the dose level. At low levels, the risks are probably miniscule,” said senior author Philip Demokritou, associate professor of aerosol physics in the Department of Environmental Health at HSPH. “The question is: At what dose level does nano-exposure become problematic? The same question applies to nano-based drugs when we test their efficiency using cellular systems. How much of the administered nano-drug will come in contact with cells and tissue? This will determine the effective dose needed for a given cellular response,” Demokritou said.

Federal regulatory agencies do not require manufacturers to test engineered nanoparticles, if the original form of the bulk material has already been shown to be safe. However, there is evidence that some of these materials could be more harmful in the nanoscale — a scale at which materials may penetrate cells and bypass biological barriers more easily and exhibit unique physical, chemical, and biological properties compared to larger size particles. Nanotoxicologists are struggling to develop fast and cheap toxicological screening cellular assays to cope with the influx of vast forms of engineered nanomaterials and avoid laborious and expensive animal testing. However, this effort has been held back due to the lack of a simple-to-use, fast, method to measure the dose-response relationships and possible toxicological implications. While biological responses are fairly easy to measure, scientists are struggling to develop a fast method to assess the exact amount or dose of nanomaterials coming in contact with cells in biological media.

“Dosimetric considerations are too complicated to consider in nano-bio assessments, but too important to ignore,” Demokritou said. “Comparisons of biological responses to nano-exposures usually rely on guesstimates based on properties measured in the dry powder form (e.g., mass, surface area, and density), without taking into account particle-particle and particle-fluid interactions in biological media. When suspended in fluids, nanoparticles typically form agglomerates that include large amounts of the suspending fluid, and that therefore have effective densities much lower than that of dry material. This greatly influences the particle delivery to cells, and reduces the surface area available for interactions with cells,” said Glen DeLoid, research associate in the Department of Environmental Health, one of the two lead authors of the study. “The VCM method will help nanobiologists and regulators to resolve conflicting in vitro cellular toxicity data that have been reported in the literature for various nanomaterials. These disparities likely result from lack of or inaccurate dosimetric considerations in nano-bio interactions in a cellular screening system,” said Joel Cohen, doctoral student at HSPH and one of the two lead authors of the study.

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

Estimating the effective density of engineered nanomaterials for in vitro dosimetry by Glen DeLoid, Joel M. Cohen, Tom Darrah, Raymond Derk, Liying Rojanasakul, Georgios Pyrgiotakis, Wendel Wohlleben, & Philip Demokritou. Nature Communications 5, Article number: 3514 doi:10.1038/ncomms4514 Published 28 March 2014

This paper is behind a paywall but a free preview is available via ReadCube Access.

Ecotoxicology and environmental fate of manufactured nanomaterials—testing guidelines from Organization for Economic Cooperation and Development (OECD)

The Organization for Economic Cooperation and Development (OECD) has released guidelines for testing manufactured nanomaterials according to a March 11, 2014 news item on Nanowerk,

As part of its Programme on the Safety of Manufactured Nanomaterials, and in particular work on the testing and assessment of manufactured nanomaterials, OECD initiated a series of expert meetings to improve the applicability of the OECD Test Guidelines to nanomaterials. With this in mind, the Working Party on Manufactured Nanomaterials agreed to address the ecotoxicology and environmental fate of manufactured nanomaterials.

The OECD Expert Meeting on Ecotoxicology and Environmental Fate took place on 29th-31st January 2013 in Berlin, Federal Press Office. The event was hosted by the German delegation and funded by the German Federal Ministry of the Environment, Nature Conservation and Nuclear Safety (BMU) as well as the United States Environment Protection Agency (US EPA).

Three documents were published one of which being a preview,

The OECD expert meeting on ecotoxicology and environmental fate — Towards the development of improved OECD guidelines for the testing of nanomaterials by Dana Kühnel and Carmen Nickel. Science of The Total Environment Volume 472, 15 February 2014, Pages 347–353 http://dx.doi.org/10.1016/j.scitotenv.2013.11.055

This document is open access.

