Roadmap towards safe and sustainable advanced and innovative materials. (Outlook for 2024-2030).

The adoption of innovative advanced materials holds vast potential, contingent upon addressing safety and sustainability concerns. The European Commission advocates the integration of Safe and Sustainable by Design (SSbD) principles early in the innovation process to streamline market introduction and mitigate costs. Within this framework, encompassing ecological, social, and economic factors is paramount. The NanoSafety Cluster (NSC) delineates key safety and sustainability areas, pinpointing unresolved issues and research gaps to steer the development of safe(r) materials. Leveraging FAIR data management and integration, alongside the alignment of regulatory aspects, fosters informed decision-making and innovation. Integrating circularity and sustainability mandates clear guidance, ensuring responsible innovation at every stage. Collaboration among stakeholders, anticipation of regulatory demands, and a commitment to sustainability are pivotal for translating SSbD into tangible advancements. Harmonizing standards and test guidelines, along with regulatory preparedness through an exchange platform, is imperative for governance and market readiness. By adhering to these principles, the effective and sustainable deployment of innovative materials can be realized, propelling positive transformation and societal acceptance.

Indoor paint life cycle particle release: Safer-by-design products and the importance of choosing the right formula.

In 2020, the European Commission published a regulation that states all producers of white paints containing titanium dioxide (TiO2) must provide a warning label on their products. Exposure during the production and application of products containing TiO2 can be harmful, and therefore these products must be labeled as "may cause cancer." The paint industry is a major user of TiO2 pigment. This study focuses on pigment release from three TiO2-based paints and discusses the effect of paint formulation, more precisely the Pigment Volume Concentration (PVC), to predict TiO2 pigment release from the paints during a simulated use phase and at the end of life (EoL). The use phase considered mild abrasion of painted panels that simulated cleaning or touching. The EoL phase was studied using leaching tests simulating landfill disposal. TiO2 release during both activities was evident with a high discrepancy between the three paints. While dry rubbing was similar for all paints, activities involving water present a high release link to paint matrix degradation. The paint pigment volume concentration and the paint permeability determines the TiO2 release during wet rubbing and leaching. This work represents an attempt to identify the paint permeability as a matrix-related parameter to predict TiO2 release and a way to use of this parameter to develop safer products.

Strategies for Enhancing Plant Immunity and Resilience Using Nanomaterials for Sustainable Agriculture.

Research on plant-nanomaterial interactions has greatly advanced over the past decade. One particularly fascinating discovery encompasses the immunomodulatory effects in plants. Due to the low doses needed and the comparatively low toxicity of many nanomaterials, nanoenabled immunomodulation is environmentally and economically promising for agriculture. It may reduce environmental costs associated with excessive use of chemical pesticides and fertilizers, which can lead to soil and water pollution. Furthermore, nanoenabled strategies can enhance plant resilience against various biotic and abiotic stresses, contributing to the sustainability of agricultural ecosystems and the reduction of crop losses due to environmental factors. While nanoparticle immunomodulatory effects are relatively well-known in animals, they are still to be understood in plants. Here, we provide our perspective on the general components of the plant's immune system, including the signaling pathways, networks, and molecules of relevance for plant nanomodulation. We discuss the recent scientific progress in nanoenabled immunomodulation and nanopriming and lay out key avenues to use plant immunomodulation for agriculture. Reactive oxygen species (ROS), the mitogen-activated protein kinase (MAPK) cascade, and the calcium-dependent protein kinase (CDPK or CPK) pathway are of particular interest due to their interconnected function and significance in the response to biotic and abiotic stress. Additionally, we underscore that understanding the plant hormone salicylic acid is vital for nanoenabled applications to induce systemic acquired resistance. It is suggested that a multidisciplinary approach, incorporating environmental impact assessments and focusing on scalability, can expedite the realization of enhanced crop yields through nanotechnology while fostering a healthier environment.

Nanosafety: a Perspective on Nano-Bio Interactions.

