Projects / Programmes source: ARIS

Elaboration and evaluation of a toxicity test for engineered nanoparticles with terrestrial isopods

Research activity

Code Science Field Subfield
1.03.01  Natural sciences and mathematics  Biology  Zoology and zoophysiology 

Code Science Field
B361  Biomedical sciences  Physiology of invertebrates 
B130  Biomedical sciences  Physiological biophysics 
B740  Biomedical sciences  Pharmacological sciences, pharmacognosy, pharmacy, toxicology 
toxicity testing, nanoparticles, biomarkers, invertebrates, electron microscopy
Evaluation (rules)
source: COBISS
Researchers (18)
no. Code Name and surname Research area Role Period No. of publicationsNo. of publications
1.  22304  PhD Mojca Bavcon Kralj  Chemistry  Junior researcher  2007  200 
2.  11155  PhD Damjana Drobne  Biology  Head  2007 - 2009  859 
3.  11215  PhD Samo Drobne  Geodesy  Researcher  2007 - 2009  875 
4.  10582  PhD Ingrid Falnoga  Neurobiology  Researcher  2007 - 2009  343 
5.  05935  PhD Nina Gunde-Cimerman  Biotechnology  Researcher  2007 - 2009  1,255 
6.  18749  PhD Rok Kostanjšek  Biology  Researcher  2007 - 2009  469 
7.  21408  PhD Aleš Lapanje  Microbiology and immunology  Researcher  2007  354 
8.  11873  PhD Urška Lavrenčič Štangar  Chemistry  Researcher  2007 - 2009  554 
9.  24292  PhD Vladka Lešer  Public health (occupational safety)  Junior researcher  2007 - 2009  157 
10.  08259  PhD Domen Leštan  Plant production  Researcher  2007 - 2009  407 
11.  21387  PhD Branka Mozetič Vodopivec  Plant production  Researcher  2007  142 
12.  26053  PhD Živa Pipan Tkalec  Biology  Junior researcher  2007 - 2009  42 
13.  07560  PhD Maja Remškar  Physics  Researcher  2007 - 2009  829 
14.  11539  PhD Polonca Trebše  Control and care of the environment  Researcher  2007 - 2009  510 
15.  18846  Barbara Trobec    Technical associate  2007 - 2009  45 
16.  08437  PhD Goran Turk  Civil engineering  Researcher  2007 - 2009  524 
17.  27543  PhD Janez Valant  Pharmacy  Junior researcher  2007 - 2009  56 
18.  16065  PhD Primož Zidar  Biology  Researcher  2007 - 2009  171 
Organisations (4)
no. Code Research organisation City Registration number No. of publicationsNo. of publications
1.  0106  Jožef Stefan Institute  Ljubljana  5051606000  90,038 
2.  0481  University of Ljubljana, Biotechnical Faculty  Ljubljana  1626914  66,599 
3.  0792  University of Ljubljana, Faculty of Civil and Geodetic Engineering  Ljubljana  1626981  26,383 
4.  1540  University of Nova Gorica  Nova Gorica  5920884000  13,886 
There are risk issues with every new technology. With the increasing presence of nanomaterials in commercial products, a growing debate is emerging on whether the environmental and social costs of nanotechnology outweigh its many benefits. Information about safety and potential hazards of nano-sized particles is urgently needed. To date, only a few studies have investigated the toxicological effects of nanomaterials and no clear guidelines exist on how to quantify these effects. In the proposed work we will modify the existing test for soluble chemicals to be applicable for nanomaterials. We plan to develop relevant, robust, reproducible and relatively cheap toxicity tests for engineered nanomaterials with the terrestrial isopod Porcellio scaber (Isopoda, Crustacea). Terrestrial isopods have been well characterised for their relevance in (eco)toxicity studies. In toxicity tests with terrestrial isopods, biomarkers on various levels of biological organization are assessed, and values for effect concentration (EC) or lowest observed effect concentration (LOEC), no observed effect concentration (NOEC) values as well as effect level (EL), lowest observed effect level (LOEL) OR no observed effect level (NOEL) for tested chemicals are generated. We aim to produce a toxicity test that: a) Can be used as a screening tool before other laboratory tests (relevant to humans or environment) are selected; b) Can be used in comparative toxicity studies of nanoparticles and their corresponding bulk material; c) Can be used as a toxicity test of choice for environmental samples. Conventional transmission electron microscopy (TEM) and focused ion beam (FIB)/scanning electron microscopy (SEM) will be used to characterise tested nanoparticles. Intracellular distribution of nanoparticles will also be investigated by TEM and FIB/SEM. Results of the project will be published in scientific papers and shared in the Nano Safety Reporting System. With our findings we will participate in risk communication with public at both national and international levels. We believe that the concerns about nanoparticles’ toxicity must be addressed while the field is still young and exposures limited.