The report itself,

OECD. ENVIRONMENT DIRECTORATE.
JOINT MEETING OF THE CHEMICALS COMMITTEE AND
THE WORKING PARTY ON CHEMICALS, PESTICIDES AND BIOTECHNOLOGY. Environment, Health and Safety Publications
Series on the Safety of Manufactured Nanomaterials. ENV/JM/MONO(2014)1

ECOTOXICOLOGY AND ENVIRONMENTAL FATE OF MANUFACTURED NANOMATERIALS:
TEST GUIDELINES Expert Meeting Report
Series on the Safety of Manufactured Nanomaterials No. 40

Ecotoxicology and Environmental Fate of Manufactured Nanomaterials: Test Guidelines

There’s an addendum which includes the presentations made at the meeting (you can find both the report, proper, and the addendum on this page scroll to report no. 40),

OECD. ENVIRONMENT DIRECTORATE JOINT MEETING OF THE CHEMICALS COMMITTEE AND
THE WORKING PARTY ON CHEMICALS, PESTICIDES AND BIOTECHNOLOGY. Environment, Health and Safety Publications. ENV/JM/MONO(2014)1/ADD

ADDENDUM TO EXOTOXICOLOGY AND ENVIRONMENTAL FATE OF MANUFACTURED
NANOMATERIALS: TEST GUIDELINES

Series on the Safety of Manufactured Nanomaterials No. 40
Ecotoxicology and Environmental Fate of Manufactured Nanomaterials:
Test Guidelines.

As it can get a little tricky accessing OECD documents, I’ve tried to give a couple different links and as much identifying information as possible. Good luck!

Danish scientists provide insights into celllular response to silver nanoparticles

The conclusions are concerning but the scientists at the University of Southern Denmark are careful to note that this research on silver nanopartices was performed in a laboratory setting which does not necessarily predict what might happen under real life conditions.

As for the research itself, a Feb. 28, 2014 news item on Azonano has this to say,

Endocrine disrupters are not the only worrying chemicals that ordinary consumers are exposed to in everyday life. Also nanoparticles of silver, found in e.g. dietary supplements, cosmetics and food packaging, now worry scientists. A new study from the University of Southern Denmark shows that nano-silver can penetrate our cells and cause damage.

Silver has an antibacterial effect and therefore the food and cosmetic industry often coat their products with silver nanoparticles. Nano-silver can be found in e.g. drinking bottles, cosmetics, band aids, toothbrushes, running socks, refrigerators, washing machines and food packagings.

“Silver as a metal does not pose any danger, but when you break it down to nano-sizes, the particles become small enough to penetrate a cell wall. If nano-silver enters a human cell, it can cause changes in the cell”, explain Associate Professor Frank Kjeldsen and PhD Thiago Verano-Braga, Department of Biochemistry and Molecular Biology at the University of Southern Denmark.

A Feb. 27, 2014 University of Southern Denmark news release, which originated the news item, provides more detail about the research,

The researchers examined human intestinal cells, as they consider these to be most likely to come into contact with nano-silver, ingested with food.

“We can confirm that nano-silver leads to the formation of harmful, so called free radicals in cells. We can also see that there are changes in the form and amount of proteins. This worries us”, say Frank Kjeldsen and Thiago Verano-Braga.

A large number of serious diseases are characterized by the fact that there is an overproduction of free radicals in cells. This applies to cancer and neurological diseases such as Alzheimer’s and Parkinson’s.

Kjeldsen and Verano-Braga emphasizes that their research is conducted on human cells in a laboratory, not based on living people. They also point out that they do not know how large a dose of nano-silver, a person must be exposed to for the emergence of cellular changes.

“We don’t know how much is needed, so we cannot conclude that nano-silver can make you sick. But we can say that we must be very cautious and worried when we see an overproduction of free radicals in human cells”, they say.

Nano-silver is also sold as a dietary supplement, promising to have an antibacterial, anti-flu and cancer-inhibatory effect. The nano-silver should also help against low blood counts and bad skin. In the EU, the marketing of dietary supplements and foods with claims to have medical effects is not allowed. But the nano-silver is easy to find and buy online.

In the wake of the SDU-research, the Danish Veterinary and Food Administration now warns against taking dietary supplements with nano-silver.

“The recent research strongly suggests that it can be dangerous”, says Søren Langkilde from the Danish Veterinary and Food Administration to the Danish Broadcasting Corporation (DR).

The researchers supplied this image to illustrate the abstract for their paper (link and citation to follow),

Courtesy University of Southern Denmark

Courtesy University of Southern Denmark

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

Insights into the Cellular Response Triggered by Silver Nanoparticles Using Quantitative Proteomics by Thiago Verano-Braga, Rona Miethling-Graff, Katarzyna Wojdyla, Adelina Rogowska-Wrzesinska, Jonathan R. Brewer, Helmut Erdmann, and Frank Kjeldsen. ACS Nano, Article ASAP DOI: 10.1021/nn4050744 Publication Date (Web): February 10, 2014
Copyright © 2014 American Chemical Society

This paper is behind a paywall.