Engineered nanomaterials offer numerous benefits to society ranging from environmental remediation to biomedical applications such as drug or vaccine delivery as well as clean and cost-effective energy production and storage, and the promise of a more sustainable way of life. However, as nanomaterials of increasing sophistication enter the market, close attention to potential adverse effects on human health and the environment is needed. Here a critical perspective on nanotoxicological research is provided; the authors argue that it is time to leverage the knowledge regarding the biological interactions of nanomaterials to achieve a more comprehensive understanding of the human health and environmental impacts of these materials. Moreover, it is posited that nanomaterials behave like biological entities and that they should be regulated as such.

Environmental and Health Impacts of Graphene and Other Two-Dimensional Materials: A Graphene Flagship Perspective.

Two-dimensional (2D) materials have attracted tremendous interest ever since the isolation of atomically thin sheets of graphene in 2004 due to the specific and versatile properties of these materials. However, the increasing production and use of 2D materials necessitate a thorough evaluation of the potential impact on human health and the environment. Furthermore, harmonized test protocols are needed with which to assess the safety of 2D materials. The Graphene Flagship project (2013-2023), funded by the European Commission, addressed the identification of the possible hazard of graphene-based materials as well as emerging 2D materials including transition metal dichalcogenides, hexagonal boron nitride, and others. Additionally, so-called green chemistry approaches were explored to achieve the goal of a safe and sustainable production and use of this fascinating family of nanomaterials. The present review provides a compact survey of the findings and the lessons learned in the Graphene Flagship.

A comparison of dermal toxicity models; assessing suitability for safe(r)-by-design decision-making and for screening nanomaterial hazards.

The objective of Safe-by-Design (SbD) is to support the development of safer products and production processes, and enable safe use throughout a materials' life cycle; an intervention at an early stage of innovation can greatly benefit industry by reducing costs associated with the development of products later found to elicit harmful effects. Early hazard screening can support this process, and is needed for all of the expected nanomaterial exposure routes, including inhalation, ingestion and dermal. In this study, we compare in vitro and ex vivo cell models that represent dermal exposures (including HaCaT cells, primary keratinocytes, and reconstructed human epidermis (RhE)), and when possible consider these in the context of regulatory accepted OECD TG for in vitro dermal irritation. Various benchmark nanomaterials were used to assess markers of cell stress in each cell model. In addition, we evaluated different dosing strategies that have been used when applying the OECD TG for dermal irritation in assessment of nanomaterials, and how inconsistencies in the approach used can have considerable impact of the conclusions made. Although we could not demonstrate alignment of all models used, there was an indication that the simpler in vitro cell model aligned more closely with RhE tissue than ex vivo primary keratinocytes, supporting the use of HaCaT cells for screening of dermal toxicity of nanomaterials and in early-stage SbD decision-making.

Advanced optical imaging for the rational design of nanomedicines.

Despite the enormous potential of nanomedicines to shape the future of medicine, their clinical translation remains suboptimal. Translational challenges are present in every step of the development pipeline, from a lack of understanding of patient heterogeneity to insufficient insights on nanoparticle properties and their impact on material-cell interactions. Here, we discuss how the adoption of advanced optical microscopy techniques, such as super-resolution optical microscopies, correlative techniques, and high-content modalities, could aid the rational design of nanocarriers, by characterizing the cell, the nanomaterial, and their interaction with unprecedented spatial and/or temporal detail. In this nanomedicine arena, we will discuss how the implementation of these techniques, with their versatility and specificity, can yield high volumes of multi-parametric data; and how machine learning can aid the rapid advances in microscopy: from image acquisition to data interpretation.

Learning from Safe-by-Design for Safe-and-Sustainable-by-Design: Mapping the current landscape of Safe-by-Design reviews, case studies, and frameworks.

