Safe(r) by design guidelines for the nanotechnology industry.

Expectations for safer and sustainable chemicals and products are growing to comply with the United Nations and European strategies for sustainability. The application of Safe(r) by Design (SbD) in nanotechnology implies an iterative process where functionality, human health and safety, environmental and economic impact and cost are assessed and balanced as early as possible in the innovation process and updated at each step. The EU H2020 NanoReg2 project was the first European project to implement SbD in six companies handling and/or manufacturing nanomaterials (NMs) and nano-enabled products (NEP). The results from this experience have been used to develop these guidelines on the practical application of SbD. The SbD approach foresees the identification, estimation, and reduction of human and environmental risks as early as possible in the development of a NM or NEP, and it is based on three pillars: (i) safer NMs and NEP; (ii) safer use and end of life and (iii) safer industrial production. The presented guidelines include a set of information and tools that will help deciding at each step of the innovation process whether to continue, apply SbD measures or carry out further tests to reduce uncertainty. It does not intend to be a prescriptive protocol where all suggested steps have to be followed to achieve a SbD NM/NEP or process. Rather, the guidelines are designed to identify risks at an early state and information to be considered to identify those risks. Each company adapts the approach to its specific needs and circumstances as company decisions influence the way forward.

Integrative approach in a safe by design context combining risk, life cycle and socio-economic assessment for safer and sustainable nanomaterials.

Moving towards safe and sustainable innovations is an international policy ambition. In the on-hand manuscript, a concept combining safe by design and sustainability was implemented through the integration of human and environmental risk assessment, life cycle assessment as well as an assessment of the economic viability. The result is a nested and iterative process in form of a decision tree that integrates these three elements in order to achieve sustainable, safe and competitive materials, products or services. This approach, embedded into the stage-gate-model for safe by design, allows to reduce the uncertainty related to the assessment of risks and impacts by improving the quality of the data collected along each stage. In the second part of the manuscript, the application is shown for a case study dealing with the application of nanoparticles for Li-Ion batteries. One of the general conclusions out of this case study is that data gaps are a key aspect in view of the reliability of the results.

An integrated pathway based on in vitro data for the human hazard assessment of nanomaterials.

In line with the 3R concept, nanotoxicology is shifting from a phenomenological to a mechanistic approach based on in vitro and in silico methods, with a consequent reduction in animal testing. Risk Assessment (RA) and Life Cycle Assessment (LCA) methodologies, which traditionally rely on in vivo toxicity studies, will not be able to keep up with the pace of development of new nanomaterials unless they adapt to use this new type of data. While tools and models are already available and show a great potential for future use in RA and LCA, currently none is able alone to quantitatively assess human hazards (i.e. calculate chronic NOAEL or ED50 values). By highlighting which models and approaches can be used in a quantitative way with the available knowledge and data, we propose an integrated pathway for the use of in vitro data in RA and LCA. Starting with the characterization of nanoparticles' properties, the pathway then investigates how to select relevant in vitro human data, and how to bridge in vitro dose-response relationships to in vivo effects. If verified, this approach would allow RA and LCA to stir up the development of nanotoxicology by giving indications about the data and quality requirements needed in risk methodologies.

Relative potency factor approach enables the use of in vitro information for estimation of human effect factors for nanoparticle toxicity in life-cycle impact assessment.

The major theme of the NRC report "Toxicity Testing in the Twenty-first Century" is to replace animal testing by using alternative in vitro methods. Therefore, it can be expected that in the future in vivo data will be replaced with in vitro data. Hence, there is a need for new strategies to make use of the increasing amount of in vitro data when developing human toxicological effect factors (HEF) to characterize the impact category of human toxicity in life cycle assessment (LCA). Here, we present a new approach for deriving HEF for manufactured nanomaterials (MNMs) based on the combined use of in vitro toxicity data and a relative potency factor (RPF) approach. In vitro toxicity tests with nano-CuO, nano-Ag and nano-ZnO and their corresponding ions were performed on THP-1, CaCo-2 and Hep-G2 cell lines. The ratio of the here calculated EC50 of the ionic form and the nanoform corresponds to the Relative Potency Factor (RPF). Using this approach, HEFs (case/kgintake) for the aforementioned nanoparticles were obtained. Non-carcinogenic HEFs (case/kgintake) for exposure via ingestion of 5.9E-01, 7.5E-03 and 2.5 E-02 were calculated for nano-Ag, nano-CuO and nano-ZnO, respectively. The HEF values here proposed were compared with HEF values extrapolated from in vivo toxicity data reported in the literature. The here presented procedure is the most appropriate approximation currently available for using in vitro toxicity data on MNM for application in the field of LCIA.

Freshwater ecotoxicity characterisation factor for metal oxide nanoparticles: a case study on titanium dioxide nanoparticle.

The Life Cycle Assessment (LCA) methodology is widely applied in several industrial sectors to evaluate the environmental performance of processes, products and services. Recently, several reports and studies have emphasized the importance of LCA in the field of engineered nanomaterials. However, to date only a few LCA studies on nanotechnology have been carried out, and fewer still have assessed aspects relating to ecotoxicity. This is mainly due to the lack of knowledge in relation on human and environmental exposure and effect of engineered nanoparticles (ENPs). This bottleneck is continued when performing Life Cycle Impact Assessment, where characterization models and consequently characterization factors (CFs) for ENPs are missing. This paper aims to provide the freshwater ecotoxicity CF for titanium dioxide nanoparticles (nano-TiO2). The USEtox model has been selected as a characterisation model. An adjusted multimedia fate model has been developed which accounts for nano-specific fate process descriptors (i.e. sedimentation, aggregation with suspended particle matter, etc.) to estimate the fate of nano-TiO2 in freshwater. A literature survey of toxicity tests performed on freshwater organism representative of multiple trophic levels was conducted, including algae, crustaceans and fish in order to collect relevant EC50 values. Then, the toxic effect of nano-TiO2 was computed on the basis of the HC50 value. Thus, following the principle of USEtox model and accounting for nano-specific descriptors a CF for the toxic impact of freshwater ecotoxicity of 0.28 PAFdaym(3)kg(-1) is proposed.

Does the exposure mode to ENPs influence their toxicity to aquatic species? A case study with TiO2 nanoparticles and Daphnia magna.

Recent studies suggest that the ecotoxicity of engineered nanoparticles (ENPs) is dependent upon the treatment of ENPs in suspensions (e.g. sonication or use of solvents) and on the mode of exposure to test organisms. We conducted several bioassays with Daphnia magna in order to determine how adverse effects of TiO2 nanoparticles (n-TiO2) are influenced by experimental set-up. Several treatments were applied, including three test media, several treatments of n-TiO2 suspensions (stirring, sonication) and different exposure modes (exposure duration and volume of test suspension). No adverse effects were observed when D. magna were exposed to 50 mL of suspension, regardless of TiO2 concentration (up to 250 mg/L) and exposure duration. Conversely, adverse effects were observed when D. magna were exposed to 2 mL of suspension for 96 h with a 50 % effect concentration EC50 values ranging from 32 mg/L to 82 mg/L. Test media had no significant influence on the outcome of all treatments. For a better mechanistic understanding of the experimental set-up at which adverse effects were observed, the particle size of n-TiO2 in the test media was characterized throughout the test duration. These measurements revealed a fast and strong agglomeration with a secondary particle size in the order of magnitude of micrometers. Our study describes how the effects of n-TiO2 on D .magna are influenced by the duration of exposure and volume of media, highlighting the need for standardization of experimental methods.