Nanomaterial safety: An international collaboration

on in vitro testing strategies

(Nanowerk Spotlight) As a society, we are increasingly reliant on
high technology products that are made with rare and expensive
materials. They require high energy and material inputs, many of
which are potentially harmful to workers and end users and
problematic to manage responsibly across the life-cycle. The current
path of global consumerism is not sustainable. While there is no
internationally agreed definition, nanomaterials are substances that
fall in the size range of 1 to 100 nanometers (one billionth of one
meter) in at least one dimension, and by virtue of the small size they
may behave differently in the body or in the environment as
compared to macro-scale counterparts of the same
material.Engineered nanoscale materials (ENM or 'nanomaterials')
offer the potential to create safer and more effective products
through the use of smaller quantities of improved performance
materials. Currently nanomaterials are used to improve the
performance of life-saving drugs and medical technologies, to make
renewable energy more efficient, to make value added products from
industrial waste streams, to improve food, packaging, to lightweight
materials used in transportation systems, and to improve many of the
personal care products that we use every day. Nanomaterial
manufacture and use is expected to increase over the coming years
and despite the widespread use of nanomaterials in a variety of
consumer products, we are only beginning to understand the impacts
of these emerging materials on our health and the environment. To
this end, the University of Albertas Ingenuity Lab is collaborating
with the Society for Risk Analysis to evaluate the potential to use
alternative test strategies (ATS) to improve our ability to assess
nanomaterial toxicity and environmental impact.


Standard in vivo toxicology test methods that depend heavily
on the use of animals have long been used to assess chemical
safety. Existing and novel in vitro and in silico test methods
provide important alternatives to in vivo animal testing for
chemicals and potentially for ENM. Genotoxicity tests, for
example, are used to assess the mutagenic potential of
chemicals or nanomaterials in the replication of DNA in cells.
Driven in part by increasing market and regulatory
requirements for safer and more sustainable products, large
international infrastructure has developed for creating, testing
and validating in vitro test methods, and its use is expanding to
chemical and nanomaterial assessment (NSF, 2007). The goals
of reducing, refining and replacing animal testing (the
commonly cited 'three Rs') resonate with key and diverse
stakeholders including animal rights groups, the bioethics
community, the pharmaceutical industry, regulatory agencies
and the broader public.

Despite nearly a decade of effort in the conduct toxicology and
exposure research to inform the assessment of health and
environmental risks of nanomaterials, major gaps remain in the
ability to understand and quantify risks. While there is now a
large body of published data on carbon nanotubes and metal
oxide nanoparticles, concern has been raised that speculation
about nanomaterial risk has hardened into an assumption that
there are 'as-yet-to-be-discovered risks' that we must identify
and manage (Maynard, 2014) that demands extensive testing.

To ensure the appropriate management of risk, there is a need to
develop standardized, reliable approaches to evaluate and
classify novel substances through screening techniques and the
use of test methods that can reliably inform decision makers
about important priorities for additional testing that may be
required (Oberdorster et al, 2005; Nel et al, 2013). ATS
encompasses a variety of approaches that could address this
need.


Lori Sheremeta, the Assistant Director of Ingenuity Lab in
Edmonton Alberta and past Chair of the Society for Risk Analysis
(SRA) Emerging Nanoscale Materials Specialty Group (ENMSG), is
collaborating with U.S.-based nanomaterials risk expert Jo Anne
Shatkin (an SRA Councilor and co-founder of the SRA ENMSG),
Environment Canada, Health Canada, the SRA ENMSG and others
on a Pilot Project with the Organization for Economic Co-operation
and Development (OECD) Working Party on Manufactured
Nanomaterials (WPMN) to develop a report on the State of the
Science for ATS for nanomaterials, catalogue of existing and
emerging ATS methods in a database; and develop a case study to
inform workshop deliberations and expert recommendations.

