Towards a reliable assessment of nanomaterial health effects using advanced biological models and assays

Det er et behov for å fremskaffe et godt vitenskapelig grunnlag for å beregne risiko for de som er eksponert til nanomaterialer (NMs), for å informere regulatoriske myndigheter samt å sikre en ansvarlig utvikling av nanoteknologi. Det er derfor et stort behov for å innhente pålitelig, balansert og objektiv informasjon og data for å redusere risikoen og usikkerheten rundt effekter av NMs. De fleste biologiske modeller og eksponeringssystemer, i tillegg til gjellende datamodeller, ikke representerer hva man eksponeres for i virkeligheten. Bruk av forsøksdyr har i mange tilfeller en begrenset kraft til å forutsi helseeffekter på mennesker. Dette krever et skifte fra kostbare og i mange tilfeller etisk tvilsomme dyreforsøk til innovative, pålitelige, kostnadseffektive og ikke minst sosialt aksepterte testmetoder og datamodeller. Formålet med NanoBioReal er å etablere virkelighetsnære metoder og biologiske modeller som gir en pålitelig, robust og effektiv plattform for å vurdere helseeffektene av NMs. Vårt testsystem dekker et stort område av biologiske modeller, fra enkeltceller til tredimensjonale (3D) cellemodeller som simulerer vev og organer. Dette inkluderer "air-liquid interface" (ALI) modeller for lungeeksponering, blod, og såkalte «organ-på-brikke» systemer (lunge- og mikrovaskulatur-på-brikke) som måler effekter i nåtid og fanger relevante effekter etter både kort- og langtidseksponeringer. For å unngå interferenser fra NMs, labelfri impedanse-baserte metoder ble etablert i prosjektet for å måle cytotoksistet og syklisk voltametri for oksidativ stress. Ett sett av representative NMs ble produsert og fysikalsk-kjemisk karakterisert. Deres cyto- og geno- toksisitet ble evaluert in vitro på 2D celle-modeller og resultatene sammenliknet med resultatene oppnådd ved bruk av avanserte 3D biologiske modeller og en dyremodell. Avanserte modeller for lunge, blodår og blod eksponering ble etablert og brukt i pågående toksistett, inflammasjon, oksidativ stress, og barriereeffekt testing. Et mikrofluidisk system for "label-free live" monitorering av celler og 3D biologiske modeller ble etablert og optimisering og øking av "throughput" ble gjort. NanoBioReal har levert pålitelige, robuste og relevante metoder, biologiske og data modeller for å støtte opp om «trygg ved design» tilnærmingen i utviklingen av NMs og for å svare på behovene til forskjellige interessenter og forvaltning. Nasjonale partnere: Inst. for klinisk odontologi (IKO), Det medisinske fak., Univ. i Bergen (UiB), Norsk instit. for luftforskning (NILU), Arbeidsmiljøinstituttet (STAMI), og Norges teknisk-naturvitenskapelige univ. (NTNU). Underleverandør: NorGenotech. Internasjonale partnere: Catalan Inst. of Nanoscience and Nanotechnology (ICN2), og Univ. i Gdansk. Samarbeidspartnere: Inst. for fysikk og teknologi (UiB), Inst. for elektrofag (HVL), og NIOM.

The NanoBioReal project contributes to a safer, more responsible and sustainable development of nanotechnology-based industry and use of its products by the society. This is based on addressing the urgent need to generate reliable science-based knowledge regarding the effects of nanomaterials (NMs) on human health and their safety in the working environment, as well as for consumers, and patients. The NanoBioReal project also addresses the RRI aspects outlined in the Nano2021 program. The project has lead to significant improvements of existing methods and biological models by getting closer to real-life exposure and thus enhancing their relevance, as well as by improving their efficiency through increased high throughput testing in order to keep up with the rapid pace of NM development. Furthermore, by maintaining current national nanosafety networks in Norway and establishing new networks with the European NanoSafety Cluster and the international network initiative on safe and sustainable nanotechnology (INISS), the consortium has had access to the best knowledge and expertise in Norway and in Europe. It consolidates the basis for the establishment of new networks, i.e., of a national “NanoSafety platform”. Close cross-cutting interaction of the partners within and between WPs, and their complementary skills has allowed the integration of their research and technology into a common innovative NanoBioReal approach. We have recruited and trained skilled professionals, such as the post-doc and young researchers to train competent staff with broad networks to the industry in the future. NanoBioReal also contributes with nanosafety aspects (UiB) in the Research School for Training the Next Generation of Micro- and Nanotechnology Researchers in Norway (TNNN). Moreover, NanoGenoTox (NGT) (SME participant in the project) will benefit from positioning the company as a reliable nanosafety assessment player. NGT has established new services suitable for NM safety assessment. The NanoBioReal results will contribute to helping other SME start-up companies to resolve HSE issues necessary for their sustainable growth. Overall, the industry will benefit as a result of the time- and cost-efficient, more relevant for the real situation and reliable hazard assessment platform developed in this project. This project contributes also to the improvement of environmental consequences, since similar biological effects could be relevant for organisms in the environment.

This proposal answers to the objective of the 5th thematic priority to expand the insight into the impacts of nanomaterials (NMs) on human health by addressing two of the main sources of uncertainty that hamper the assessment of real-life impact of NMs on human health: i) most of the existing in vitro and in silico models do not reflect real life exposure to NMs and ii) potential interferences of NMs with assays or detection systems. The aim of NanoBioReal is to provide solutions to these challenges by delivering i) beyond state of the art advanced biological models, from single-cells to organ-on-a-chip, representing target tissues and organs and ii) innovative realistic, reliable assessment tools integrated into an in vitro and in silico testing toolbox. The NMs selected for testing are relevant for Norwegian conditions and industry and include TiO2, ZnO, SiO2, nano-silver, nano-gold, and widely used polymer-based dental materials containing embedded nanoparticles. The key biological processes and mechanisms that will be investigated are: i) cellular uptake; ii) inflammation, iii) oxidative stress, iv) genotoxicity, DNA damage and DNA repair and v) cell death. At the end of the project a toolbox will be generated integrating reliable, robust, and efficient in-vivo relevant biological models and methods to support a safe-by-design approach to nanomaterial development and to answer the needs for nanomaterial hazard assessment of various end-users, stakeholders and regulators. The outcome of the project will contribute not only to bringing new mechanistic insights into the health impact of NMs, but also to reducing the uncertainty in health hazard and consequently in risk assessment, as well as to replacing animal use, in conformity with the 3R principles (reduce, refine, replace animal use).