Silver ions in the environment

Earlier this week (Feb. 24, 2014), I published a post featuring Dr. Andrew Maynard, Director of the University of Michigan’s Risk Science Center in an introductory video describing seven surprising facts about silver nanoparticles. For those who want to delve more deeply, there’s a Feb. 25, 2014 news item on Nanowerk describing some Swiss research into silver nanoparticles and ions in aquatic environments,

It has long been known that, in the form of free ions, silver particles can be highly toxic to aquatic organisms. Yet to this day, there is a lack of detailed knowledge about the doses required to trigger a response and how the organisms deal with this kind of stress. To learn more about the cellular processes that occur in the cells, scientists from the Aquatic Research Institute, Eawag [Swiss Federal Institute of Aquatic Science and Technology], subjected algae to a range of silver concentrations.

In the past, silver mostly found its way into the environment in the vicinity of silver mines or via wastewater [emphasis mine] emanating from the photo industry. More recently, silver nanoparticles have become commonplace in many applications – as ingredients in cosmetics, food packaging, disinfectants, and functional clothing. Though a recent study conducted by the Swiss National Science Foundation revealed that the bulk of silver nanoparticles is retained in wastewater treatment plants, only little is known about the persistence and the impact of the residual nano-silver in the environment.

The Feb. 25, 2014 Eawag media release, which originated the news item, describes the research in further detail,

Smitha Pillai from the Eawag Department of Environmental Toxicology and her colleagues from EPF Lausanne and ETH Zürich studied the impact of various concentrations of waterborne silver ions on the cells of the green algae Chlamydomonas reinhardtii. Silver is chemically very similar to copper, an essential metal due to its importance in several enzymes. Because of that, silver can exploit the cells’ copper transport mechanisms and sneak into them undercover. This explains why, already after a short time, concentrations of silver in the intracellular fluid can reach up to one thousand times those in the surrounding environment.

A prompt response

Because silver damages key enzymes involved in energy metabolism, even low concentrations can cut photosynthesis and growth rates by a half in just 15 minutes. Over the same time period, the researchers also detected changes in the activity of about 1000 other genes and proteins, which they interpreted as a response to the stressor – an attempt to repair silver-induced damage. At low concentrations, the cells’ photosynthesis apparatus recovered within five hours, and recovery mechanisms were sufficient to deal with all but the highest concentrations tested.

A number of unanswered questions

At first glance, the results are reassuring because the silver concentrations that the algae are subject to in the environment are rarely as high as those applied in the lab, which allows them to recover quickly – at least externally. But the experiments also showed that even low silver concentrations have a significant effect on intracellular processes and that the algae divert their energy to repairing damage incurred. This can pose a problem when other stressors act in parallel, such as increased UV-radiation or other chemical compounds. Moreover, it remains unknown to this day whether the cells have an active mechanism to shuttle out the silver. Lacking such a mechanism, the silver could have adverse effects on higher organisms, given that algae are at the bottom of the food chain.

You can find the researchers’ paper here,

Linking toxicity and adaptive responses across the transcriptome, proteome, and phenotype of Chlamydomonas reinhardtii exposed to silver by Smitha Pillai, Renata Behra, Holger Nestler, Marc J.-F. Suter, Laura Sigg, and Kristin Schirmer. Proceedings of the National Academy of Sciences (PNAS) – early edition 18.February 2014, www.pnas.org/cgi/doi/10.1073/pnas.1319388111

The paper is available through the PNAS open access option.

I have published a number of pieces about aquatic enviornments and wastewater and nanotechnology-enabled products as useful for remediation efforts and as a source of pollution. Here’s a Feb. 28, 2013 posting where I contrasted two pieces of research on silver nanoparticles. The first was research in an aquatic environment and the other concerned wastewater.

Czech veterinary research institute tracks nanoparticles

When I first saw the Jan. 7, 2014 news item on Azonano, I was expecting to see some cute animal images mixed with the ‘nano’ talk. While there’s no mild amusement to be had, there is plenty of ‘nano’ talk concerning the work being done at the Veterinary Research Institute (Brno,  Czech republic) on characterizing nanoparticles using some new equipment (Note: Links have been removed),

Malvern [which owns the company, NanoSight] reports on how NanoSight’s Nanoparticle Tracking Analysis, NTA, is being applied in the Veterinary Research Institute, Brno, Czech Republic in the research group of Dr. Jaroslav Turanek in the Department of Pharmacology and Immunotherapy).

The central theme of Dr. Jaroslav Turanek’s research group (Department of Pharmacology and Immunotherapy) at the Veterinary Research Institute in Brno is to apply synthetic and bioorganic chemistry. This work is performed in collaboration with King’s College London and the Institute of Organic Chemistry and Biochemistry, Prague, for the design and construction of therapeutic nanoparticles to develop drug delivery systems (anticancer and antiviral drugs) and nanocarriers for construction of recombinant vaccines.

