From in vivo to in vitro: The medical device testing paradigm shift.

Amid growing efforts to advance the replacement, reduction, and refinement of the use of animals in research, there is a growing recognition that in vitro testing of medical devices can be more effective, both in terms of cost and time, and also more reliable than in vivo testing. Although the technological landscape has evolved rapidly in support of these concepts, regulatory acceptance of alternative testing methods has not kept pace. Despite the acceptance by regulators of some in vitro tests (cytotoxicity, gene toxicity, and some hemocompatibility assays), many toxicity tests still rely on animals (irritation, sensitization, acute toxicity, reproductive/developmental toxicity), even where other industrial sectors have already abandoned them. Bringing about change will require a paradigm shift in current approaches to testing - and a concerted effort to generate better data on risks to human health from exposure to leachable chemicals from medical devices, and to boost confidence in the use of alternative methods to test devices. To help advance these ideas, stir debate about best practices, and coalesce around a roadmap forward, the JHU-Center for Alternatives to Animal Testing (CAAT) hosted a symposium believed to be the first gathering dedicated to the topic of in vitro testing of medical devices. Industry representatives, academics, and regulators in attendance presented evidence to support the unique strengths and challenges associated with the approaches currently in use as well as new methods under development, and drew next steps to push the field forward from their presentations and discussion.

Farm Animal Welfare and Human Health.

The paper examines the relationship between farm animal welfare, industrial farm animal production, and human health consequences. The data suggest that when the animal welfare of land-based farm animals is compromised, there are resulting significant negative human health consequences due to environmental degradation, the use of non-therapeutic levels of antibiotics for growth promotion, and the consequences of intensification. This paper accepts that even if meat and fish consumption is reduced, meat and fish will be part of the diet of the future. Industrial production modified from the current intensified systems will still be required to feed the world in 2050 and beyond. This paper identifies the concept of sustainable intensification and suggests that if farm animal welfare is improved, many of the human health consequences of intensified industrial production can be eliminated or reduced. In water-based farm animal production, many new systems are resulting in a product that actually protects the environment and can be done at industrial levels without the use of antibiotics.

The use of biomarkers of toxicity for integrating in vitro hazard estimates into risk assessment for humans.

The role that in vitro systems can play in toxicological risk assessment is determined by the appropriateness of the chosen methods, with respect to the way in which in vitro data can be extrapolated to the in vivo situation. This report presents the results of a workshop aimed at better defining the use of in vitro-derived biomarkers of toxicity (BoT) and determining the place these data can have in human risk assessment. As a result, a conceptual framework is presented for the incorporation of in vitro-derived toxicity data into the risk assessment process. The selection of BoT takes into account that they need to distinguish adverse and adaptive changes in cells. The framework defines the place of in vitro systems in the context of data on exposure, structural and physico-chemical properties, and toxicodynamic and biokinetic modeling. It outlines the determination of a proper point-of-departure (PoD) for in vitro-in vivo extrapolation, allowing implementation in risk assessment procedures. A BoT will need to take into account both the dynamics and the kinetics of the compound in the in vitro systems. For the implementation of the proposed framework it will be necessary to collect and collate data from existing literature and new in vitro test systems, as well as to categorize biomarkers of toxicity and their relation to pathways-of-toxicity. Moreover, data selection and integration need to be driven by their usefulness in a quantitative in vitro-in vivo extrapolation (QIVIVE).

The role of an academic centre.

On FRAME's 40th anniversary, I had the opportunity to examine FRAME and CAAT's missions as closely linked to those of their universities. The roles of education, research and service are key, both to the universities and to our two centres. By examining the current programmes, and identifying the needs of the future, the research activities, policy studies and training, it becomes clear that the Three Rs of alternatives contribute significantly to our respective universities' missions.

Meeting report: hazard assessment for nanoparticles--report from an interdisciplinary workshop.

In this report we present the findings from a nanotoxicology workshop held 6-7 April 2006 at the Woodrow Wilson International Center for Scholars in Washington, DC. Over 2 days, 26 scientists from government, academia, industry, and nonprofit organizations addressed two specific questions: what information is needed to understand the human health impact of engineered nanoparticles and how is this information best obtained? To assess hazards of nanoparticles in the near-term, most participants noted the need to use existing in vivo toxicologic tests because of their greater familiarity and interpretability. For all types of toxicology tests, the best measures of nanoparticle dose need to be determined. Most participants agreed that a standard set of nanoparticles should be validated by laboratories worldwide and made available for benchmarking tests of other newly created nanoparticles. The group concluded that a battery of tests should be developed to uncover particularly hazardous properties. Given the large number of diverse materials, most participants favored a tiered approach. Over the long term, research aimed at developing a mechanistic understanding of the numerous characteristics that influence nanoparticle toxicity was deemed essential. Predicting the potential toxicity of emerging nanoparticles will require hypothesis-driven research that elucidates how physicochemical parameters influence toxic effects on biological systems. Research needs should be determined in the context of the current availability of testing methods for nanoscale particles. Finally, the group identified general policy and strategic opportunities to accelerate the development and implementation of testing protocols and ensure that the information generated is translated effectively for all stakeholders.

