“Need for new biological assays for new nanomaterials”
1.5-day conference organized by NanoTOES FP7 project
10th and 11th July 2012, C006, Health Sciences Centre, University College Dublin, Belfield, Dublin 4
Conference Programme & Abstract book
About NanoTOES NanoTOES: Training of Experts in NanoSafety, an FP7 Marie Curie Initial Training Network Grant number: 264506 Coordinator: Paris Lodron University of Salzburg, Prof. Albert Duschl, [email protected] www.nanotoes.eu NanoTOES 1st International Conference 1
Scientific Committee
Albert Duschl, Paris Lodron University of Salzburg
Diana Boraschi, Institute of Biomedical Technologies
Maria Dusinska, Norwegian Institute for Air Research
Yvonne Kohl, Fraunhofer Institute for Biomedical Engineering IBMT
Iseult Lynch, University College Dublin
Emilia Madarasz, Institute of Experimental Medicine
Markus Pesch, Grimm Aerosol Gmbh
Victor Puntes, The Catalan Institute of Nanotechnology
Michael Riediker, Institute for Work and Health
Claus Svensden, Centre for Ecology and Hydrology
Matthias Voetz, Bayer Technology Services GmbH
Colin Wilde, AvantiCell Science Ltd
Local Organising Committee
Iseult Lynch, University College Dublin
Louise McCarron, University College Dublin
Bogumila Reidy, University College Dublin
Elisabeth Eppacher, Paris Lodron University of Salzburg
NanoTOES 1st International Conference 2
Need for new biological assays for new nanomaterials
Day 1 – Tuesday 11th July 2012
Location: Lecture Theatre C006, Health Sciences Centre, University College Dublin
13.00 Registration & Lunch
14.30 Opening of conference – Albert Duschl & Iseult Lynch
Session 1 – Beyond cytotoxicity – new assays to account for complexities of Nanomaterials (Chair: Hagen Briesen)
14.35 – 15.10 Maria Costantini (Health Effects Institute) – Health effects of ultrafine particles: lessons from air pollution science
15.10 – 15.45 Andrea Haase (BfR) - Effects of silver nanoparticles on primary mixed neural cell cultures: Uptake, oxidative stress and acute calcium responses
15.45 – 16.15 Coffee
16.15 – 16.50 Jürgen Palluhn (Bayer Health) - Subchronic inhalation toxicity of iron oxide particles in rats: pulmonary toxicity is determined by the particle kinetics typical of poorly soluble particles.
16.50 - 17.10 Emilia Izak (Bayer) - Interaction of differently functionalized silica NPs with neural stem- and tissue-type cells (NanoTOES ESR)
17.10 – 17.30 Anna Huk (NILU) – Is toxic potential of nanosilver dependent on its size? Possible mechanisms of toxicity. (NanoTOES ESR)
17.30 – 18.00 General discussion
19.30 Conference dinner
Ely Bar & Brasserie IFSC, Dublin 1
NanoTOES 1st International Conference 3
Need for new biological assays for new nanomaterials
Day 2 – Wednesday 11th July 2012
Location: Lecture Theatre C006, Health Sciences Centre, University College Dublin
09.00 – 09.05 Opening Day 2 – Albert Duschl
Session 2 – Biological assays adapted to nanomaterials surface properties (Chair: Diana Boraschi)
09.05 – 09.40 Jason Unrine (University of Kentucky) – Environmental transformations of silver nanoparticles: the role of particle coatings
09.40 – 10.15 Kenneth Dawson (University College Dublin) – Role of protein corona in uptake, localization and (delayed) impacts of nanomaterials
10.15 – 10.35 Mun Li Yam (AvantiCell) - Standardised assay platforms for cell-based nanosafety testing (NanoTOES ER)
10.35 – 10.55 Carolin Schultz (NERC) - Media effects on silver nanoparticle bioavailability and toxicity in Caenorhabditis elegans (NanoTOES ESR)
10.55 – 11.25 Coffee
11.25 – 12.00 Olga Tsyusko (University of Kentucky) - Toxicity and molecular level effects of gold and silver nanoparticles on soil invertebrates, C. elegans and Eisenia fetida
12.00 – 12.30 General discussion
12.30 – 14.30 Lunch and poster session
Session 3 – Special considerations for immunosafety (Chair: Maria Dusinska)
14.30 – 15.05 Yvonne Kohl (Fraunhofer Institute for Biomedical Engineering) - Biocompatible micro-sized cell culture chamber for detection of NP- induced IL8 promoter activity on a small cell population.
15.05 – 15.40 Margriet Park (RIVM) - The effect of particle size & biointeractions on the inflammation, developmental toxicity and genotoxicity of silver NPs.
15.40 – 16.05 Coffee
16.05 – 16.40 Diana Boraschi (CNR) - Nano-immunosafety: issues in assay validation 16.40 – 17.10 General Discussion
17.10-17.15 Final remarks and close of meeting NanoTOES 1st International Conference 4
Presentation Abstracts
NanoTOES 1st International Conference 5
Health Effects of Ultrafine Particles: Lessons from Air Pollution Science Maria Costantini Health Effects Institute, 101 Federal Street, Boston, MA 02110, USA [email protected]
Ambient particulate matter (PM) is a complex mixture of particles suspended in the air that vary in size and composition depending on their sources. Particles emitted from mobile sources tend to fall into a bimodal distribution referred to as nuclei mode and accumulation mode depending on the mechanism of formation1. Nuclei mode particles are less than 50 nm in diameter and are generally made of hydrocarbons, sulfur, or metallic ashes. Accumulation mode particles range in size from about 50 nm to 500 nm and may contain elemental and organic carbon, nitrate, sulfate, and various metallic ashes.
Ambient particles are generally categorized into three size groups: ultrafine (< 100 nm), fine (between 1,000 and 2,500 nm), and coarse (>2,500 nm and up to 10,000 nm). Ultrafine PM includes nuclei mode and a fraction of accumulation mode PM.
The US Environmental Protection Agency and other agencies around the world regulate the mass concentration of ambient particles smaller than 10,000 nm in diameter (PM10). Some agencies, including the US EPA, also regulate the mass concentration of particles smaller than 2,500 nm in diameter (PM2.5). Ambient concentrations of ultrafine particles are not currently regulated. While these particles contribute very little to the mass of PM2.5 and PM10, they are present in high numbers. Their level in ambient air is generally reported as number count per cubic cm of air. Ultrafine PM consists of a carbon core with absorbed organic compounds, inorganic ions, and metallic ash. Dosimetry information for ultrafine ambient PM relies on deposition models for solid particles that are relevant also to engineered nanoparticles.