In parallel, the research group of Dr. Miroslav Machala (Department of Chemistry and Toxicology) at Veterinary Research Institute focuses upon environmental nanoparticulate pollutants. Characterization of airborne particles is conducted using electron microscopy, but in vitro tests on cell culture require knowledge of the real structure of nanoparticles in the tissue culture medium (e.g. aggregation). This enables the group to draw the correct conclusions from in vitro toxicological experiments which can be affected by differences in local nanoparticle concentration owing to sedimentation. Detailed particle distribution and kinetics of aggregation in this heterogeneous system is impossible to obtain using electron microscopy and hence Nanoparticle Tracking Analysis, NTA, is the method of choice. It is noted that some metastable aggregates can disaggregate due to high dilution of the sample required for NTA analysis. For this reason, Dynamic Light Scattering, DLS, and NTA are used as suitable complementary methods in the laboratory.

The Jan. 7, 2014 Malvern Instruments press release on biospace.com, which originated the news item, provides a quote from Dr. Jaroslav Turanek’ describing the NTA system,

Explaining their choice of NanoSight, Dr Turanek said “We chose NTA as a convenient and rapid method for characterization of nanoparticles in heterogeneous preparations like liposomes and their complexes with proteins, DNA and polysaccharides. A set of these techniques is used for the complex characterization of the structure of the nanoparticles, the kinetics of their preparation, the dynamics of morphological transformation and, finally, their stability. NTA perfectly fits our needs and has become a standard method in our methodological portfolio. The most advantageous feature of NTA is that it makes it possible to visualize each nanoparticle and then to obtain more detailed size distributions based on individual particle measurements. DLS is used as precise complementary method for the characterization of nanoparticles below 20 nm for proteins and other biopolymers. Combination of these two methods, NTA and DLS, with separation methods (GPC, FFF) and electron microscopy is preferred to get the full insight to structure and dynamics of nanoparticles in our sample systems.”

It took me quite a while to realize that nanoparticles in a sample are not necessarily homogenous, i.e., similar in size, etc. Unconsciously, I had applied my notions of manufacturing where items are made (stamped, poured into moulds, etc.)  to be identical. As far as I’m aware there is no such production process for nanoparticles which makes characterizing them an important task if the purpose is to better understand their properties.

You can find out more about Malvern Instruments here and about NanoSight here.

Human immune system and nanotoxicology in Québec (Canada)

At this point it’s starting to seem like there are thousands and thousands of nanotoxicology studies so the announcement of a new study based in Québec (Canada) didn’t immediately cause excitement  until I caught sight of the word ‘inflammation” which casts a newish light on the topic. From the Dec. 4, 2013 news item on Azonano,

… Professor Girard [Professor Denis Girard INRS–Institut Armand-Frappier Research Centre] will focus on the effects of NPs [nanoparticles] on human immune system cells (eosinophils) that play a key role in inflammation.

“Several studies on NPs have examined how tissues react in contact with these tiny foreign bodies,” said Girard. “Researchers have found that eosinophils flock to the contact site, but they have not examined the phenomenon in greater detail.” To further investigate why eosinophils come into contact with NPs and the role they play, protocols require expertise in both nanotoxicology and immunology, which is rare.

The Nov. 28, 2013 INRS [Institut national de la recherche scientifique] Université news release by Stéphanie Thibault, which originated the news item, delves into the issue of inflammatory responses,

According to Professor Girard, understanding the inflammatory response is currently a priority in the field of nanotoxicology. For a number of years, researchers have been observing links between exposure to NPs and asthmatic symptoms in some animals. Does the human body undergo similar inflammation upon contact with NPs? In the absence of any standards for workers, it’s best to take a closer look, insists Girard. “At this time, nanoparticles have not been properly identified and are often handled without protection. If they enter the body through the skin, respiratory tract, or even ingestion, we have no idea what happens next.” In his lab, a variety of approaches will help further understanding of how nanoparticles of different types and sizes interact. Cellular processes will be examined in detail.

 

At the rate at which NPs are being developed, Girard could be conducting systematic nanotoxicology studies for many years to come. “I will of course need the support of a strong team,” said Girard. “I already have one I am very proud of, and it will be expanded for the new project.” …

I gather there are going to be some jobs generated from this grant,

His research is being funded by Institut de recherche Robert-Sauvé en santé et en sécurité du travail (IRSST), which will award him a renewable $300,000 grant for the next three years.

… The IRSST grant will be used to hire staff and student researchers.

