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.
Backgroung: 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 fungi and fungal constituents (mycotoxins and beta-glucans). We hypothesized that contamination of nanoplastics by fungal spores or mycelial fragments 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 the genotoxicity and 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 fungal particles from Aspergillus fumigatus. 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
Autumn Aug-Sept-Oct 2021 |
Training in cell-, Microbio- and Nanolab techniques Preparation and characterization of the exposure materials Work with cell culture and HL60 exposure |
Autumn Nov-Des 2021 |
Reporter cell assays |
Spring 2022 Jan-Mars 2022 |
Bioassays RNA isolation |
Spring 2022 April-Jun 2022 |
qPCR and data analysis |
Autumn 2022 Aug-Oct 2022 |
Finish lab work Analysis and thesis writing from Oct 2022 |
Expected final exam |
Juni 2023 |
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