Experimental studies in rodents and humans to determine the role of particle size and composition have been conducted using laboratory-generated ultrafine and fine PM made up of one element (for example carbon, titanium, iron, or nickel). These “model particles” are used by both the air pollution and the nanotechnology health assessors. Studies with ultrafine PM sampled from ambient air have provided complementary data to help validate the results with model particles. By looking at the emerging picture of deposition and mechanistic information, pathways by which ultrafine particles may cause effects are beginning to emerge (see as examples 2, 3, 4). What we have learned so far is that inhaled ultrafine particles can cause a variety of effects that depend on their physico-chemical properties and the characteristics of the host. Composition and solubility, and perhaps physical parameters such as surface area or surface charge, are important characteristics. Some ultrafine particles may cause airway inflammation (i.e., increased production of reactive oxygen species and increased number of inflammatory cells.) Others have been shown to cause effects on endothelial cells lining the blood vessels, blood coagulation, and cardiac function.
This presentation will summarize, and provide examples, of what we have learned about dose metrics and mechanisms of action from studies of ultrafine ambient particles and model NanoTOES 1st International Conference 6
particles and discuss how this information can be helpful for studies of engineered nanoparticles.
References 1. D.B. Kittelson, W.F.Watts, J.P. Johnson, Atmos. Environ., 2004, 38, 9-19. 2. J.A. Araujo, B. Barajas, M. Kleinman, X. Wang, B.J. Bennett, K.W. Gong, M. Navab, J. Harkema, C. Sioutas, A.J. Lusus, A.E. Nel, Circul. Res., 2008, 102, 589-596. 3. W.E. Cascio, E. Cozzi E, S. Hazarika, R.B. Devlin, R.A. Henriksen, R.M. Lust, M.R van Scott, C. J. Wingard, Inhal. Toxicol., 2007, 19 Suppl 1, 67-73. 4. G. Oberdorster, E. Oberdorster, J. Oberdorster, Environ Health Perspect, 2005,113, 823- 839.
NanoTOES 1st International Conference 7
Effects of silver nanoparticles on primary mixed neural cell cultures: Uptake, oxidative stress and acute calcium responses
Andrea Haase1, Stephanie Rott2, Alexandre Mantion3, Philipp Graf4, Andreas F. Thünemann3, Wolfgang P. Meier4, Andreas Taubert5, Andreas Luch1, and Georg Reiser2
1) German Federal Institute for Risk Assessment (BfR), Department of Product Safety, Max- Dohrn- Strasse 8- 10, 10589 Berlin, Germany, 2) University of Magdeburg, Institute of Neurobiochemistry, Leipziger Strasse 44, 39120 Magdeburg, Germany, 3) BAM — Federal Institute for Materials Research and Testing, Unter den Eichen 87, 12205 Berlin, Germany, 4) University of Basel, Department of Chemistry, Klingelbergstrasse 80, 4056 Basel, Switzerland, 5) University of Potsdam, Institute of Chemistry, Karl-Liebknecht-Strasse 24-25, 14476 Potsdam- Golm, Germany, [email protected]
In the body, nanoparticles can be systemically distributed and then may affect secondary target organs such as the central nervous system (CNS). Putative adverse effects on the CNS are rarely investigated to date. Here we used a mixed primary cell model consisting mainly of neurons and astrocytes and a minor proportion of oligodendrocytes to analyze the effects of well-characterized 20 nm and 40 nm silver nanoparticles (SNP). Similar gold nanoparticles served as control and proved inert for all endpoints tested. SNP induced a strong size-dependent cytotoxicity, which was also dependent on the differentiation state of the cells in the low concentration range (up to 10 µg/ml of SNP). For detailed studies, we used low/medium dose concentrations (up to 20 µg/ml) and found strong oxidative stress responses. Reactive oxygen species (ROS) were detected along with formation of protein carbonyls and the induction of heme oxygenase-1. We observed an acute calcium response, which clearly preceded oxidative stress responses. ROS formation was reduced by antioxidants, whereas the calcium response could not be alleviated by antioxidants. Finally, we looked into the responses of neurons and astrocytes separately. Astrocytes were much more vulnerable to SNP treatment compared to neurons. Consistently, SNP were mainly taken up by astrocytes and not by neurons. Immunofluorescence studies of mixed cell cultures indicated stronger effects on astrocyte morphology. Altogether, we can demonstrate strong effects of SNP associated with calcium dysregulation and ROS formation in primary neural cells, which were detectable already at moderate dosages.
References
Haase A., Rott S., Mantion A., Graf P., Plendl J., Thünemann A.F., Meier W.P., Taubert A., Luch A., Reiser G. Effects of silver nanoparticles on primary mixed neural cell cultures: Uptake, oxidative stress and acute calcium responses. Toxicol. Sci. 2012, 126, 457- 68.
NanoTOES 1st International Conference 8
Subchronic inhalation toxicity of iron oxide particles in rats: pulmonary toxicity is determined by particle kinetics typical of poorly soluble particles.
Jürgen Pauluhn Bayer HealthCare Pharma Research Center, Aprather Weg, 42096 Wuppertal, Germany [email protected]
Two competing mode of actions have to be envisaged when examining the inhalation toxicity of poorly soluble iron oxides. One is related to surface chemistry, dissolution, and liberation of redox-active free ferric/ferrous iron, the other has to account for particle accumulation within the pool of alveolar macrophages and at what stage of bioaccumulation an increased exhaustion of the phagocytic capacity occurs1,2. This most critical hallmark of kinetic lung overload needs to be robustly charac- terized to better understand whether lung toxicity is caused by particokinetics (accumulation) or substance-specific
particodynamics. Fe3O4 particles retained in the lung display kinetic features typical of poorly soluble particles3,4 (Fig. 1). Aggravation of lung toxicity due to free iron was not apparent, even under conditions of kinetic lung overload.
Figure 1. Cornerstones of Fe3O4 particokinetics in rats
Collectively, inhalation bioassays with poorly soluble particles (PM) follow the paradigm that the total accumulated dose of PM determines the magnitude pulmonary toxicity. Likewise, reversibility is clearly dependent on particokinetics and the total cumulative dose delivered to the lung (Fig. 1). The no-effect levels (NOAELs) with Fe3O4 from 4- and 13-week rat repeated exposure inhalation studies matched those predicted by kinetic modeling. Hence, mechanism-based mathematical modeling proved to be highly supportive for designing new inhalation bioassays as well as for estimating safe chronic occupational exposure levels.
References
1. J. Pauluhn, Inhal. Toxicol., 2009, 21(S1), 40-54. 2. J. Pauluhn, Toxicology, 2011, 279, 176-188. 3. J. Pauluhn, J. Appl. Toxicol., 2012, 32, 488-504. 4. J. Pauluhn and W. Wiemann, Inhal. Toxicol., 2011, 23, 763-783.
NanoTOES 1st International Conference 9
Interaction of differently functionalized fluorescent silica NPs with neural stem- and tissue-type cells.