While I have heard of the IRSST before,, the INRS is new to me. Here’s more from the INRS English language homepage,

INRS (Institut national de la recherche scientifique) is one of Canada’s top universities in terms of research intensity (funding per faculty member). It brings together 150 professors, over 700 graduate and postgraduate students, and a hundred postdoctoral researchers at four centers in Montreal, Québec City, Laval, and Varennes. Conducting applied and fundamental research essential to the advancement of science in Quebec and around the world, our research teams plays a critical role in finding solutions to problems facing our society. Founded in 1969, INRS is one of the nine establishments that make up the Université du Québec network.

 

“The Institute is dedicated to fundamental and applied research, graduate studies, and the training of researchers. In keeping with its mission and objectives as a research university, the Institute specifically gears its activities towards Quebec’s economic, social, and cultural development, as well as the transfer of knowledge and technology stemming from all its fields of study.” INRS letters patent, 1999

There you have it.

2014 Internatonal NanoSafety Congress in Iran extends deadline for submissions to Dec. 15, 2013

A Nov. 11, 2013 news item on Nanowerk highlights the 2014 Iran International NanoSafety Congress and the deadline extension,

The deadline for paper submission to Iran International Nanosafety Congress was extended to 15 December 2013.
Iran Nanosafety Congress will be held in Tehran University of Medical Sciences in association with Iran Nanosafety Network (INSN) of Iran Nanotechnology Initiative Council on 19-20 February [2014] to guarantee the safe and continuous development of nanotechnology, give correct information about nanosafety, identify active bodies in the field of nanosaftey and develop cooperation with other countries.
The scope of the congress is as follows:
– Exposure assessment
– Methodology: characterization, detection, and monitoring
– Occupational and environmental interactions
– Toxicology
– Ecotoxicology and life cycle analysis
– Standardization and regulations

The homepage for the Iran International NanoSafety Congress provides more information,

Dear Colleagues,
On behalf of the scientific and executive committees, it is our great pleasure to cordially invite you to attend the Iran NanoSafety Congress 2014 (INSC 2014) which will be held at the Ghods Auditorium in Tehran University of Medical Sciences, on 19-20 February 2014.

This Congress is jointly organized by the Iran Nanosafety Network (INSN) of Iran Nanotechnology Initiative Council (INIC) and Tehran University of Medical Sciences (TUMS), supported by Iran Nanohealth Committee of Food & Drug Organization (INC), Iranian Environmental Mutagen Society (IrEMS) and Iranian Society of Nanomedicine (ISNM).  The “Iran NanoSafety Congress 2014″ aims to cover all safety aspects of nanomaterials in human and environment. This Congress is focused on novel approaches and technologies being used to properly assess the safety, toxicity, and risk of nanomaterial for occupational and environmental health. The scientific program will consist of keynote/distinguished lectures, symposia, workshops, discussion panels and poster sessions. This congress will provide attendees good opportunities to meet scientists from all over the world to exchange the ideas and to launch national and international collaborations in different aspects of  Nanosafety. The organizing committee is also planning a variety of unique social programs to provide the chance for participants to enjoy from fascinating Iranian culture and warm spirit of friendship.

We look forward to welcoming you and your active participation in the INSC 2014 in Tehran, I.R. Iran.

Good luck with getting your submission in on time.

Europe’s open access nanotoxicology database: old news?

An Oct. 7, 2013 news item on Nanowerk announces the launch of an open-access database for nanotoxicology materials,

The rise of potential health hazards has given rise to a new discipline — nanotoxicity — the study of toxicity as a result of nanomaterials. Work undertaken by the project ‘Nano health-environment commented database’ (NHECD) has culminated in a completely open-access database that incorporates a mechanism for updating the knowledge repository.

A major factor taken into account by NHECD was that the users come from many different groups including the press, research institutions and governmental bodies. Not only does the NHECD database hold unstructured data like scientific papers, it also allows for automatic updating. The database can thus hold a dynamic developing collection of published data on environmental and health effects after exposure to nanoparticles.

The NHECD database homepage offers more information about itself,

What is NHECD and what can be found here?

NHECD is a free access, robust and sustainable web based information system including a knowledge repository on the impact of nanoparticles on health, safety and the environment. It includes a robust content management system (CMS) as its backbone, to hold unstructured data (e.g., scientific papers and other relevant publications). …

 Discover Our Intelligent Search

Our intelligent search is a unique method to target the information you need. This search feature is especially crafted for the needs of researchers in the nanoscience field. This feature includes among other capabilities the power to search by model, experiment and nanoparticles attributes. …

I did try a couple of searches ‘silver nanoparticles’ and ‘carbon nanotubes’ to no avail.