ATS approaches are regarded by many to have the potential for
rapid screening of large numbers and types of materials. They can
include a breadth of techniques including high throughput
screening methods (HTS), high content screening, computational
approaches, toxicogenomics, cell-based methods, in vitro assays
and non-mammalian whole animal models. The emergence of ATS
raises questions about how the results of these methods may be
used for assessing the potential risks of ENM. For instance, ATS
could be used in combination in a multiple models approach to
evaluate new ENM in a number of rapid assays and compare with
well-studied substances using in vivo testing; thereby identifying
ENM for additional testing in a more strategic fashion than is
possible through conventional testing approaches.

Predictive models are developed by categorizing toxicological data
through a concept of relational characteristics, with representative
substances in a category used to infer toxicological information
about other substances that can be assigned to the same category.
The development of predictive models, particularly computational
models, requires large amounts of standardized high quality data.
Critical to their acceptance, ATS must have advantages over the
current or traditional methods, including reliability in producing
consistent results, and 'predictiveness' in correctly identifying
potential for biological effects that would occur in humans or other
species of interest.


The use of models to assess the toxicity of substances has a long
history that developed over time to inform understanding of
chemical effects. Mammalian studies represent earlier models that
informed toxicity testing. As the number of chemicals in commerce
grew, alternative methods were sought that could inform decision
making more quickly and with less expense. With the advent of cell
culturing methods, new models emerged. Over time, these
approaches have grown to inform both human and ecological
toxicity testing. Predictive models have emerged from a better
understanding of the relationships between chemical properties
and behavior in the body or the environment. In vitro and
predictive methods represent alternatives to the more traditional
toxicology testing involving studies in mammalian organisms. These
methods continue to grow in complexity, and are frequently
applied as part of a testing strategy.

There are many international efforts to develop, as well as to
validate and standardize, these methods for chemicals, including
organizations such as the US National Toxicology Program
Interagency Center for the Evaluation of Alternative Toxicological
Methods (ICCVAM), the European Union Reference Laboratory
European Centre for the Validation of Alternative Methods (EURL
ECVAM), the Japanese Center for the Validation of Alternative
Methods (JacVAM), the Korean Centre for the Validation of
Alternative Methods (KoCVAM) and the OECD. There is wide
recognition that the diversity of NMs renders it impractical to use
traditional animal testing to evaluate safety, hence there is
significant interest in assessing the performance of both existing
and emerging alternative testing strategies for NMs. Further, the
EU directive REACH (Directive 2006/121/EC) requires replacing in
vivo testing, and there is widespread popular agreement about the
desire to limit animal testing. Finally, there is a need for more
biologically informative toxicology methods (Hartung, 2010;
Silbergeld et al, 2011; Landsiedel et al, 2009).