E. Izak1, 3, K. Kenesei2, K. Murali2, M. Voetz1, S. Eiden1, A. Duschl3 and E. Madarasz2
1. Bayer Technology Services GmbH, Leverkusen, Germany 2. Hungarian Academy of Sciences, Budapest, Hungary 3. Paris-Lodron-Universität, Salzburg, Austria [email protected]
Engineered amorphous silica nanoparticles (SiO2 NPs) are increasingly used in diagnostics and biomedical research due to their simple production and relatively low costs. SiO2 NPs are considered safe and produced on industrial scale as components of a growing number of commercial products. Despite the potential benefits, there is a concern that exposure to certain types of SiO2 NPs may lead to significant adverse health effects.
The motivation of this study was to investigate the response of different neural tissue-type cells including neural stem cells, neurons and astrocytes to increasing doses of 50 nm fluorescent core/shell SiO2 NPs with different surface functionalization (-NH2, -SH and -PVP). The NPs were deeply characterized using variety of physicochemical methods including zeta potential, DLS, TEM, SEM, AC, BET, XPS, SIMS and XRD due to evaluate the reason of potential toxicity. Results of both the lactate dehydrogenase (LDH) toxicity assays and the
MTT-reduction-based cell metabolism tests indicated that exposure to SiO2 NPs caused cytotoxic damage only at high particle doses. The toxic dose of SiO2 NPs was depended on the chemical composition of the particle surface and also on the type of exposed cells. Both assays revealed that PVP-coated SiO2 NPs did not harm any of the examined cells, even at very high (up to 2 mg/ml) doses. Microscopic studies showed that neurons did not take up particles, while neural stem cells and astrocytes sporadically internalized them. In one-hour exposure, many NPs settled on cell surfaces, but only very few particles entered the cells. The rate of particle accumulation on cell surfaces was reduced in case of the PVP-coated particles.
The data indicated that the PVP-coat on the surface of NPs reduced their toxicity, apparently by lowering the probability of interactions with biomolecules/cellular materials.
This study was supported by the EU 7th framework programme, Marie Curie Actions, Network for Initial Training NanoTOES (PITN-GA-2010-264506).
NanoTOES 1st International Conference 10
Is toxic potential of nanosilver dependent on its size?
Possible mechanisms of toxicity.
Anna Huk1*, Zuzana Magdalenova1, Matthew Boyles2, Paul Schlinker2, Albert Duschl2
Maria Dusinska1
1. Health Effects Laboratory CEE, NILU, Instituttveien 18, 2007 Kjeller, Norway 2. Paris-London University, Hellbrunnerstraße 34, 5020 Salzburg, Austria [email protected]
Because of its antibacterial properties nanosilver is one of the most commonly used engineered nanomaterial (ENMs) [1]. Even of its presence on commercial market since 60 years, its toxic potential is still not well understood. The first toxicology studies have reported controversial results, showing both the toxic, as well as no effects [2].These discrepancies can be partially explained by huge differences in properties of material used and low quality of ENMs commercially available on market. Not accurately done characterization of ENMs is one of main reasons of discrepancy in nanotoxicology study.
The aim of our study was to compare toxic potential of nanosilver with different characteristics (size, shape and surface) which can explain inaccuracies in toxicology results. Detailed characterization of ENMs was performed to measure properties which may influence uptake of ENMs by cells, transport accross biological barriers and toxicity [3].
In our study we were interested, if the silver ENMs synthesized with the same method, the same chemical composition and coating but with different size (50nm, 80nm and 200 nm), can induce various biological responses in mammalian cells cultured in vitro. We measured cytotoxic effects (trypan blue exclusion and relative growth activity assays), DNA strand breaks (the alkaline Comet assay), oxidative DNA damage (enzyme-linked immunosorbent assay and qRT-PCR of antioxidants expression), and mutagenicity (the mammalian in vitro HPRT gene mutation test). Our results show that toxicity of nanosilver can be size dependent and suggest that the cell damage ENMs induced is triggered by different, simultaneous mechanisms.
References
1. M. Ahamed, M. S. AlSalhi, M.K.J. Siddiqui , Clinica Chimica Acta, 2010, 411, 1841–1848 2. S. Wijnhoven, W. Peijnenburg, C.A. Herberts, W.I Hagnes, A.G. OOmen E H.W. Heugens, B. Roszek3, J. Bisschops, I. Gosens, Nanotoxicology, 2009, 2, 109-138 3. B. Fubini, M. Ghiazza, I. Fenoglio, Nanotoxicology, 2010, 4, 347–363
Supported by ITN Marie Curie FP7 NanoToes, ’Nanotechnology: Training of Experts in Safety’, grant agreement no.: 264506.
NanoTOES 1st International Conference 11
Interactions of Ag nanoparticles with organic matter in terrestrial and aquatic microcosms: the role of coatings.
Jason M. Unrine1, Annie Whitley1, Benjamin P. Coleman2, Audrey J. Bone2, Andreas P. Gondikas2, Paul M. Bertsch1 and Cole W. Matson3.
1University of Kentucky, Lexington, KY, USA, 2Duke University, Durham, NC, USA 3Baylor University, Waco, TX, USA
Silver nanoparticles (AgNPs) have been incorporated into a variety of consumer and medical products to confer antimicrobial properties. Silver NPs can be released from consumer products and enter wastewater streams where they will ultimately end up in surface water or in sewage sludge. Sewage sludge is often applied to agricultural land or disposed of in landfills. It is widely acknowledged that water (including pore water) chemistry, pH, ionic strength and organic matter content can affect the transformation, bioavailability and toxicity of Ag NPs. Despite this, few studies have adequately characterized the behavior of engineered nanomaterials in complex environmental testing media. Here we present the results of two investigations of how Ag NPs with different coatings affect environmental matrices with a focus on organic matter.
In the first experiment, either polyvinylpyrrolidone (PVP) coated Ag NPs or gum arabic (GA) coated Ag NPs were incubated in aquatic microcosms containing either water (W), water and sediment (WS), water, sediment and plants (WSP) or water and plants (WP). In the second experiments either PVP-AgNPs or citrate (Cit) coated AgNPs were incubated in microcosms containing a natural sandy loam and either control, 5 or 15% domestic sewage sludge and were incubated for periods of 1 week to 6 months. The samples were analyzed using asymetrical flow field flow fractionation with ultraviolet-visible, laser light scattering and inductively coupled plasma mass spectrometry. Solids from the experiments were analyzed using X-ray absorption spectroscopy.
We found that there were significant interactions between particle surface coating and matrices in aquatic or terrestrial media. In the first experiment, dissolved organic matter (DOM) from plants stabilized PVP-AgNPs as primary particles, while it stimulated the dissolution of GA-AgNPs. In the second experiment we found that Cit-AgNPs were stable as primary particles in soil pore water with no sewage sludge ammendment, but PVP-AgNPs tended to aggregate. Upon addition of sewage sludge, both particle types were extensively transformed and Ag ions were found bound to a fraction that was characteristic of alluminosilicate clay particles. These results highlight complex interactions between particle coating and organic matter in environmental matrices.