With the introduction of the European Commission's "Safe and Sustainable-by-Design" (SSbD) framework, the interest in understanding the implications of safety and sustainability assessments of chemicals, materials, and processes at early-innovation stages has skyrocketed. Our study focuses on the "Safe-by-Design" (SbD) approach from the nanomaterials sector, which predates the SSbD framework. In this assessment, SbD studies have been compiled and categorized into reviews, case studies, and frameworks. Reviews of SbD tools have been further classified as quantitative, qualitative, or toolboxes and repositories. We assessed the SbD case studies and classified them into three categories: safe(r)-by-modeling, safe(r)-by-selection, or safe(r)-by-redesign. This classification enabled us to understand past SbD work and subsequently use it to define future SSbD work so as to avoid confusion and possibilities of "SSbD-washing" (similar to greenwashing). Finally, the preexisting SbD frameworks have been studied and contextualized against the SSbD framework. Several key recommendations for SSbD based on our analysis can be made. Knowledge gained from existing approaches such as SbD, green and sustainable chemistry, and benign-by-design approaches needs to be preserved and effectively transferred to SSbD. Better incorporation of chemical and material functionality into the SSbD framework is required. The concept of lifecycle thinking and the stage-gate innovation model need to be reconciled for SSbD. The development of high-throughput screening models is critical for the operationalization of SSbD. We conclude that the rapid pace of both SbD and SSbD development necessitates a regular mapping of the newly published literature that is relevant to this field.

Modulating the toxicity of engineered nanoparticles by controlling protein corona formation: Recent advances and future prospects.

With the rapid development and widespread application of engineered nanoparticles (ENPs), understanding the fundamental interactions between ENPs and biological systems is essential to assess and predict the fate of ENPs in vivo. When ENPs are exposed to complex physiological environments, biomolecules quickly and inevitably adsorb to ENPs to form a biomolecule corona, such as a protein corona (PC). The formed PC has a significant effect on the physicochemical properties of ENPs and gives them a brand new identity in the biological environment, which determines the subsequent ENP-cell/tissue/organ interactions. Controlling the formation of PCs is therefore of utmost importance to accurately predict and optimize the behavior of ENPs within living organisms, as well as ensure the safety of their applications. In this review, we provide an overview of the fundamental aspects of the PC, including the formation mechanism, composition, and frequently used characterization techniques. We comprehensively discuss the potential impact of the PC on ENP toxicity, including cytotoxicity, immune response, and so on. Additionally, we summarize recent advancements in manipulating PC formation on ENPs to achieve the desired biological outcomes. We further discuss the challenges and prospects, aiming to provide valuable insights for a better understanding and prediction of ENP behaviors in vivo, as well as the development of low-toxicity ENPs.

How safe are nanoscale metal-organic frameworks?

Owing to the size scales that can be accessed, the nanoscale has opened doors to new physical and chemical properties, not seen in the bulk. These properties are leveraged by nanomaterials (NMs) across a plethora of applications. More recently, nanoscale metal-organic frameworks (nMOFs) have witnessed explosive growth due to the modularity of their chemical constituents, the ability to modify their composition and structure, and exceptional properties such as permanent porosity and high surface areas. These properties have prompted the investigation of these materials for applications in biological and environmental contexts. However, one aspect that is often ignored in these discussions is their safety at a nanoscale. In this mini review, we aim to initiate a discussion on the safety and toxicity of nMOFs, drawing parallels with the existing guidelines and literature on the safety of inorganic NMs. We first describe why nMOFs are of considerable interest to the scientific community followed by a discussion on routes through which they can be exposed to the environment and living organisms, particularly shedding light on their transformation mechanisms. The review also discusses the factors affecting toxicity of nMOFs, such as their size, shape, morphology, and composition. We briefly highlight potential mechanisms of toxicity and conclude with describing the need to transition towards data-intensive computational approaches such as machine learning to establish nMOFs as credible materials for their envisioned applications.

Release and toxicity assessment of carbon nanomaterial reinforced polymers during the use and end-of-life phases: A comparative review.

The research on carbon-based nanomaterial (C-NM) composites has increased in the last two decades. This family of functional materials shows outstanding mechanical, thermal and electrical properties, and are being used in a variety of applications. An important challenge remains before C-NM can be fully integrated in our production industries and our lives: to assess the release of debris during production, use, and misuse of composites and the effect they may have on the environment and on human health. During their lifecycle, composites materials can be subjected to a variety of stresses which may release particles from the macroscopic range to the nanoscale. In this review, the release of debris due to abrasion, weathering and combustion as well as their toxicity is evaluated for the three most used C-NM: Carbon Black, Carbon Nanotubes and Graphene-related materials. The goal is to stimulate a Safe-By-Design approach by guiding the selection of carbon nano-fillers for specific applications based of safety and performance.