Workgroup report: incorporating in vitro alternative methods for developmental neurotoxicity into international hazard and risk assessment strategies.

This is the report of the first workshop on Incorporating In Vitro Alternative Methods for Developmental Neurotoxicity (DNT) Testing into International Hazard and Risk Assessment Strategies, held in Ispra, Italy, on 19-21 April 2005. The workshop was hosted by the European Centre for the Validation of Alternative Methods (ECVAM) and jointly organized by ECVAM, the European Chemical Industry Council, and the Johns Hopkins University Center for Alternatives to Animal Testing. The primary aim of the workshop was to identify and catalog potential methods that could be used to assess how data from in vitro alternative methods could help to predict and identify DNT hazards. Working groups focused on two different aspects: a) details on the science available in the field of DNT, including discussions on the models available to capture the critical DNT mechanisms and processes, and b) policy and strategy aspects to assess the integration of alternative methods in a regulatory framework. This report summarizes these discussions and details the recommendations and priorities for future work.

In vitro and other alternative approaches to developmental neurotoxicity testing (DNT).

To address the growing need for scientifically valid and humane alternatives to developmental neurotoxicity testing (DNT), we propose that basic research scientists in developmental neurobiology be brought together with mechanistic toxicologists and policy analysts to develop the science and policy for DNT alternatives that are based on evolutionarily conserved mechanisms of neurodevelopment. In this article we briefly review in vitro and other alternative models and present our rationale for proposing that resources be focused on adapting alternative simple organism systems for DNT. We recognize that alternatives to DNT will not completely replace a DNT paradigm that involves in vivo testing in mammals. However, we believe that alternatives will be of great value in prioritizing chemicals and in identifying mechanisms of developmental neurotoxicity, which in turn will be useful in refining and reducing in vivo mammalian tests for exposures most likely to be hazardous to the developing human nervous system.

TestSmart and toxic ignorance.

There are a number of national and international efforts designed to screen chemicals for toxicity. Although the emphasis in terms of the specific chemicals is different, e.g. endocrine disruptors, children's health, High Production Volume (HPV), the European Registration, Evaluation and Authorisation of Chemicals (REACH) programme, the purpose is the same. Each is intended to evaluate the potential toxicity of chemicals to humans and, in some cases, to the environment. How best can these tasks be accomplished? The first need is to provide a realistic prioritisation of which chemicals need to be evaluated. Once identified, a defined decision-tree approach with an emphasis on short-term in vitro assays and new genomic technologies offers the greatest promise. The more practical matter of screening the chemicals would be by using a tiered decision-tree approach. Common features of the approach would be the use of three tiers. The first tier would be a screening/prioritisation tier, the second would provide an initial characterisation of toxicity, and the third would discern mode of action/biological activity. The intent of this approach is to provide a concept that will allow decisions to be made as to which chemicals need to be tested, provide some idea as to their toxicity and finally mode of action, and at the same time, taking into account the Three Rs, reduction, refinement and replacement. Thus, specific batteries of tests are not discussed, as these would need to be tailored to the specific chemicals of concern, e.g. endocrine disruptors, HPV. Neither are regulatory requirements factored into the concept, but the data that would be gathered should consider the possibility of eventual submission of the data obtained by in vitro and other non-traditional approaches by regulatory authorities.

Animals and alternatives: societal expectations and scientific need.

As Russell and Burch suggested more than 40 years ago, the most humane science is the best science. The path ahead is clear: pain and distress must be eliminated in animal experiments or reduced to an absolute minimum, and, as scientists, we must use the most humane approaches in our research. To accomplish the best science, we must train those who come after us in the principles and practice of humane science.

Use of animals in research: a science--society controversy? The American perspective: animal welfare issues.

My paper will focus on those events happening within the United States during the last year. The issue of including or excluding rats, birds and mice from inclusion under the Animal Welfare Act has been a difficult battle for both those that wish to exclude them and those that wish to include these animals under this legislation. As of the writing of this abstract, the Senate, which originally intended to include rats, birds and mice under the Animal Welfare Act, has passed an amendment which will permanently exclude their listing under this Act. During the last several years it has become clear that refinement, as one of the 3Rs, has and will become the most important set of activities to add humanness to animal experimentation. It is clear that refinement approaches provide the opportunity to possibly eliminate or significantly minimize any pain or distress in animal protocol. My presentation will focus on CAAT's (http://caat.jhsph.edu) activities in this important area. Understanding potential health hazards to environmental industrial chemicals has become a major focus of activity both in the US, Europe and Japan. These programs offer the first opportunity to provide information, in the public domain, on these chemicals. One of the consequences, however, is the potential requirement for large numbers of animals. In the presentation, I will focus on two approaches to significantly including the 3Rs in these important programs. Although it is common practice in Japan to recognize contributions of laboratory animals through a day of memorialization, this has not been the case in the United States. During the last year, several activities have been initiated to begin to institutionalize memorial services for animals used in research. As the host institution of Altweb (http://altweb.jhsph.edu), the alternative web site internationally, current statistics and accomplishments will be provided on its worldwide utilization.