NanoTOES 1st International Conference 12
Role of protein corona in uptake, localization and (delayed) impacts of nanomaterials
K. A. Dawson1, F.Wang, A. Salvati 1. Centre for BioNano Interactions, School of Chemistry & Chemical Biology, University College Dublin, Belfield, Dublin 4, Ireland
It is believed that the corona of biomolecules that surround nanoparticles immediately upion contact with biofluids is sufficiently strong that it is the nanoparticle-corona complex that interacts with cells1. We have recently shown that the corona is sufficiently strongly bound to nanoparticles that it is retained during nanoparticle uptake by cells, and that the presence of such a biomolecule corona affects (delays) the impact of the nanomaterials on the cells following uptake and localisation in lysosomes. To this end, we have fluorescently labelled the full bovine serum used for tissue culture by reaction with an Alexa 488 dye on the amino groups of the serum proteins. In this way, a labelled corona was formed and its evolution during nanoparticle uptake and intracellular trafficking by 1321N1 astrocytes was tracked. Using fluorescently labelled amine-modified polystyrene nanoparticles, we could show that the corona proteins are retained, at least in part, during uptake of the nanoparticles, thus generating the first evidence that the corona can enter cells with nanoparticles. Following uptake, time resolved fluorescence imaging indicated that the corona-nanoparticle complexes are also trafficked together to the final nanoparticle destination in the lysosomes. Once there, a progressive decrease of the corona fluorescence could be monitored, probably due to enzymatic degradation of the labelled corona proteins in the lysosomes2. This progressive corona degradation was clearly accompanied by lysosomal swelling and consequent apoptosis, as previously reported for these nanoparticles on these cells (see also Bexiga et al, 20113).
References
1. D. Walczyk, F. Baldelli Bombelli, M. Monopoli, I. Lynch, K.A. Dawson, What the cell “sees” in bionanoscience. J. Am. Chem. Soc., 2010, 132, 5761-5768. 2. V. See, P. Free, Y., Cesbron, P. Nativo, U. Shaheen, D.J. Rigden, D.G. Spiller, D.G. Fernig, M.R.H. White, I.A. Prior, et al., Cathepsin l digestion of nanobioconjugates upon endocytosis. ACS Nano 2009, 3, 2461-2468. 3. M.G. Bexiga, J.A. Varela, F. Wang, F. Fenaroli, A. Salvati, I. Lynch, J.C. Simpson, K.A. Dawson, Cationic nanoparticles induce caspase 3-, 7- and 9-mediated cytotoxicity in a human astrocytoma cell line. Nanotoxicology 2011, 5, 557-567.
NanoTOES 1st International Conference 13
Standardised assay platforms for cell-based nanosafety testing
M. L. Yam, T. A. Duff, J. J. Oliver and C. J. Wilde AvantiCell Science Ltd., GibbsYard Building, Auchincruive, Ayr KA6 5HW, Scotland, UK. [email protected]
The emergence of nanotechnology brings with it a requirement to evaluate the impact of engineered nanomaterials on human health and the environment. To date, there are few standard methods for nanosafety testing or determining therapeutic benefit of nano- structured materials, and many existing methods are either incompletely validated or physiologically-irrelevant, or both. Hence, standardised and validated assay platforms for the testing of nanomaterials are much needed. Cell-based analysis provides a relatively convenient means of testing the effect of engineered nanomaterials on human health. This cell-based testing should have real predictive value; therefore it should be based on the right cells, cultured under the right culture conditions, and using assay readouts that together produce a truly-predictive evaluation of the test material. In practice, this means using human cells in culture microenvironments where tissue-specific functionality is retained, and measured with medium-throughput capacity. In this presentation, we describe the cell culture technology and analytical principles which offer prospect of robust, reproducible and highly- predictive nanotoxicological evaluation.
NanoTOES 1st International Conference 14
Media effects on silver nanoparticle bioavailability and toxicity in Caenorhabditis elegans
Carolin L. Schultz1,2, William I. Tyne1, Stephen Lofts3, Alexandre Vakourov4, Kerstin Jurkschat2, Rudo Verweij5, David J. Spurgeon1, Alison Crossley2, Claus Svendsen1
1. Centre for Ecology and Hydrology, Maclean Building, Benson Lane, Crowmarsh Gifford, Wallingford, Oxon, OX10 8BB, UK 2. Materials Department, University of Oxford, Parks Road, Oxford OX13PH, UK 3. Centre for Ecology and Hydrology, Lancaster Environment Centre, Lancaster LA1 4AP, UK 4. Centre for Molecular Nanoscience (CMNS), School of Chemistry, University of Leeds, LS2 9JT, UK 5. Department of Animal Ecology, Institute of Ecological Science, Free University Amsterdam, de Boelelaan 1085, 1081 HV Amsterdam, Netherlands
Media properties can affect metal fate and behaviour considerably and have successfully been used to predict metal toxicity as well as their bioavailability. To test whether media chemistry can also be used to improve the understanding of silver nanoparticle toxicity, the effect of a range of different synthetic soil pore water characteristics was assessed in Caenorhabditis elegans. Experiments on the influence of Ca2+, Na+, pH or fulvic acid concentration were conducted while the concentrations of the remaining components were kept constant. Exposures measured reproductive toxicity of either silver nitrate @ 0.27 mg/l or silver nanoparticles @ 6.5 mg/l (the EC30 and EC50 from standard medium respectively). Further experiments will be conducted to investigate the interaction of the silver nanoparticles with a biomembrane model including effects on membrane integrity and permeability. The model used is a phospholipid monolayer on a mercury film electrode allowing for interactions of the monolayer with the potentially biomembrane active nanoparticles to be recorded electrochemically.
Of the tested media components only changes in pH had a significant effect on silver toxicity. + Increased H concentration decreased AgNO3 toxicity as predicted, but in contrast increased nanoparticle toxicity. The observation for silver nanoparticles can be attributed to a direct nanoparticle effect. These hypotheses were tested characterizing key material features like nanoparticle size and zeta potential. Together these data indicate that both ionic silver and silver nanoparticle toxicity can be modelled based on input parameters relating Ag concentration and media pH.
NanoTOES 1st International Conference 15
Toxicity and molecular level effects of Au and Ag nanoparticles on soil invertebrates, Caenorhabditis elegans and Eisenia fetida.