Assessing the Fate of Superparamagnetic Iron Oxide Nanoparticles Carrying Usnic Acid as Chemical Cargo on the Soil Microbial Community.

In the present study we evaluate the effect of superparamagnetic iron oxide nanoparticles (SPIONs) carrying usnic acid (UA) as chemical cargo on the soil microbial community in a dystrophic red latosol (oxysol). Herein, 500 ppm UA or SPIONs-framework carrying UA were diluted in sterile ultrapure deionized water and applied by hand sprayer on the top of the soil. The experiment was conducted in a growth chamber at 25 °C, with a relative humidity of 80% and a 16 h/8 h light-dark cycle (600 lx light intensity) for 30 days. Sterile ultrapure deionized water was used as the negative control; uncapped and oleic acid (OA) capped SPIONs were also tested to assess their potential effects. Magnetic nanostructures were synthesized by a coprecipitation method and characterized by scanning and transmission electron microscopy (SEM and TEM), X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), zeta potential, hydrodynamic diameter, magnetic measurements, and release kinetics of chemical cargo. Uncapped and OA-capped SPIONs did not significantly affect soil microbial community. Our results showed an impairment in the soil microbial community exposed to free UA, leading to a general decrease in negative effects on soil-based parameters when bioactive was loaded into the nanoscale magnetic carrier. Besides, compared to control, the free UA caused a significant decrease in microbial biomass C (39%), on the activity of acid protease (59%), and acid phosphatase (23%) enzymes, respectively. Free UA also reduced eukaryotic 18S rRNA gene abundance, suggesting a major impact on fungi. Our findings indicate that SPIONs as bioherbicide nanocarriers can reduce the negative impacts on soil. Therefore, nanoenabled biocides may improve agricultural productivity, which is important for food security due to the need of increasing food production.

Exploring the Impact of Tensions in Stakeholder Norms on Designing for Value Change: The Case of Biosafety in Industrial Biotechnology.

Synthetic biologists design and engineer organisms for a better and more sustainable future. While the manifold prospects are encouraging, concerns about the uncertain risks of genome editing affect public opinion as well as local regulations. As a consequence, biosafety and associated concepts, such as the Safe-by-design framework and genetic safeguard technologies, have gained notoriety and occupy a central position in the conversation about genetically modified organisms. Yet, as regulatory interest and academic research in genetic safeguard technologies advance, the implementation in industrial biotechnology, a sector that is already employing engineered microorganisms, lags behind. The main goal of this work is to explore the utilization of genetic safeguard technologies for designing biosafety in industrial biotechnology. Based on our results, we posit that biosafety is a case of a changing value, by means of further specification of how to realize biosafety. Our investigation is inspired by the Value Sensitive Design framework, to investigate scientific and technological choices in their appropriate social context. Our findings discuss stakeholder norms for biosafety, reasonings about genetic safeguards, and how these impact the practice of designing for biosafety. We show that tensions between stakeholders occur at the level of norms, and that prior stakeholder alignment is crucial for value specification to happen in practice. Finally, we elaborate in different reasonings about genetic safeguards for biosafety and conclude that, in absence of a common multi-stakeholder effort, the differences in informal biosafety norms and the disparity in biosafety thinking could end up leading to design requirements for compliance instead of for safety.

Preliminary Toxicological Analysis in a Safe-by-Design and Adverse Outcome Pathway-Driven Approach on Different Silver Nanoparticles: Assessment of Acute Responses in A549 Cells.

Silver nanoparticles (Ag NPs) are among the most widely used metal-based nanomaterials (NMs) and their applications in different products, also as antibacterial additives, are increasing. In the present manuscript, according to an adverse outcome pathway (AOP) approach, we tested two safe-by-design (SbD) newly developed Ag NPs coated with hydroxyethyl cellulose (HEC), namely AgHEC powder and AgHEC solution. These novel Ag NPs were compared to two reference Ag NPs (naked and coated with polyvinylpyrrolidone-PVP). Cell viability, inflammatory response, reactive oxygen species, oxidative DNA damage, cell cycle, and cell-particle interactions were analyzed in the alveolar in vitro model, A549 cells. The results show a different toxicity pattern of the novel Ag NPs compared to reference NPs and that between the two novel NPs, the AgHEC solution is the one with the lower toxicity and to be further developed within the SbD framework.