This project puts me in mind of the GoodNanoGuide, which has somewhat similar aspirations,

The mission of the GoodNanoGuide is to provide an Internet-based collaboration platform specially designed to enhance the ability of experts to exchange ideas on how best to handle nanomaterials in an occupational setting. It is meant to be an interactive forum that fills the need for up-to-date information about current good practices for managing nanomaterials in a work-related environment, highlighting new practices as they develop.

The goal of the GoodNanoGuide is to create a central repository for good practices for safely handling nanomaterials that can be used and contributed to by people from all over the world.

In fact, I noted this similarity in a July 13, 2009 posting titled: Good Nano Guide and the UK’s NHECD project complementary? plus the Finnish, the Canadians, nanotechnology and innovation. I don’t understand why the NHECD is being publicized at this point in time as the website does not seem to be fully populated (blank webpages) and as I noted I had difficulty running a search.

Toxicity, nanoparticles, soil, and Europe’s NANO-ECOTOXICITY Project

I have featured pieces on nanoparticles, toxicity, and soil in the past (this Aug. 15, 2011 posting about Duke University’s mesocosm project is probably the most relevant) but this study is the first one I’ve seen focusing on earthworms. From the Sept. 23, 2013 news item on Nanowerk (Note: A link has been removed),

From the clothes and make-up we wear to the electronic devices we use every day, nanotechnology is becoming ubiquitous. But while industry has mastered the production of such materials, little is known about their fate once their service life comes to an end. The NANO-ECOTOXICITY project looked into their impact on soil organisms.

The Sept. 23, 2013 CORDIS (European Commission Community Research and Development Information Service) news release, which originated the new item, offers a Q&A (Question and Answer) with the project research leader,

Dr Maria Diez-Ortiz, research leader of the NANO-ECOTOXICITY project, tells us about her research findings and how she expects them to help increase knowledge and shape tools allowing for standard environmental hazard and risk-assessment methodologies.

What is the background of the NANO-ECOTOXICITY project?

Nanotechnology is based on the idea that, by engineering the size and shape of materials at the scale of atoms, i.e. nanometres (nm), distinct optical, electronic, or magnetic properties can be tuned to produce novel properties of commercial value. However, there is an obvious concern that such novel properties may also lead to novel behaviour when interacting with biological organisms, and thus to potentially novel toxic effects.

Since nanoparticles (NPs) are similar in size to viruses, their uptake by and transport through tissues are based on mechanisms distinct from those of molecular uptake and transport. Therefore, there is concern that standard toxicological tests may not be applicable or reliable in relation to NPs, hence compromising current risk-assessment procedures.

The majority of research on nano-safety in the environment has so far focused on the aquatic environment. Current research on environmental fate, however, indicates that soils will become the biggest environmental sink for nanoparticles. Following their entry into liquid waste streams, nanoparticles will pass through wastewater-treatment. processes, ending up in waste sludge which may accumulate in the agricultural land where this sludge is often applied.

What are the main objectives of the project?

This project deals with the toxicokinetics – that is, the rate at which a chemical enters a body and affects it – of metal nanoparticles coming into contact with soil-dwelling organisms. The aim is to determine NPs’ fate and effects in terrestrial ecosystems by means of case studies with zinc oxide and silver NPs, which represent different fate kinetics.

The project’s main objectives are to assess the toxicity of metal nanoparticles in soils in the short and long term; the main route of exposure for earthworms and whether it differs from those of ionic metals; and, finally, the influence of the exposure media on metal nanoparticle toxicity.

What is new or innovative about the project and the way it is addressing these issues?

We have been running a long-term study where soils with AgNP [silver nanoparticles] were stored and left to age for up to a year; their toxicity was tested at the start and after three, seven and 12 months of ageing. The results showed that silver toxicity increased over time, meaning that short-term standard toxicity tests may underestimate the environmental risk of silver nanoparticles.

In parallel, we found that organisms exposed to silver nanoparticles in short-term studies accumulated higher silver concentrations than organisms that were exposed to the same mass concentration of ionic silver. However, these NP exposed organisms actually suffered lower toxic effects. This observation contradicts the prevailing assumption in toxicology that the internalised concentration is directly related to chemical concentration at the target site and hence to its toxicity. This observation creates a new paradigm for nano-ecotoxicology.

What is not yet known is whether the accumulated NP metal may in the longer-term ultimately become toxic (e.g. through dissolution and ion release) in cells and tissues where AgNPs may be stored. Should this occur, the high concentrations accumulated may ultimately result in greater long-term toxicity for NPs than for ionic forms. This may reveal these accumulated NPs as internalised ‘time bombs’ relevant to long-term effects and toxicity.