O. V. Tsyusko1, J. M. Unrine1, S. Hardas1, W.A. Shoults-Wilson2, B. Collin1, D. Spurgeon3, E. Blalock1, C. Starnes1, D. Starnes1, M. Tseng4, G. Joice1, D. A. Buttrfield1, and P. Bertsch1,
1. University of Kentucky, Lexington, KY, 40546, USA 2. Roosevelt University, Chicago, IL, 60605, USA 3. Centre for Ecology & Hydrology, Wallingford, Oxforshire, OX10 8BB, United Kingdom 4. School of Medicine, University of Louisville, Louisville, Kentucky, 40202, USA [email protected]
Au and Ag nanoparticles (Au-NPs and Ag-NPs) were used for studying molecular level effects of manufactured nanomaterials in soil invertebrates, Caenorhabditis elegans and Eisenia fetida. We used E. fetida to investigate short-term changes in levels of expression of nine stress response genes and oxidative damage of proteins to polyvinylpyrrolidone (PVP) coated Ag-NPs and AgNO3 in natural soils. The responses varied significantly among days with the third day showing the highest number of significant changes in gene expression and protein carbonyls (PC) relative to controls. Similarity in gene expression patterns for Ag-NPs and AgNO3 suggest that ions were primarily responsible for the observed responses. Significant correlations of decreased gene expression with increased Ag soil concentration for two genes (CAT and HSP70) on day three suggest that their genetic responses are determined by Ag soil concentration independent of the NP size or Ag form. Significant increases in levels of PC on day three in all treatments indicate that E. fetida experienced oxidative stress three days after exposure to both NP and ions. Thus, Ag ions seem to be primarily responsible for Ag-NP toxicity in E. fetida. However, given that <15% of Ag in the NPs was oxidized in these soils, dissolution of Ag-NPs is likely to occur after or during their uptake. In another study, Au-NPs were used as a probe for examining particle specific effects in a model organism C. elegans. 797 differentially expressed genes were identified from microarray analyses, and 38 of these genes were associated with seven common biological pathways. Up-regulations of 26 pqn/abu genes from noncanonical Unfolded Protein Response (UPR) pathway and significant increases in sensitivity to Au-NPs in UPR C. elegans’s mutant (pqn-5) and knockout (abu-11) suggest possible involvement of these genes in a protective mechanism against Au-NPs. Differences in distribution of Au in endocytosis mutants (chc-1 and rme-2) versus wild type examined using synchrotron-based X-ray microfluorescence spectroscopy were correlated with differences in toxicity. Meta- analysis comparing our study to four others examining C. elegans’ transcriptional responses to Ag-NP, pathogen, and oxidative stressors revealed a unique Au-NP-specific expression signature.
NanoTOES 1st International Conference 16
Biocompatible micro-sized cell culture chamber for the detection of nanoparticle-induced interleukin-8 promoter activity on a small cell population
Y. Kohl1, A. Duschl2 and H. von Briesen1 1. Fraunhofer Institute for Biomedical Engineering, Ensheimer Str. 48, 66386 St. Ingbert, Germany 2. University of Salzburg, Hellbrunner Str. 34, 5020 Salzburg, Austria [email protected]
Not only within the framework of the European Union Regulation of chemical substances, REACH, there is a growing interest in alternative non-invasive in vitro methods for the risk assessment of nanoparticles (NP)1. Standardized methods for determining toxic effects of NP on cells are commonly based on examining and averaging the response of a cellular population, such as 2D- or 3D cell cultures2. Such averaging might result in mis- interpretations when only individual cells within a population respond to a certain stimulus, due to whether or not they came in contact with NP. Thus, there is a need for new techniques bridging the gap between population analysis and quantitative single cell analysis.
With regard to the development of new non-invasive in vitro methods for REACH or the pharmaceutical screening, this study aimed for the evaluation and validation of a biocompatible micro-sized cell culture chamber for the detection of NP-induced interleukin-8 (IL-8) promoter activity on a small cell population. Biological microelectromechanical systems (Bio-MEMS) present a suitable approach for analyzing a small amount of cells in a defined cell culture area3. In the present study, a micro-sized cell culture chamber (MCC) with a 800 nm thick silicon nitride membrane (cell culture area 0.27 mm2) was developed. The successful proliferation and differentiation of different cell lines (human stem cells, neuronal cells, human lung cells) in the MCC verified its biocompatibility. In combination with a transfected reporter gene cell line the MCC was validated as new non-invasive in vitro system to investigate dose- and time-dependent NP-effects on inflammatory response.
The new MCC-based method can give dynamic information at the level of adherent single cells of a small cell population and presents a new non-invasive in vitro test method for nanotoxicological real time analysis of individual adherent cells in a defined population.
References 1. Regulation EC No. 1907/2006 of the European Parliament and of the Council, 2006. 2. N. Lewinski, V. Colvin and R. Drezek, Small, 2008, 4, 26-49. 3. S. Gotz and U. Karst, Anal. Bioanal. Chem., 2007, 387, 183-192.
NanoTOES 1st International Conference 17
In vitro evaluation of silver nanoparticles of different sizes in assays for inflammation, genotoxicity and developmental toxicity
Margriet Park1,2, H. van Loveren1,2, A.M. Neigh3 L.J.J. de la Fonteyne1, J.P. Vermeulen1, E.R. Gremmer1 and W.H. de Jong1
1. RIVM, Bilthoven, The Netherlands 2. Maastricht University, Maastricht, The Netherlands 3. NanoComposix, San Diego, USA
Silver nanoparticles are currently listed as the most commonly used nanomaterials in consumer products, and their antibacterial properties are of interest for medical applications. In light of the anticipated increased human exposure to silver nanoparticles, evaluation of their potential adverse effects is necessary. We have studied well-characterized silver nanoparticles of 20, 80 and 113 nm in vitro, using assays for cytotoxicity, inflammation, genotoxicity and developmental toxicity.
Figure 1: Metabolic activity of L929 fibroblasts (A) and RAW 264.7 macrophages (B) as a function of concentration of silver nanoparticles.
Silver nanoparticles induced effects in all endpoints studied, but effects on cellular metabolic activity (Fig. 1) and membrane damage were most pronounced. In all toxicity endpoints studied, silver nanoparticles of 20 nm were more toxic than the larger nanoparticles. Collectively, our results indicate that the ability of silver nanoparticles to inflict cell damage may result in effects on various other toxic endpoints. In addition, the potency of silver in the form of nanoparticles to induce cell damage compared to silver ions is cell type and size- dependent.
NanoTOES 1st International Conference 18
Nano-immunosafety: issues in assay validation
Diana Boraschi1, Cristina Battaglia2, Silvio Bicciato3, Luca Costantino3, Linda Stöhr4, Yang Li1, Albert Duschl5, Eudald Casals6, Victor F. Puntes6, and Paola Italiani1 1. National Research Council, Pisa, Italy 2. University of Milano, Milano, Italy 3. University of Modena and Reggio Emilia, Modena, Italy 4. Grimm AerosolParis-Lodron Universitaet Salzburg, Salzburg, Austria [email protected]
Immunosafety of nanomaterials is a major health issue that should be addressed with appropriate and reliable tools. Possible interference of nanoparticles with the normal functions of the immune system, in particular innate/inflammatory responses, may lead to pathological consequences. Nano-particulate matter can trigger reactions similar to those initiated by infectious microorganisms. The extent and duration of the inflammatory reaction dictates the biological outcome, i.e. physiological resolution or persisting inflammation eventually leading to pathology.
