Framework to Evaluate Exposure Relevance and Data Needs for Risk Assessment of Nanomaterials using in Vitro Testing Strategies.

This article presents a multistage framework for evaluating the strength of evidence of nanomaterial (NM) exposure characterization data to optimize the utility of in vitro testing strategies for human health risk assessment. This framework is intended to aid risk assessors in evaluating the relevance of data from in vitro tests and to optimize the development of new in vitro testing strategies. The initial stage frames the exposure scenarios of interest in advance of testing to incorporate aspects such as release points, route of exposure, biological and environmental transformations, dose metrics, and biological targets in subsequent stages. The second stage considers characterization in the context of a realistic exposure and the third stage involves designing a testing strategy based on expected exposure conditions. For the fourth and final stage, we propose a matrix approach to evaluate the strength of evidence obtained in the first three stages as a basis for determining the best combination of test conditions and analytical methods available to characterize and measure exposure based on the NM type. This approach can also be used to evaluate existing data for their relevance to the expected exposure scenario and to further develop and optimize in vitro testing strategies. Implementation of the proposed strategy will generate meaningful information on NM properties and their interaction with biological systems, based on realistic exposure scenarios, which will be cost effective and can be applied for assessing risk and making intelligent regulatory decisions regarding the use and disposal of NMs.

Mammalian gastrointestinal tract parameters modulating the integrity, surface properties, and absorption of food-relevant nanomaterials.

Many natural chemicals in food are in the nanometer size range, and the selective uptake of nutrients with nanoscale dimensions by the gastrointestinal (GI) tract is a normal physiological process. Novel engineered nanomaterials (NMs) can bring various benefits to food, e.g., enhancing nutrition. Assessing potential risks requires an understanding of the stability of these entities in the GI lumen, and an understanding of whether or not they can be absorbed and thus become systemically available. Data are emerging on the mammalian in vivo absorption of engineered NMs composed of chemicals with a range of properties, including metal, mineral, biochemical macromolecules, and lipid-based entities. In vitro and in silico fluid incubation data has also provided some evidence of changes in particle stability, aggregation, and surface properties following interaction with luminal factors present in the GI tract. The variables include physical forces, osmotic concentration, pH, digestive enzymes, other food, and endogenous biochemicals, and commensal microbes. Further research is required to fill remaining data gaps on the effects of these parameters on NM integrity, physicochemical properties, and GI absorption. Knowledge of the most influential luminal parameters will be essential when developing models of the GI tract to quantify the percent absorption of food-relevant engineered NMs for risk assessment.

A review and perspective of existing research on the release of nanomaterials from solid nanocomposites.

Advances in adding nanomaterials to various matrices have occurred in tandem with the identification of potential hazards associated with exposure to pure forms of nanomaterials. We searched multiple research publication databases and found that, relative to data generated on potential nanomaterial hazards or exposures, very little attention has focused on understanding the potential and conditions for release of nanomaterials from nanocomposites. However, as a prerequisite to exposure studying release is necessary to inform risk assessments. We identified fifty-four studies that specifically investigated the release of nanomaterials, and review them in the following release scenario groupings: machining, weathering, washing, contact and incineration. While all of the identified studies provided useful information, only half were controlled experiments. Based on these data, the debris released from solid, non-food nanocomposites contains in varying frequencies, a mixture of four types of debris. Most frequently identified are (1) particles of matrix alone, and slightly less often, the (2) matrix particles exhibit the nanomaterial partially or fully embedded; far less frequently is (3) the added nanomaterial entirely dissociated from the matrix identified: and most rare are (4) dissolved ionic forms of the added nanomaterial. The occurrence of specific debris types appeared to be dependent on the specific release scenario and environment. These data highlight that release from nanocomposites can take multiple forms and that additional research and guidance would be beneficial, allowing for more consistent characterization of the release potential of nanomaterials. In addition, these data support calls for method validation and standardization, as well as understanding how laboratory release scenarios relate to real-world conditions. Importantly, as risk is considered to be a function of the inherent hazards of a substance and the actual potential for exposure, data on nanomaterial release dynamics and debris composition from commercially relevant nanocomposites are a valuable starting point for consideration in fate and transport modeling, exposure assessment, and risk assessment frameworks for nanomaterials.

Measurement of nanomaterials in foods: integrative consideration of challenges and future prospects.