However, it has to be borne in mind that the redicted environmental concentrations resulting from current use of nanoparticles (e.g. results from EU projects like NANOFATE2) are many times smaller than those used in these studies, meaning that such accumulations of nanoparticle-related silver are unlikely to occur in the environment or, ultimately, in humans.

What difficulties did you encounter and how did you solve them?

The main problems encountered relate to the tracking of nanoparticles inside the tissues and soils, as both are complex matrices. The analysis of the particles is a challenge in itself, even when in water, but to get information about their state in these matrices often requires unrealistic exposure concentrations (due to low detection limits of the highly specialised techniques used for analysis) or extraction of the particles from the matrices, which could potentially change the state of the particles.

In this project, I travelled to University of Kentucky to work with Jason Unrine and used gentle water-based extractions of soil samples immediately before analysing them using ‘Field-flow fractionation’ and ‘Inductively coupled plasma mass spectrometry’ to identify the state of nanoparticles in my aged soils.

To look at what form (speciation) of silver and zinc from the nanoparticle exposures could be found inside worms I collaborated with NANOFATE researchers at Cardiff University who fixed and thinly sectioned the worm tissues. I was lucky to be given the time to use specialist facilities like the UK’s Diamond Light Source synchrotron to investigate where and in what form the metals and potential nanoparticles could be found in these tissues.

The main challenge is that as soon as you take nanoparticles out of the manufacturers’ bottle they start changing, particularly when put into environments likes natural soils and waters, or even organisms. Therefore a lot of characterisation is needed during exposure to establish the state of the nanoparticles the organisms have been exposed to and how fast they are changing from pristine particles to dissolved ions, or particles with completely different surfaces.

Technical solutions to characterisation have been found during this short project, but this will remain a logistical challenge for many years to come as the analysis equipment is still very specialised and expensive and therefore not generally available.

What are the concrete results from the research so far?

The project has helped us draw various conclusions regarding the impact of NPs on the environment and how to assess them. First, we now know that soil acidity, or pH, influences the dissolution and toxicity of ZnO nanoparticles [zinc oxide].

Then, we found that toxicity of silver nanoparticles’ increases over time and that the particles’ coating affects their toxicity to soil invertebrates.

As previously mentioned, earthworms exposed to silver nanoparticles for 28 days accumulated higher silver concentrations than earthworms exposed to silver ions, without the excess silver from the nanoparticles having a toxic effect. [emphasis mine] Moreover, soil ingestion was identified as the main route of exposure to AgNP and ZnONP in earthworms.

How can industry and decision-makers ensure that nanomaterials do not impact our environment?

We hope that this project, and the larger EU project NANOFATE to which it is linked, will provide knowledge and tools enabling standard environmental-hazard and risk-assessment methodologies to be applied to engineered nanoparticles (ENPs) with just a few judicious modifications. The current systems and protocols for chemical risk assessment have been developed over decades, and where no novel toxic mechanisms exist, our results tend to say that nano fits in as long as we measure the right things and characterise realistic exposures properly.

Our research aims to determine the minimum methodological tweaks needed. So far everything indicates that the potential benefits from nanotechnology can be realised and managed safely alongside other chemicals. While we are fairly confident at this stage that ENPs impose no greater acute effects on important biological parameters – like reproduction – than their ionic forms, the NANO-ECOTOXICITY results demonstrate that we have some way to go before we can state loud and clear that we do not believe there is any novel low-level or long-term effect.

As for all chemicals, proving such a negative is impossible using short-term tests. We think the final conclusions by industry and regulators on safe use of nanoparticles should and will have to be made according to a ‘weight of evidence’ approach – proving there is a gap between predicted likely exposure levels and those levels seen to cause any effects or accumulations within ecosystem species.

What are the next topics for your research?

This project has finished but the next step for any other funding opportunity would be to address increasingly environmentally relevant exposure scenarios by analysing how nanoparticles modify in the environment and interact with living tissues and organisms at different trophic levels. I would like to investigate nanoparticle transformation and interactions in living tissues. To date, the studies that have identified this ‘excess’ accumulation of non-toxic metal loads in nanoparticleexposed organisms have only been short term.

Apart from the obviously increased food-chain transfer potential, is also not known whether, over the longer term, the accumulated NP-derived metal ultimately becomes toxic when present in tissues and cells. Such transformation and release of metal ions within tissues may ultimately result in greater longterm toxicity for NPs than for ionic forms.

Furthermore, I want to test exposures in a functioning model ecosystem including interspecific interactions and trophic transfer. Since interactions between biota and nanoparticles are relevant in natural soil systems, caution is needed when attempting to predict the ecological consequences of nanoparticles based on laboratory assays conducted with only a single species. In the presence of the full complement of biological components of soil systems, complex NPs may follow a range of pathways in which coatings may be removed and replaced with exudate materials. Studies to quantify the nature of these interactions are therefore needed to identify the fate, bioavailability and toxicity of realistic ‘non-pristine’ forms of NPs present in real soil environments.