To assess the immunosafety of nanomaterials, robust and representative assays that can predict the immune-related risk of developing diseases should be designed and implemented. These should be: rapid and easy to perform (e.g., in vitro assays); take into account the physico-chemical characteristics, biological contamination and behaviour of nanoparticles (e.g., optical interference with assay readouts); relevant and predictive of the nanoparticle effects in vivo in human beings (e.g., by using human primary cells in vitro); reliable (as defined by the Klimisch score), robust and reproducible (e.g., after standardisation with cell lines and selected reported genes).
An in vitro assay based on human primary blood monocytes has been set up and validated, which recapitulates the different phases of the defence response, from recruitment to development of inflammation, resolution, repair and re-establishment of homeostasis. Genome-wide transcriptomic analysis and multiplex proteomic detection of inflammatory factors have been used for profiling the response in the absence of interfering molecules (“physiological” defence response) and defining “signatures” that describe its evolution. These signatures will then be used for monitoring, in comparative experiments, the possible alterations caused by nanoparticles. Simplification of the assay involves identification of selected biomarkers (e.g. the cytokine pair IL-1 /IL-1Ra) and the possible set up of suitable reporter cell lines. Preliminary experiments with metal nanoparticles show little/no interference with the development of the physiological defence response.
In conclusion, the design and implementation of an array of relevant immunosafety tests is to be considered as one of the basic steps necessary to the sustainable knowledge-based development of nanotechnologies applied to medicine.
NanoTOES 1st International Conference 19
Poster Abstracts
NanoTOES 1st International Conference 20
Synthesis, Surface Modification and Immunological properties of Peptide- conjugated Gold Nanoparticles
S. Rubio1, N. G. Bastús1 and V. F. Puntes1, 2, 3
1. Institut Català de Nanotecnologia (ICN), Campus UAB, 08193 Bellaterra, Barcelona, Spain. 2. Universitat Autònoma de Barcelona (AUB), Campus UAB, 08193 Bellaterra, Barcelona, Spain. 3. Institut Català de Recerca i Estudis Avançats (ICREA), 08010 Barcelona, Spain. [email protected]
Although the remarkable rapid development of synthetic and functionalization protocols for the use of inorganic nanoparticles (NPs) in biomedical applications, relatively little is known about NP's behaviour in complex biological systems. Thereby it is particularly true for the immune system, which is responsible for maintaining body integrity, detecting and categorizing self and non-self molecules in order to protect the host from succumbing to infections.
Herein we explore the use of engineered inorganic nanoparticles as substrates to carry multifunctional ligands to manipulate the immune system in a controlled manner via its interaction with specific cell-membrane receptors1-3. For this purpose, we synthesized highly monodispersed citrate-stabilized gold NPs following a kinetically-controlled seeded growth strategy4. As-synthesized NPs were further functionalized with peptides by following a ligand exchange via the thiol group of the cysteine moiety. Obtained samples were characterized by UV-Vis spectroscopy, Dynamic Light Scattering and Z-Potential in order to determine the success of the conjugation process as well as the stability and aggregation state of final colloidal solutions. Special efforts were devoted to study the conformation structure of the peptide layer onto the gold NP's surface by using monitoring the etch resistance to sodium cyanide digestion5. Obtained results allowed us not only to determine the robustness of functionalized NPs but also to study the degree of order of the peptide shell, a key factor when correlating final structure of conjugated NP with biological effects.
References
1. N. G. Bastús, E. Sánchez-Tilló, S. Pujals, C. Farrera, C. López, E. Giralt, A. Celada, J. Lloberas and V. Puntes, ACSNano, 2009, 3(6), 1335-1344. 2. I. Hamad, O. Al-Hanbali, A. C. Hunter, K. J. Rutt, T. L. Andresen and S. M. Moghimi, ACSNano 2010, 4(11), 6629-6638. 3. S. M. Moghimi, D. Peer and R. Langer, ACSNano, 2011, 5(11), 8454-8458. 4. N. G. Bastús, J. Comenge, V. F. Puntes, Langmuir, 2011, 27, 11098-11105. 5. B. C. Mei, E. Oh, K. Susumu, D. Farrell, T. J. Mountziaris, and H. Mattoussi, Langmuir 2009, 25(18), 10604–10611.
NanoTOES 1st International Conference 21
Experimental Protocol Design for Reactivity Test for Nanoparticles and Characterization of the Reactive Oxygen Species from Brake Ware Particles
J. Zhao1 and M. Riediker1 1. Institut de Santé au Travail, Rue du Bugnon 21, CH-1011 Lausanne, Switzerland
Inhalation of Nanoparticles (NPs) could lead to serious health problems. Understanding the reactivity of NPs could substantially contribute to clarifying the toxicity of these particles with special natures. However, the lack of a uniform and effective approach has led only a few studies have been finished. In this study, we have been able to present a reliable method to detect reactivity of NPs. We evaluated several commonly used chemicals to prepare the working solution with typical concentrations. We also compared different sonication methods to disperse NPs. Moreover, ultra filtration has been investigating to overcome the difficulties on using high sample concentrations. Scan absorbance spectrum over time was made to support our conclusion on filtration. A 2’7-dichlorodihydrofluorescin (DCFH) working solution with 0.5 unit/ml horseradish peroxidase (HRP) yields repeatable results. Also, ethanol was chosen to prepare DCFH stock solution. Sodium buffer was preferred as the solvent for DCFH-HRP working solution. Based on our results, sonication in DCFH-HRP working solution would provide better data than using a buffer or Tween 80 as the sonication solution. Moreover, particle concentration in the magnitude of mg/ml was considered too high. We recommend ultra filtration to avoid signal block of the particle samples. The protocol could be used as a standard method in both academic and industry research and yields reliable and repeatable results. We are currently applying this approach to detect reactivity of several kinds of break wear particles.
NanoTOES 1st International Conference 22
Oxidative stress in response to nanoparticle surface charge
Paul Schlinkert1, Matthew Boyles1, Eudald Casals2, Victor Puntes2, Albert Duschl1 1. University of Salzburg, Salzburg, Austria 2. Institut Catala de Nanotecnologia, Barcelona, Spain [email protected]
Over the last years the application of nanomaterials has become more widespread. Gold nanoparticles (NP) are increasingly found in the medical sector, e.g. in drug delivery systems1, but also in cosmetics, food packaging and beverages2. Silver NP are now present in socks, storage bags, chopping boards, surface disinfectants and throat sprays3,4. Due to this, both consumers and workers involved in the production of these particles are at a higher risk of NP exposure. Analysis of potential NP toxicity and their ability to induce oxidative stress (OS) via the production of reactive oxygen species (ROS) is therefore needed.