The risks and benefits of nanomaterials in foods and food contact materials receive conflicting international attention across expert stakeholder groups as well as in news media coverage and published research. Current nanomaterial characterization is complicated by the lack of accepted approaches to measure exposure-relevant occurrences of suspected nanomaterials in food and by broad definitions related to food processing and additive materials. Therefore, to improve understanding of risk and benefit, analytical methods are needed to identify what materials, new or traditional, are "nanorelevant" with respect to biological interaction and/or uptake during alimentary tract transit. Challenges to method development in this arena include heterogeneity in nanomaterial composition and morphology, food matrix complexity, alimentary tract diversity, and analytical method limitations. Clear problem formulation is required to overcome these and other challenges and to improve understanding of biological fate in facilitating the assessment of nanomaterial safety or benefit, including sampling strategies relevant to food production/consumption and alimentary tract transit. In this Perspective, we discuss critical knowledge gaps that must be addressed so that measurement methods can better inform risk management and public policy.

Engineered Nanoscale Food Ingredients: Evaluation of Current Knowledge on Material Characteristics Relevant to Uptake from the Gastrointestinal Tract.

The NanoRelease Food Additive project developed a catalog to identify potential engineered nanomaterials (ENMs) used as ingredients, using various food-related databases. To avoid ongoing debate on defining the term nanomaterial, NanoRelease did not use any specific definition other than the ingredient is not naturally part of the food chain, and its dimensions are measured in the nanoscale. Potential nanomaterials were categorized based on physical similarity; analysis indicated that the range of ENMs declared as being in the food chain was limited. Much of the catalog's information was obtained from product labeling, likely resulting in both underreporting (inconsistent or absent requirements for labeling) and/or overreporting (inability to validate entries, or the term nano was used, although no ENM material was present). Three categories of ingredients were identified: emulsions, dispersions, and their water-soluble powdered preparations (including lipid-based structures); solid encapsulates (solid structures containing an active material); and metallic or other inorganic particles. Although much is known regarding the physical/chemical properties for these ingredient categories, it is critical to understand whether these properties undergo changes following their interaction with food matrices during preparation and storage. It is also important to determine whether free ENMs are likely to be present within the gastrointestinal tract and whether uptake of ENMs may occur in their nanoform physical state. A practical decision-making scheme was developed to help manage testing requirements.

Insights and perspectives on emerging inputs to weight of evidence determinations for food safety: workshop proceedings.

This workshop aimed to elucidate the contribution of computational and emerging in vitro methods to the weight of evidence used by risk assessors in food safety assessments. The following issues were discussed: using in silico and high-throughput screening (HTS) data to confirm the safety of approved food ingredients, applying in silico and HTS data in the process of assessing the safety of a new food ingredient, and utilizing in silico and HTS data in communicating the safety of food ingredients while enhancing the public's trust in the food supply. Perspectives on integrating computational modeling and HTS assays as well as recommendations for optimizing predictive methods for risk assessment were also provided. Given the need to act quickly or proceed cautiously as new data emerge, this workshop also focused on effectively identifying a path forward in communicating in silico and in vitro data.

Determining the applicability of threshold of toxicological concern approaches to substances found in foods.

Threshold of Toxicological Concern (TTC) decision-support methods present a pragmatic approach to using data from well-characterized chemicals and protective estimates of exposure in a stepwise fashion to inform decisions regarding low-level exposures to chemicals for which few data exist. It is based on structural and functional categorizations of chemicals derived from decades of animal testing with a wide variety of chemicals. Expertise is required to use the TTC methods, and there are situations in which its use is clearly inappropriate or not currently supported. To facilitate proper use of the TTC, this paper describes issues to be considered by risk managers when faced with the situation of an unexpected substance in food. Case studies are provided to illustrate the implementation of these considerations, demonstrating the steps taken in deciding whether it would be appropriate to apply the TTC approach in each case. By appropriately applying the methods, employing the appropriate scientific expertise, and combining use with the conservative assumptions embedded within the derivation of the thresholds, the TTC can realize its potential to protect public health and to contribute to efficient use of resources in food safety risk management.

The uncertainty of nanotoxicology: report of a Society for Risk Analysis Workshop.

