Hazard assessment of nanoplastic particles mixed with bacteria

This project is a part of a large HORIZON 2020 EU, PlasticFate (Plastics fate and effects in the human body) that aims to improve our present understanding of the impact of micro- and nano-plastics (MP/NPs) and associated additives/adsorbed contaminants (A/C) in the human body. STAMI is leading the WP on Human hazard assessment.

Introduction: Human exposure to MP/NP may result from the widespread use of plastic products and their release to the environment, where they degrade to MP/NP particles. As consequence, these particles are found in food, drinking water, air and environmental media (food chain, soils). Despite recent efforts to assess the real dimension of human risks associated with MP/NP, our current knowledge is still insufficient. PlasticsFatE aims to address this challenge and associated uncertainties by implementing a comprehensive measurement and testing program ("test the test"), including inter-laboratory studies, to improve and validate the performance and applicability of available methods and tools to MP/NP.
 

The present MSc project will investigate the immunotoxic effects of MP/NPs contaminated by bacteria and bacterial constituents (lipopolysaccharide and lipoteichoic acid). We hypothesized that contamination of nanoplastics by Gram- or Gram+ bacteria may have significant impacts on their immunotoxic effects particularly on neutrophils in the lungs. We will therefore study the initial triggering mechanisms, i.e. cell death (apoptosis and netosis), oxidative stress and inflammation in neutrophil-like cells differentiated from HL60. If possible, we will also study some key cell signalling pathways ( eg TLRs, NF-kB) implicated in modulating allergic and non-allergic inflammation.

Materials and methods: HL60 cells differentiated into neutrophil-like will be exposed to specific nanoplastic particles mixed with Pseudomonas sp or Bacillus subtilis. Adverse outcomes will be assessed using various approach including luminex technology, gene expression, western blot analysis, microscopic and flow cytometric analysis. In addition, initial characterization of the mixed plastic materials and microorganisms using FESEM (Field emission Scanning Electron microscope) will be performed.

The experimental work will include the following tasks:

Characterization of exposure material using FESEM (Field Emission Scanning Electron Microscope)

Cell culture techniques: maintenance, seeding, differentiation and in vitro exposure

                Bioassays for cell death (apoptosis and Netosis), oxidative stress and cytokines

                RNA expression by qPCR

                Reporter cell assays

Contact:
Anani Komlavi Afanou

Forsker/Associate Research Professor

Arbeidstoksikologi gruppen

Statens arbeidsmilj?institutt (STAMI)

National Institute of Occupational Health

+47 23195247 / +47 95080052

E-post: anani.afanou@stami.no

www.stami.no

Bes?ksadresse: Gydas vei 8, Majorstuen, Oslo

Postadresse: Pb 5330 Majorstuen, 0304 Oslo, Norway