For this study, Au and Ag NP have been coated with different surfactants to achieve different surface charges, ranging from a single negative charge to three increasingly positive charged particles with a size range of 7-10 nm. Their cytotoxicity and oxidative potential were studied in A549 cells.Cell viability was assessed via the cell titer blue (CTB) assay, and cytotoxicity via the lactate dehydrogenase assays (LDH). The expression of key markers of oxidative stress such as superoxide dismutase 1 (SOD1) and glutathione peroxidase 1 (GPx1) were analysed by RT-PCR. ROS production was measured with the DCFH-DA assay.
During this study minor cytotoxicity was observed after exposure to negatively or low positively charged particles, irrespective of the particle composition. Cytotoxicity did however increase when cells were exposed to particles with a high positive charge. ROS production in response to particle exposures followed the same trend. These findings are in line with those observed by Bhattacharjee et. al5, who studied the effects of oppositely charged silicon NP in macrophages.
References
1. E. Girgis, W.K.B. Khalik, A.N. Emam, M.N. Mohamed, K.V.Rao, Chem. Res. Toxicol, 2012, 25(25), 1086-1098. 2. J.H. Sung, J.H. Ji, J.D. Park, M.Y. Song, K.S. Song, H.R. Ryu, J.U. Yoon, K.S. Jeon, J. Jeong, B.S. Han, Y.H. Chung, H.K. Chang, J.H. Lee, D.W. Kim, B.J. Kelman, I.J. Yu, Part. Fibre.Toxicol, 2011, 8(16) 3. K. Chaloupka, Y. Malam, A.M. Seifalian, Trends. Biotechnol, 2010 28 (11) 4. M.E. Quadros, L.C. Marr, Environ. Sci. Technol, 2011 45 /24), 10713-9 5. S. Bhattacharjee, L.H. de Haan, N.M. Evers, X. Jiang, A.T. Marcelis, H. Zuilhof, I.M. Rietjens, G.M. Alink, Part. Fibre. Toxicol, 2010, 7 (25)
This study was supported by the EU 7th framework programme, Marie Curie Actions, Network for Initial Training NanoTOES (PITN-GA-2010-264506)
NanoTOES 1st International Conference 23
Study of silver nanoparticle-induced production of IL-8 and ROS in human lung cells
S. A. Q. Shah1, Y. Kohl1, T. Knoll1, Albert Duschl2 and H. von Briesen1 1. Fraunhofer Institute for Biomedical Engineering IBMT, Ensheimer Str. 48, 66386 St. Ingbert, Germany 2. University of Salzburg, Hellbrunner Str. 34, 5020 Salzburg, Austria [email protected]
Due to their antimicrobial activity silver nanoparticles (AgNPs) are used in a widespread range of industrial, medical and consumer products. The physiochemical properties of the nanoparticles (NPs) can potentially cause adverse effects on tissues or organs1, 2. At the moment there is a serious lack of information concerning the biological activity of nanosized silver in human tissue cells. In this study, we analyzed the interleukine-8 (IL-8) production and the production of reactive oxygen species (ROS) in human lung epithelial carcinoma cells (A549) after their exposure to AgNPs. The cells were incubated with 10 nm and 20 nm sized AgNPs (1 x 1011 NP/ml to 6.25 x 109 NP/ml) for 4 h and 24 h. Afterwards the IL-8 production, a marker for inflammatory response, was determined via enzyme- linked immunosorbent assay (ELISA). Further the induction of ROS was quantified. Beside the analysis of the nanoparticle-induced inflammatory response via ELISA-assay the human IL-8 induction was investigated using the transfected reporter gene cells pIL-8 GFP A549 and pIL-8 Luc A549.The pIL-8 GFP A549 cells are transfected with the green fluorescence protein (GFP) reporter gene and the pIL-8 Luc A549 cells with Luciferase (Luc) reporter gene. The GFP-expression, as well as the luciferase activity correlates with the inflammatory status of the cells. All tested cell lines did not show any evidence of AgNPs induced IL-8 production after 4 h and 24 h exposure. Using the pIL-8 Luc A549 cells no AgNP-induced IL- 8 gene expression was determined up to the highest tested NP concentration. The AgNP- treatment of pIL-8 GFP A549 cells also did not induce an increase in GFP expression and thus no increase in inflammatory cell response. Quantifying the IL-8 concentration in the supernatant of the A549 cells also resulted in no concentration-dependent increase in the IL- 8 production after AgNPs exposure. Determining the ROS production by flow cytometry resulted in an oxidative potential of the AgNPs. The results of this study verify no inflammatory effect of the tested 10 nm and 20 nm sized AgNPs in human lung epithelial cells upto 6.25 x 109 NP/ml after 24 h exposure.
References 1. A. M. Schrand, M. F. Rahman, S. M. Hussain, J. J. Schlager, D. A. Smith and A. F. Syed, WIREs. Nanomed. Nanobiotechnol., 2010, 2, 544–568 2. J. Aaseth, A. Olsen, J. Halse and T. Hovig, Scand J. Clin Lab Invest., 1981, 41:247-251
NanoTOES 1st International Conference 24
The on-site assessment of occupational nanoparticle exposure: general problems and possible solutions
Linda C. Stoehr1, 2, Albert Duschl2, Markus Pesch1 1. GRIMM Aerosol Technik GmbH & Co. KG, Dorfstraße 9, 83404 Ainring, Germany 2. University of Salzburg, Hellbrunner Str. 34, 5020 Salzburg, Austria [email protected]
One of the main routes for nanoparticles (NPs) to enter the body is via inhalation. Workers and consumers may inhale high doses of NPs during an accident scenario or through exposure for prolonged periods of time, making airborne nanoparticles an important group of NPs to consider when testing for NP-induced health effects. There are many tests for assessing the biological impact of nanoparticles1. Most are laboratory-based, need cell culture facilities and specially trained personnel and therefore cannot be conducted on site and require the collection and transport of the material. However, the physicochemical characteristics of the NPs may change during the collection and transport process2, which in turn makes it difficult to attribute the effects seen in the laboratory to the conditions on site. Furthermore, efficient deposition of particles from the air onto cells in conventional culture is difficult to achieve due to the high surface tension of the media. In order to circumvent these problems, we are currently developing biological assays to assess potential health hazards of airborne NPs for on-site application. The proposed device will make use of stably transfected reporter cell lines3 directly exposed to the airborne nanoparticles, with cytotoxicity and immune responses detected in real-time. To closely mimic the conditions in the lung, the cells will be exposed to the airborne NPs either at the air-liquid interface4,5 or under slightly submerged conditions with additives that facilitate the transition of particles from the air to the liquid phase by reducing the medium’s surface tension6. This approach is expected to give a more representative evaluation of potential health hazards of NP exposure than the existing lab-based methods.
