Partners background

NILU – Norsk Institutt for Luftforskning

GLP systemet og alle medfølgende SOP’er, inkludert beskrivelse av testmetoder. Alle resultater som er oppnådd i tidligere prosjekter,
som f.eks testmetoder og teknikker som er utviklet spesielt med tanke på testing av nanomateriale, dispersjonsprotokoller og
protokoller for karakterisering (f.eks Nanosight eller TEM).

SINTEF

Erfaring og Standard Operating Procedures (SOPs), inkludert beskrivelse av testmetoder som er etablert ved deltakelse
i EU prosjektet NANoREG vil bli brukt i dette prosjektet. Alle resultater oppnådd i tidligere prosjekter, slik som test metoder
og teknikker utviklet spesielt for vurdering av nanomaterialer, dispersjonsprotokoller og protokoller for karakterisering av nanomaterialer.
Bakgrunnsmateriale slik som celler involvert i ”high throughput sceening” metode og nanomaterialer kan, etter tillatelse av våre eksterne
partnere og etter avtale med SINTEF, bli delt med prosjektdeltarkere i NorNANoREG. Tilsvarende vil metoder som er etablert for
undersøkelse av nanomaterialers skjebne og effekter i miljø, i dette prosjektet bli delt med prosjektpartnere i NorNANoREG.           

Universitetet i Bergen 

Resultater oppnådd i FP7 «NANoREG» prosjektet der UiB deltar i WP5, særlig task 5.6, der inntil nå grunnarbeidet ble gjort for å etablere
SOPs for dispersjon av nanomaterialer brukt i NANoREG (noen av disse skal også brukes i NorNAnoREG), samt etablering av metoder for
nano-cytotoksisitet.  Hovedfokuset er på high-throughput metoder: impedanse-basert (i samarbeid med Emil Cimpan, Elektronikk og automasjon,
HiB, Bergen og Uwe Pliquett, IBA, Heiligenstadt, Tyskland), PCR super-array, microarray (i samarbeid med Kamal Mustafa, Ying Xue,
Julia Schoelermann, IKO, UiB, Bergen), micro / nano-fluidics platform (i samarbeid med Bodil Holst, Nanofysikk – IFT, UiB, Julia Schoelermann,
IKO, UiB, og Jan Kocbach CMR instrumentation, Bergen og Raghavendra Palankar og Mihaela Delcea, Nanostructures lab., Univ. of Greifswald, Tyskland),
Ultrahigh Resolution Imaging (samarbeid med Nils Roar Gjerdet, IKO, UiB), TIRF, TEM og SEM mikroskopi (i samarbeid med Julia Schoelermann). 

Relevante publiksjoner:

1.   “An impedance-based high-throughput method for evaluating the cytotoxicity of nanoparticles”. E Cimpan, T Mordal, J Schoelermann, U Pliquett, MR Cimpan. J of Physics: Conference Series. 429. 012026 doi:10.1088/1742-6596/429/1/012026, 2013;
2.    “Role of physico-chemical characteristics in the uptake of TiO2 nanoparticles by fibroblasts” Z Allouni, PJ Høl, MA Cauqui, NR Gjerdet, and MR Cimpan – Toxicol in vitro 26(3):469-79, 2012; “How dangerous is nanotechnology: nanotoxicology an interdisiciplinary field” MR Cimpan, NR Gjerdet, Naturen 3: 25-2, 2012;
3.    “Agglomeration and Sedimentation of TiO2 Nanoparticles in Cell Culture Medium” ZE Allouni, MR Cimpan, PJ Høl, T Skodvin, NR Gjerdet. Colloids and Surfaces B: Biointerfaces 68(1):83-87, 2009
4.   “Induction of cell death by TiO2 nanoparticles: studies on a human monoblastoid cell line” CI Vamanu, MR Cimpan, PJ Høl, S Sørnes, SA Lie, NR Gjerdet. Toxicol in Vitro 22(7): 1689-1696, 2008;

Submitted:

1. “Contact-dependent transfer of TiO2 nanoparticles between mammalian cells”. J Schoelermann, ZE Allouni, A Burtey, HH Gerdes, MR Cimpan;
2. “The effect of blood protein adsorption on cellular uptake of TiO2 nanoparticles” ZE Allouni, PJ Høl, MA Cauqui, NR Gjerdet, MR Cimpan;

Abstracts:
“A microfluidic chip-based impedance flow cytometry method for evaluating the cytotoxicity of nanoparticles” M Nerland, M Di Berardino, G Schade, E Cimpan, K Mustafa, MR Cimpan. NANOTOX 2014, Antalya, Turkey;
 “Role of particle size on uptake of nano-TiO2 by human osteoblasts” M Ibrahim, M Nerland, K Mustafa, MR Cimpan. NANOTOX 2014, Antalya, Turkey;