The State of the Art and Challenges of In Vitro Methods for Human Hazard Assessment of Nanomaterials in the Context of Safe-by-Design.

The Safe-by-Design (SbD) concept aims to facilitate the development of safer materials/products, safer production, and safer use and end-of-life by performing timely SbD interventions to reduce hazard, exposure, or both. Early hazard screening is a crucial first step in this process. In this review, for the first time, commonly used in vitro assays are evaluated for their suitability for SbD hazard testing of nanomaterials (NMs). The goal of SbD hazard testing is identifying hazard warnings in the early stages of innovation. For this purpose, assays should be simple, cost-effective, predictive, robust, and compatible. For several toxicological endpoints, there are indications that commonly used in vitro assays are able to predict hazard warnings. In addition to the evaluation of assays, this review provides insights into the effects of the choice of cell type, exposure and dispersion protocol, and the (in)accurate determination of dose delivered to cells on predictivity. Furthermore, compatibility of assays with challenging advanced materials and NMs released from nano-enabled products (NEPs) during the lifecycle is assessed, as these aspects are crucial for SbD hazard testing. To conclude, hazard screening of NMs is complex and joint efforts between innovators, scientists, and regulators are needed to further improve SbD hazard testing.

Secured Nanosynthesis-Deposition Aerosol Process for Composite Thin Films Incorporating Highly Dispersed Nanoparticles.

Application of nanocomposites in daily life requires not only small nanoparticles (NPs) well dispersed in a matrix, but also a manufacturing process that is mindful of the operator and the environment. Avoiding any exposure to NPs is one such way, and direct liquid reaction-injection (DLRI) aims to fulfill this need. DLRI is based on the controlled in situ synthesis of NPs from the decomposition of suitable organometallic precursors in conditions that are compatible with a pulsed injection mode of an aerosol into a downstream process. Coupled with low-pressure plasma, DLRI produces nanocomposite with homogeneously well-dispersed small nanoparticles that in the particular case of ZnO-DLC nanocomposite exhibit unique properties. DLRI favorably compares with the direct liquid injection of ex situ formed NPs. The exothermic hydrolysis reaction of the organometallic precursor at the droplet-gas interface leads to the injection of small and highly dispersed NPs and, consequently, the deposition of fine and controlled distribution in the nanocomposite. The scope of DLRI nanosynthesis has been extended to several metal oxides such as zinc, tin, tungsten, and copper to generalize the concept. Hence, DLRI is an attractive method to synthesize, inject, and deposit nanoparticles and meets the prevention and atom economy requirements of green chemistry.

Approaches to implement safe by design in early product design through combining risk assessment and Life Cycle Assessment.

The Safe by Design (SbD) concept aims to ensure the production, use and disposal of materials and products safely. While there is a growing interest in the potential of SbD to support policy commitments, such as the EU Green Deal and the Circular Economy Action Plan in Europe, methodological approaches and practical guidelines on SbD are, however, largely missing. The combined use of Life Cycle Assessment (LCA) and Risk Assessment (RA) is considered suitable to operationalize SbD over the whole life-cycle of a product. Here, we explore the potential of the combined use of LCA and RA at Technological Readiness Level (TRL) 1-6. We perform a review of the literature presenting and/or developing approaches that combine LCA and RA at early stages of product design. We identify that basic early-on-evaluations of safety (e.g., apply lifecycle thinking to assess risk hotspots, avoid use of hazardous chemicals, minimize other environmental impacts from chemicals) are more common, while more complex assessments (e.g., ex-ante LCA, control banding, predictive (eco)toxicology) require specialized expertise. The application of these simplified approaches and guidelines aims to avoid some obvious sources of risks and impacts at early stages. Critical gaps need to be addressed for wider application of SbD, including more studies in the product design context, developing tools and databases containing collated information on risk, greater collaboration between RA/LCA researchers and companies, and policy discussion on the expansion from SbD to Safe and Sustainable by Design (SSbD).