The Society for Risk Analysis (SRA), through the Emerging
Nanoscale Materials Specialty Group, is organizing a public
workshop on September 15-16, 2014 in Washington, D.C. to
investigate the use of ATS in risk analysis for nanoscale materials.
The workshop will convene a diverse group of international experts
and stakeholders to discuss how existing and novel in vitro assays
may be applied in a 'multiple models' approach to inform the risk
assessment of novel nanoscale materials in assessing hazard,
potency and exposure potential. This effort builds on past SRA
efforts and other recent expert meetings regarding the development
and use of HTS by examining the availability and applicability of
existing and novel ATS methods for a multiple models approach to
toxicity, environmental and exposure analysis of nanomaterials in
the risk analysis paradigm (Shatkin et al, 2010). The SRA Nano
Risk Analysis II workshop seeks to elaborate shared strengths and
gaps in support of a weight of evidence approach relying on ATS to
inform context specific decisions about risk from exposure to
novel nanoscale materials. The goal is to inform the development of
guidance for ATS use in overall approaches to those decisions.The
main objectives of the workshop are to:
assess the state of the science on HTS and ATS from a 'multiple
models' perspective to identify areas of common findings from
differing approaches, areas of greatest uncertainty, and priorities for
follow up in applied research toward risk assessment of ENM;
evaluate the ability to use data from ATS/HTS methods for
screening purposes combining suites of assays and comparing
well-studied substances to novel ones;
assess the ability to use a suite of ATS methods to amplify the
Weight of Evidence;
characterize uncertainty associated with predictive relationships
and propose strategies to address uncertainties;
elicit the perspectives of diverse stakeholders about the use of
HTS/ATS for screening purposes in risk analysis of ENM; and
develop a set of recommendations for these alternative approaches
to become more widely adopted for environmental, health and safety
decision making about ENM across the product life cycle. The
output of the workshop holds potential for transformation through
risk screening approaches that promote safer and more sustainable
material and technology development.This workshop will facilitate
collaboration and communication between experts working on ATS
and those who will use the resulting data for risk analysis and risk
management decisions. As a non-governmental organization, the
Society for Risk Analysis is positioned to bring the diverse
researchers, nanotechnology experts, users and concerned parties
together to identify how to move forward on the critical need to
conduct screening of novel and emerging nanoscale materials to
overcome the current 'case-by-case' evaluation approach that has
proven too intensive, contentious, and a barrier to commercialization
of safer substances. The published research, education, and policy
recommendations that result from this meeting, as well as the
interpersonal networks established are expected to substantially
influence the international nanotechnology research enterprise and
emerging public policy for more sustainable management of NMs
for the next decade.In the United States, the U.S. ToxCast program
has, as part of their 21st century toxicity screening program (NRC,
2007), tested 29 NMs with 62 in vitro test methods (Wang et al.
2013). Many researchers, including several from the University of
Alberta, have proposed and developed ATS to include a variety of
methods, some which are standardized for chemicals, and others
which take advantage of developments including advanced
biological mechanistic understanding, genomics, metabolomics,
automation and informatics. However, these existing as well as
emerging ATS have a short history with nanomaterials, and have not
yet proven to be reliable for quantitative estimation of ENM risk.
Still, several international efforts have developed ATS that have
potential to be used for screening purposes, and to guide further
testing priorities for regulatory decision making. The goal of the
September workshop by the Society for Risk Analysis is to explore
ways in which distinct ATS may be used for screening and
prioritizing the need for more extensive testing of novel ENM.
NANO RISK ANALYSIS (II)
A Workshop to Explore How a Multiple Models Approach can
Advance Risk Analysis of Nanoscale Materials
September 15-16, 2014
George Washington University, Washington, DC.
This workshop brings together experts from diverse disciplines to
explore ways in which Alternative Testing Strategies (ATS) may be
combined to create a Weight of Evidence (WOE) or multiple
models approach to inform context-specific decisions about risk
from exposure to novel nanoscale materials. The goal is to advance
a common understanding of the state of the science, early lessons,
current opportunities, and next steps for developing ATS for use in
decision making for nanoscale materials.
The SRA ENMSG is coordinating the workshop, with numerous co-
sponsoring organizations, including Alberta Ingenuity Lab, PETA
International Science Consortium, Ltd., American Chemistry
Council, Bergeson and Campbell, Keller and Heckman, along with
the Society for Toxicology, Society for Environmental Toxicology
and Chemistry and others. Workshop details are available online
at www.srananoworkshop.org, and will be updated there regularly.
If your organization is interested in becoming a co-sponsor for the
Nano Risk Analysis (II) workshop, or if you are interested in getting
involved in planning and preparation, please contact Jo Anne
Shatkin.
The 2014 workshop will bring together key experts and
organizations working on ATS and HTS together with those who
may rely on such data to evaluate the potential for their use in risk
analysis of engineered nanomaterials, specifically to address the
issue of risk screening using ATS to infer toxicological and
exposure characteristics of new materials based on their
comparative performance to well-studied substances. Participants
will include active risk analysis experts from SRA, representatives
from OECD WPMN, major international nanotechnology grantees in
academic research institutes and others working on or interested in
using novel screening methods for nanomaterials and comparative
methods to screen close analogues at the nanoscale. International
governmental and non-governmental and civil society
representatives involved in toxicology, risk assessment and
alternative methods advancement are being invited. Diverse
environmental, consumer, labor and animal rights groups, as well
as other professional societies and their members are invited to
participate. Industry associations and representatives have been
invited as well, as key developers and users of nanomaterials. The
rate of nanomaterial research and development coupled with their
use in commercial products make reliable and acceptable screening
and predictive approaches for nanomaterials imperative. Yet, for
nanomaterials, many emerging methods and strategies are not yet
validated, standardized or widely adopted.