Norges Miljø- og biovitenskapelige universitet (NMBU)

Experience ​​and Standard Operating Procedures (SOPs), including description of test methods, established through participation in the EU project NANoREG will be used in this project. All results obtained in previous projects, such as test methods and techniques developed specifically for assessment of nanomaterials and protocols for nanomaterial characterisation (e.g. electron microscopy, DLS). Background material such as radiolabelling techniques, synchrotron methods and ultrafiltration can, after permission by our external partners and agreement with NMBU, be shared with project partners in NorNANoREG. Similarly, methods that are established on Environmental fate and effects in this project will be shared with project partners in NorNANoREG

Statens Arbeidsmiljøinstitutt

Erfaringer gjort og SOPs etablert gjennom deltakelse i EU-prosjektet NanoReg vil bli benyttet i dette prosjektet. Bakgrunnsmaterialet slik som celler som inngår i cellemodellen og nanomaterialer kan etter tillatelse fra våre eksterne samarbeidspartenere og avtale med STAMI deles med prosjekt partnere i NorNanoReg. Likeledes kan metoder som blir etablert på STAMI i dette prosjektet deles med prosjektpartnere i NorNanoReg.

Genøk – Senter for biosikkerhet

Previous work on the theory and practice of risk governance, transdisciplinary research, and responsible research and innovation (RRI) will be used as project background. The approaches to safety by design and safe innovation as developed thorugh the EU NANoREG project will also be shared with partners of the NorNanoReg project and used as appropriate.

Comet Biotech AS

Experience with high throughput methods for measuring DNA damage (based on methods developed in the EC FP6 project ‘COMICS’) will comprise background knowledge, together with protocols for various modified versions of the assay, such as to measure oxidised DNA bases. The DNA repair enzyme formamidopyrimidine glycosylase will be available. Reference: Azqueta,A., Collins,A.R. (2013) The essential comet assay: a comprehensive guide to measuring DNA damage and repair. Arch. Toxicol. 87, 949-968

 

Project structure

WP1 tasks and deliverables

Tasks:
Physical and chemical characterisation of MNMs provided by NANoREG (NMBU, SINTEF)
Implementation of MNM dispersion SOPs for environmental exposure media (SINTEF, NMBU)
Characterisation of MNMs in exposure media and organisms (NMBU, SINTEF)

Deliverables:
Validated SOP for obtaining reproducible and stable MNMs in aquatic (ecotoxicology) media.
SOPs for in vitro and in vivo toxicity studies (collaboration with WP2 and WP3).
Articles in peer reviewed literature (3-4 papers).

WP2 tasks and deliverables

Tasks: (All tasks go beyond NANoREG).
Validation of SOPs for cellular uptake (URI), proliferation, and cell death (impedance-based methods) (UiB), cytotoxicity, genotoxicity (NILU, CBT), mutagenicity (NILU); Bhas 42 CTA (NILU), the human lung transformation assay in vitro (hLCTA) (STAMI).
Validation of hLCTA for carcinogenic potential of MNMs by assessing tumorigenicity of transformed cells in nude mice. (Beyond the brief of the NANoREG project) (STAMI).
Setting up an ALI-exposure monoculture and co-culture model (NILU)
Development of novel HTS methods for evaluating uptake and intercellular transfer of MNM (UiB), genetic and epigenetic changes (NILU).
Development of micro-fluidic systems for high-throughput cell-based assays, which provide a quantitative spatial and temporal control of MNM dose and interaction time, and mimick in vivo physiological environment (UiB).
Identification of new sensitive molecular biomarkers by investigating perturbations (genetic, epigenetic and transcriptomic) in relevant biological pathways. (NILU, UiB, CBT, STAMI)

Deliverables:
Comparison, evaluation, and validation of proposed in vitro methods for assessing nanotoxicity.
Comparison of in vitro effects with published in vivo data from inhalation and ALI studies .
Identification of the most sensitive molecular biomarkers of carcinogenicity and tumorigenicity.
Ranking of MNMs according to cytotoxicity, genotoxicity, mutagenicity, transformational/tumorigenic ability; identification of physico-chemical characteristic responsible for toxicity.
SOPs for HTP methods for human-related in vitro toxicity assessment.
Publications in peer reviewed literature (5-7 papers).

WP 3 tasks and deliverables

Tasks:
Validation of aquatic and terrestrial ecotoxicity SOPs selected in NANoREG for determining acute toxicological responses at the cellular and individual levels (SINTEF, NMBU).
Determination of bioavailability, internal exposure and uptake of MNMs to aquatic and terrestrial organisms (NMBU, SINTEF).
Development and standardisation of HTP ecotoxicity approaches for algae, bacteria and C.elegans (SINTEF, NMBU).