New to me was the material about ageing silver nanoparticles and their increased toxicity over time. While this is an interesting piece of information it’s not necessarily all that useful. It seems even with their increased uptake compared to silver ions, silver nanoparticles (Diez-Ortiz doesn’t indicate whether or not * they tested variously aged silver nanoparticles) did not have toxic effects on the earthworms tested.

The NANO-ECOTOXICITY website doesn’t appear to exist anymore but you can find the NANOFATE (Nanoparticle Fate Assessment and Toxicity in the Environment) website here.

* ‘not’ removed to clarify meaning, Oct. 9, 2013. (Note: I had on Oct. 8, 2013 removed ‘not’ in a second place from the sentence in an attempt t o clarify the meaning and ended up not making any sense at all.) Please read Maria Diez-Ortiz in the Comments, as she clarifies matters in a way I could never hope to.

Nanomaterials, toxicology, and alternatives to animal testing

It seems that alternatives to animal testing may offer some additional capabilities for nanotoxicology studies according to an Aug. 21, 2013 news item on Nanowerk,

A group of international experts from government, industry and academia have concluded that alternative testing strategies (ATSs) that don’t rely on animals will be needed to cope with the wave of new nanomaterials emerging from the boom in nanoscience and nanotechnology. …

… Tests on laboratory mice, rats and other animals have been the standard way of checking new materials for health and environmental effects. Since those tests are costly, labor-intensive and time-consuming, workshop participants considered whether ATSs could have a larger role in checking the safety of ENMs [engineered nanomaterials].

They concluded that rapid cellular screening, computer modeling and other ATSs could serve as quick, cost-effective and reliable approaches for gathering certain types of information about the health and environmental effects of ENMs. “After lively discussions, a short list of generally shared viewpoints on this topic was generated, including a general view that ATS approaches for ENMs can significantly benefit chemical safety analysis,” they say.

The experts have had their consensus statement from the workshop published and before offering a citation for and a link to the statement, here’s the Abstract,

There has been a conceptual shift in toxicological studies from describing what happens to explaining how the adverse outcome occurs, thereby enabling a deeper and improved understanding of how biomolecular and mechanistic profiling can inform hazard identification and improve risk assessment. Compared to traditional toxicology methods, which have a heavy reliance on animals, new approaches to generate toxicological data are becoming available for the safety assessment of chemicals, including high-throughput and high-content screening (HTS, HCS). With the emergence of nanotechnology, the exponential increase in the total number of engineered nanomaterials (ENMs) in research, development, and commercialization requires a robust scientific approach to screen ENM safety in humans and the environment rapidly and efficiently. Spurred by the developments in chemical testing, a promising new toxicological paradigm for ENMs is to use alternative test strategies (ATS), which reduce reliance on animal testing through the use of in vitro and in silico methods such as HTS, HCS, and computational modeling. Furthermore, this allows for the comparative analysis of large numbers of ENMs simultaneously and for hazard assessment at various stages of the product development process and overall life cycle. [emphasis mine] Using carbon nanotubes as a case study, a workshop bringing together national and international leaders from government, industry, and academia was convened at the University of California, Los Angeles, to discuss the utility of ATS for decision-making analyses of ENMs. …

It seems that ATS has opened the door to more comprehensive testing (as per life cycles) than has previously been possible.

For the curious, here’s the citation for and the link to the published paper,

A Multi-Stakeholder Perspective on the Use of Alternative Test Strategies for Nanomaterial Safety Assessment by Andre E. Nel, Elina Nasser, Hilary Godwin, David Avery, Tina Bahadori, Lynn Bergeson #, Elizabeth Beryt, James C. Bonner, Darrell Boverhof, Janet Carter, Vince Castranova, J. R. DeShazo, Saber M. Hussain ●, Agnes B. Kane, Frederick Klaessig, Eileen Kuempel, Mark Lafranconi, Robert Landsiedel, Timothy Malloy, Mary Beth Miller, Jeffery Morris, Kenneth Moss, Gunter Oberdorster, Kent Pinkerton, Richard C. Pleus, Jo Anne Shatkin, Russell Thomas, Thabet Tolaymat, Amy Wang, and Jeffrey Wong. ACS Nano, Article ASAP DOI: 10.1021/nn4037927 Publication Date (Web): August 7, 2013

Copyright © 2013 American Chemical Society

This article is behind a paywall.