A September 2008 workshop sponsored by the Society for Risk Analysis(() (1) ()) on risk assessment methods for nanoscale materials explored "nanotoxicology" in risk assessment. A general conclusion of the workshop was that, while research indicates that some nanoscale materials are toxic, the information presented at the workshop does not indicate the need for a conceptually different approach for risk assessment on nanoscale materials, compared to other materials. However, the toxicology discussions did identify areas of uncertainty that present a challenge for the assessment of nanoscale materials. These areas include novel metrics, characterizing multivariate dynamic mixtures, identification of toxicologically relevant properties and "impurities" for nanoscale characteristics, and characterizing persistence, toxicokinetics, and weight of evidence in consideration of the dynamic nature of the mixtures. The discussion also considered "nanomaterial uncertainty factors" for health risk values like the Environmental Protection Agency's reference dose (RfD). Similar to the general opinions for risk assessment, participants expressed that completing a data set regarding toxicity, or extrapolation between species, sensitive individuals, or durations of exposure, were not qualitatively different considerations for nanoscale materials in comparison to all chemicals, and therefore, a "nanomaterial uncertainty factor" for all nanomaterials does not seem appropriate. However, the quantitative challenges may require new methods and approaches to integrate the information and the uncertainty.

Nano risk analysis: advancing the science for nanomaterials risk management.

Scientists, activists, industry, and governments have raised concerns about health and environmental risks of nanoscale materials. The Society for Risk Analysis convened experts in September 2008 in Washington, DC to deliberate on issues relating to the unique attributes of nanoscale materials that raise novel concerns about health risks. This article reports on the overall themes and findings of the workshop, uncovering the underlying issues for each of these topics that become recurring themes. The attributes of nanoscale particles and other nanomaterials that present novel issues for risk analysis are evaluated in a risk analysis framework, identifying challenges and opportunities for risk analysts and others seeking to assess and manage the risks from emerging nanoscale materials and nanotechnologies. Workshop deliberations and recommendations for advancing the risk analysis and management of nanotechnologies are presented.

Meta-analysis of dioxin cancer dose response for three occupational cohorts.

This article presents a meta analysis of data from three cohorts occupationally exposed to TCDD and related compounds. A statistically significant (p = 0.02) trend was found in total cancer mortality with increasing dioxin exposure. The trend tests show an increase in total cancer at cumulative TEQ (unit of measurement for TCDD-like compounds that is defined as the amount of TCDD that would produce the same toxicity as a mixture of TCDD-like compounds) serum levels that would result from lifetime intake of 7 pg TEQ/kg body weight/day, with no increase at 6 pg/kg/day. A linear dose response provided a good fit to the combined data and predicted an ED(01) (dioxin exposure resulting in a 0.01 increase in lifetime risk of cancer mortality) of 45 pg/kg/day (95% confidence interval, 21-324). The U.S. Environmental Protection Agency estimates that current lifetime human exposures to dioxin average approximately 1 pg/kg/day (99% percentile: 3 pg/kg/day). Although it appears unlikely that current exposures through foods would reach either 7 pg/kg/day or the ED(01), our analysis argues for careful consideration of the upper ranges of long-term average exposures for dioxins.

Blood mercury levels in US children and women of childbearing age, 1999-2000.

CONTEXT: Humans are exposed to methylmercury, a well-established neurotoxin, through fish consumption. The fetus is most sensitive to the adverse effects of exposure. The extent of exposure to methylmercury in US women of reproductive age is not known. OBJECTIVE: To describe the distribution of blood mercury levels in US children and women of childbearing age and the association with sociodemographic characteristics and fish consumption. DESIGN AND SETTING: The 1999-2000 data from the National Health and Nutrition Examination Survey, a cross-sectional survey of the noninstitutionalized US population. PARTICIPANTS: In 1999-2000, 1250 children aged 1 to 5 years and 2314 women aged 16 to 49 years were selected to participate in the survey. Household interviews, physical examinations, and blood mercury levels assessments were performed on 705 children (56% response rate) and 1709 women (74% response rate). MAIN OUTCOME MEASURE: Blood concentration of total mercury. RESULTS: Blood mercury levels were approximately 3-fold higher in women compared with children. The geometric mean concentration of total blood mercury was 0.34 micro g/L (95% confidence interval [CI], 0.30-0.39 microg/L) in children and 1.02 microg/L (95% CI, 0.85-1.20 microg/L) in women. Geometric mean mercury levels were almost 4-fold higher among women who ate 3 or more servings of fish in the past 30 days compared with women who ate no fish in that period (1.94 microg/L vs 0.51 microg/L; P<.001). CONCLUSIONS: Measures of mercury exposure in women of childbearing age and young children generally fall below levels of concern. However, approximately 8% of women had concentrations higher than the US Environmental Protection Agency's recommended reference dose (5.8 microg/L), below which exposures are considered to be without adverse effects. Women who are pregnant or who intend to become pregnant should follow federal and state advisories on consumption of fish.