Significance for science
The aim of the project was to investigate time- and dose-dependent in vivo effect of ingested nanosized TiO2, nanoZnO, nanoAg and nanoCu. Analysis of the effects of nanoparticles on cell membrane stability, lipid peroxidation and mitochondrial membrane potential on digestive gland cells of the model organism (Porcellio scaber, Isopoda, Crustacea) under realistic in vivo conditions allows identification of interactions between the epithelium of the digestive gland and nanoparticles. 1) The results suggest that ingested nanosized TiO2 affects first the digestive gland cell membrane. Prolonged exposure and higher exposure concentrations caused also intracellular perturbations including oxidative stress. The three measured cytotoxicity responses showed different patterns of dose and time response. Cell membranes are affected by nanosized TiO2 before oxidative stress or metabolic changes are recorded. We observed a threshold response for membrane destabilization. Lipid peroxidation was time-dependent and mitochondrial function was neither time nor dose dependent. Oxidative stress as measured by lipid peroxidation was accompanied by biogenesis of mitochondria and cell membrane destabilization. The in vivo experimental conditions used allow assessment of conventional cytotoxicity biomarkers as a function of exposure to nanoparticles. 2) In case of nanoZnO, nanoAg and nanoCu similar effects were observed, but at the present state of knowledge it is not possible to distinguish the effect of nanoaprticles from the effct of disoluted ions from nanoparticles. For example in case of nanoZnO, nanoCu and nanoAg, dissolution is well documented and the major “nano” effect is actually attributed to these ions and not nanoparticles. 3) The major difference in testing the effects of nanoparticles and soluble chemicals is that nanoparticles must be well characterised. Both, primary characteristics (particle size, crystal structure etc.) must be known as well as secondary characteristics, which depend on the medium, where nanoparticles are suspended (size of aggregates, zeta potential etc. 4) Oxidative stress is involved in a cascade of effects cased by nanoparticles but it is not the initial response. Most probably nanoparticles interact first with cell membrane and subsequently with higher concentration or prolonged exposure other cellular effects occur. 5) A model test organism Porcellio scaber (Isopoda, Crustacea) proved to be well suited to assess biological potential of different nanoparticles. In our project we succeeded to elaborate a battery of biomarkers, which could substantially help in environmental risk assessment (ERA). The results of our project are most beneficial in environmental hazard characterization and in comparative toxicity studies of chemical pollutants and products of new technologies also in Slovenia.
Significance for the country
To date, few studies have investigated the toxicological and environmental effects of direct and indirect exposure to nanomaterials and no clear guidelines exist to quantify these effects. One reason for that lies in the fact that it is not clear whether the existing methods for toxicity testing are useful also for nanoparticles. The results of our study answered part of this question and provided a useful tool for nanoparticle risk characterisation that could be used for variety of nanopartilcles. In our project we succeeded to elaborate a battery of biomarkers, which could substantially help in hazard characterisation of nanoparticles and environmental risk assessment (ERA). The results of our project are most beneficial in environmental hazard characterization and in comparative toxicity studies of chemical pollutants and products of new technologies also in Slovenia.
Most important scientific results Annual report 2008, final report, complete report on dLib.si
Most important socioeconomically and culturally relevant results Annual report 2008, final report, complete report on dLib.si
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