Deliverables:
OECD aquatic and terrestrial toxicity levels, EC50/PNEC-values for different MNMs, ranking of toxicity of examined MNMs and within MNM classes (e.g. CNTs).
Bioaccumulation factors for examined MNMs.
Suggested SOPs for HTP ecotoxicity assessment.
Publications in peer reviewed literature (3-4 papers).

WP 4 Tasks and deliverables

Tasks:
Experiment with innovative methods of interdisciplinary communication to encourage information exchange and learning across the project partners and WPs at each of the annual project meetings
Facilitate researchers to anticipate potential challenges and identify available alternatives in their work and to reflect on important uncertainties, underlying values, and embedded assumptions
Explore potential for enhanced interaction and communication among toxicologists, technology developers, regulators and other relevant stakeholders to ensure mutual learning, dissemination of results and implementation of new or modified test strategies.

Deliverables:
Policy brief on challenges on the horizon for ensuring the health and safety of MNMs
Academic article on important uncertainties, assumptions and choices shaping our knowledge of the different impacts of MNMs on human health and ecosystems
Workshop with a range of relevant stakeholders to discuss the project’s approach and results
Proposed guidelines for regulatory bodies on robust and reliable MNM testing strategies

WP5 tasks

Yearly partner meetings will be organized to monitor progress and to plan joint activities, discuss results and cross cutting topics, prepare reports, common publications, etc. Altogether, 3 project meetings are planned, and in addition an open workshop with various stakeholders.
To promote faster transfer of knowledge and career development of young scientists, exchange of postdocs will be encouraged, e.g. between NILU/CometBiotech/ UiB/STAMI and NMBU and SINTEF.



WP participation
    

 Partner name

 WP1

WP2

WP3

WP4

WP5

 NILU

 

 X

 X

 SINTEF

 X

 

 X

 X

 

 UiB

 

 

 X

 

 STAMI

 

 X

 

 X

 

 UMB

 

 X

 

 GenØk

 

 

 

 X

 X

 CBT

 

 X

 X

 X

The Need for Reliable Testing of Nanomaterial Risks

Possible negative impacts on human health and the environment from increasing exposure to manufactured nanomaterials (MNMs) is a growing concern. As products incorporating nanomaterials are continually being released onto the market, there is now an urgent need to create a sound scientific basis from which to assess potential risks to workers, consumers and ecosystems. There are, however, currently no commonly agreed upon methods for generating robust and reliable knowledge about the health and environmental risks posed by novel nanomaterials; a situation that threatens to hamper both the innovative potential of nanomaterials and their public acceptance. To understand the potential risks to human health and the environment, it is urgent that reliable methods and common standards for regulatory testing are developed and applied as soon as possible.

NorNanoReg: A national initiative to develop a common approach to regulatory testing
NorNanoReg is an NFR funded national initiative towards the development of a common approach to the regulatory testing of manufactured nanomaterials (MNMs).

The goals of the NorNanoReg project are to:
1. Evaluate the adequacy and appropriateness of using existing methods for both characterizing MNMs and assessing their health and safety impacts
2. Develop novel models and methods for testing where necessary, particularly emphasizing high throughput techniques to allow rapid testing of the large and growing number of MNMs.
3. Generate relevant information on the potential adverse health and environmental impacts of MNMs using reliable methods
4. Encourage the research results to inform risk assessment and regulatory decision-making on MNMs
5. Advance effective transdisciplinary communication between those developing MNMs, those studying their risks, and those responsible for their regulation.

NorNanoReg is coordinated by the Norwegian Institute for Air Research (NILU) and involves a further six national partner institutes: the University of Bergen, the Norwegian University of Life Sciences, SINTEF, the National Institute of Occupational Health (STAMI), GenØk Centre for Biosafety, and Comet Biotech.

NANoREG: A European initiative to develop a common approach to regulatory testing
NorNanoReg represents the Norwegian national initiative and consortium within a broader European effort to develop a common approach to the regulatory testing of nanomaterials. NANoREG (www.nanoreg.eu) is an FP7 flagship project involving 16 participating countries and 63 partner institutes. Based on questions and requirements supplied by regulators and legislators, NANoREG seeks to: (i) provide answers and solutions from existing data, complemented with new knowledge, (ii) provide a tool box of relevant instruments for risk assessment, characterisation, toxicity testing and exposure measurements of MNMs, (iii) develop, for the long term, new testing strategies adapted to innovation requirements, (iv) establish a close collaboration among authorities, industry and science leading to efficient and practically applicable risk management approaches for MNMs and products containing MNMs.

Contact & further information
For further information about the project and its results, you can contact:
Lise Fjellsbø, NILU, e-mail: lbf@nilu.no