Quantitative adverse outcome pathway (qAOP) using bayesian network model on comparative toxicity of multi-walled carbon nanotubes (MWCNTs): safe-by-design approach.

While the various physicochemical properties of engineered nanomaterials influence their toxicities, their understanding is still incomplete. A predictive framework is required to develop safe nanomaterials, and a Bayesian network (BN) model based on adverse outcome pathway (AOP) can be utilized for this purpose. In this study, to explore the applicability of the AOP-based BN model in the development of safe nanomaterials, a comparative study was conducted on the change in the probability of toxicity pathways in response to changes in the dimensions and surface functionalization of multi-walled carbon nanotubes (MWCNTs). Based on the results of our previous study, we developed an AOP leading to cell death, and the experimental results were collected in human liver cells (HepG2) and bronchial epithelium cells (Beas-2B). The BN model was trained on these data to identify probabilistic causal relationships between key events. The results indicated that dimensions were the main influencing factor for lung cells, whereas -OH or -COOH surface functionalization and aspect ratio were the main influencing factors for liver cells. Endoplasmic reticulum stress was found to be a more sensitive pathway for dimensional changes, and oxidative stress was a more sensitive pathway for surface functionalization. Overall, our results suggest that the AOP-based BN model can be used to provide a scientific basis for the development of safe nanomaterials.

Uncertainties and uncertain risks of emerging biotechnology applications: A social learning workshop for stakeholder communication.

Emerging applications of biotechnology such as new genomic techniques may give rise to new uncertainties and uncertain risks. Particularly the increased complexity and limited knowledge of possible risks associated with these new techniques, make it currently impossible to perform an adequate environmental risk assessment. As a result, there is a risk that such techniques don't get beyond experiments demonstrating the proof of principle, stifling their further development and implementation. To break free from this deadlock, we must be able to learn what such uncertainties and uncertain risks entail, and how they should be assessed to ensure safe further development. To shape a responsible learning environment to explore uncertainties and uncertain risks, we have organized five stakeholder workshops. By means of a case about the genetic engineering of plants' rhizosphere-an application abundant with uncertain risks-we identified tensions between different stakeholder groups and their different estimates of uncertainties and uncertain risks. Based upon derived insights, we developed a tool-a script for researchers to organize a stakeholder workshop-that enables a constructive discussion about emerging risks with a broad range of stakeholders. Thereby, the script provides a step-by-step approach to identify uncertainties, develop anticipatory strategies and adaptations in (experimental) research designs to lower or mitigate the earlier identified uncertainties, and helps to identify knowledge gaps for which (additional) risk research should be set up.

Identification of the safe(r) by design alternatives for nanosilver-enabled wound dressings.

The use of silver nanoparticles (NPs) in medical devices is constantly increasing due to their excellent antimicrobial properties. In wound dressings, Ag NPs are commonly added in large excess to exert a long-term and constant antimicrobial effect, provoking an instantaneous release of Ag ions during their use or the persistence of unused NPs in the wound dressing that can cause a release of Ag during the end-of-life of the product. For this reason, a Safe-by-Design procedure has been developed to reduce potential environmental risks while optimizing functionality and costs of wound dressings containing Ag NPs. The SbD procedure is based on ad-hoc criteria (e.g., mechanical strength, antibacterial effect, leaching of Ag from the product immersed in environmental media) and permits to identify the best one among five pre-market alternatives. A ranking of the SbD alternatives was obtained and the safer solution was selected based on the selected SbD criteria. The SbD framework was also applied to commercial wound dressings to compare the SbD alternatives with products already on the market. The iterative procedure permitted to exclude one of the alternatives (based on its low mechanical strength) and proved to be an effective approach that can be replicated to support the ranking, prioritisation, and selection of the most promising options early in the innovation process of nano-enabled medical devices as well as to encourage the production of medical devices safer for the environment.