References
1. Kroll, M.H. Pillukat, D. Hahn and J. Schnekenburger, Eur. J. Pharm. Biopharm., 2009, 72, 370- 377. 2. A.L. Holder, D. Lucas, R. Goth.Goldstein and C.P. Koshland, Toxicol. Sci. 2008, 103, 108-115. 3. G.J. Oostingh, M. Schmittner, A.K. Ehart, U. Tischler and A. Duschl, Toxicol. In Vitro, 2008, 22, 1301-1310. 4. H.R. Paur, S. Mülhopt, C. Weiss and S. Diabaté, Journal of Consumer Protection and Food Safety 2008, 3, 319-329. 5. A.G. Lenz, E. Karg, B. Lentner, V. Dittrich, C. Brandenberger, B. Rothen-Rutishauser, H. Schulz, G.A. Ferron and O. Schmid, Part. Fibre. Toxicol., 2009, 6: 32. 6. S. Schürch, M. Lee and P. Gehr, Pure & Appl. Chem., 1992, 64, 1745-1750
This study is being supported by the EU 7th framework programme, Marie Curie Actions, Network for Initial Training NanoTOES (PITN-GA-2010-264506).
NanoTOES 1st International Conference 25
Inorganic nanoparticles-protein corona: formation and controlling factors.
N. T. T. Tran1, 2, M. Boyles3, P. Schlinkert3, A. Duschl3, E. Casals1 and V. F. Puntes1, 2 1. Institut Català de Nanotecnologia, Barcelona, Spain 2. Universitat Autònoma de Barcelona, Barcelona, Spain 3. University of Salzburg, Salzburg, Austria [email protected]
Nanoparticles (NPs), due to their unique physical and optical properties, have been increasingly used in a great deal of biological applications, particularly in medicine. Therefore, an understanding of interactions at the nano-bio interface plays a crucial role especially in term of nanosafety.1 Actually, once they come into contact with biological fluids, NPs undergo surface modifications which may, in turn, lead to the alteration of circulation parameters, organ uptake and internalization of NPs as well as their potential cytotoxicity. One of the most significant modifications is the formation of protein corona which results from the competitive adsorption of proteins onto NP´s surface.2, 3 Being governed by Vroman effect in general, protein corona is a time-dependent process in which higher mobile proteins approach NP´s surface first and then are replaced by proteins with higher affinity.2, 3 In addition, it has been found that NP-protein interactions vary with not only particle composition but also particle surface including radius of curvature, charge, coatings. As a result, NP-protein corona can be controlled by modifying these parameters.
In our work, we investigate the protein corona formation by exposing NPs which are different in composition and surface properties to a variety of cell culture media. Temporal adsorption of proteins is characterized using UV-Vis, Z-potential and Dynamic Light Scattering (DLS). The formation of different NP-protein corona is confirmed by a red shift in surface plasmon resonance band along with a drop in surface charge and an increase in hydrodynamic diameter.
References
1. A. E. Nel, L. Madler, D. Velegol, T. Xia, E. M. V. Hoek, P. Somasundaran, F. Klaessig, V. Castranova and M. Thompson, Nature Mater., 2009, 8, 543–557.
2. E. Casals, T. Pfaller, A. Duschl, G. J. Oostingh and V. Puntes, ACS Nano, 2010, 7, 3623-3632.
3. E. Casals, T. Pfaller, A. Duschl, G. J. Oostingh and V. Puntes, Small, 2011, 7, 3479-3486.
NanoTOES 1st International Conference 26
Gold Nanoparticles as Drug Delivery Agents for Cisplatin: Avoiding Side Effects of Chemotherapy.
J.Comenge1, C.Sotelo2, F.Romero3, F.Domínguez2, V. Puntes1 1.Catalan Institute of Nanotechnology (ICN) and Universitat Autònoma de Barcelona. 2. Faculty of Medicine, Santiago de Compostela University (USC). 3. Molecular Science Institute, University of Valencia (UV). [email protected]
Cisplatin is the most used chemotherapeutic agent in many types of cancers. Here we show that cisplatin-induced toxicity, which is the main limiting factor for chemotherapy, is clearly reduced without affecting the therapeutic benefits of the drug. This is achieved by attaching a cisplatin derivative to AuNPs via a pH-sensitive coordination bond. This is related to the change on the biodistribution as well as the different processing of the drug when it is attached to gold nanoparticles. Nanoparticles not only act as a delivery agent, but protect the drug from being deactivated by plasma proteins until they are internalized via endocytosis and cisplatin is released. The possibility of tracking the drug and the vehicle separately enables a better understanding on how nanocarriers are processed by the organism.
Figure 1. Biodistribution of the drug is modified after conjugation to AuNPs, avoiding organs where it is known to induce toxicity. Hystological analysis revealed the normal appearance of the kidneys after receiving the AuNPs-cisplatin treatment, while cisplatin-induced acute nephrosis is observed in the case of treatment with the free drug. On the contrary, AuNPs that do not reach the tumour are mainly accumulated in organs of the RES where no toxicity is observed. Further details on the other organs and controls are also reported.
NanoTOES 1st International Conference 27
Impact of nanoparticles’ shape and medium composition on silver nanoparticle toxicity.
Bogumiła Reidy, Kenneth A. Dawson, Iseult Lynch Centre for BioNano Interactions, University College Dublin, Belfield, Dublin 4, Ireland
In the recent years the potential toxic effects of silver nanoparticles (AgNPs) have become a serious concern. As stated in the literature, the stability of AgNPs strongly influences their biological impact - especially their dissolution rate, as silver ions are considered as the main (or one of the main) factor(s) of AgNPs toxicity. Due to the complex chemistry of silver in aqueous solution, the amount of free, toxic silver ions depends, among the other factors, on the composition of the medium. Silver ions can be bound by sulphides and chlorides, forming insoluble complexes, which reduces their bioavailability (toxicity).
Here we present preliminary results on our cytotoxicological studies performed with AgNPs of different shapes (spheres and wires) in two kinds of cell culture media. The results suggest that both nanoparticles shape and media composition had a significant impact on AgNPs toxicity. Nanowires showed higher toxicity than equivalent mass of nanospheres (which could be caused by different dissolution rates), although the toxicity of both was much lower than the toxicity of silver ions (added as AgNO3).
References
N. W. H. Adams and J. R. Kramer Environ Toxicol Chem, 1999, 18, 2667–2673
M. A. Chappell, L. F. Miller, A. J. George, B. A. Pettway, C. L. Price, B. E. Porter, A. J. Bednar, J. M. Seiter, A. J. Kennedy and J. A. Steevens Chemosphere, 2011, 84, 1108–1116. X. Yang A. P. Gondikas, S. M. Marinakos, M. Auffan, J. Liu, H. Hsu-Kim and J. N. Meyer Environ. Sci. Technol., 2012, 46, 1119-1127
NanoTOES 1st International Conference 28
MEETING LOCATION:
Lecture Theatre C006, Health Sciences Centre, University College Dublin
Building Number 29 on the attached map.
NanoTOES 1st International Conference 29
Building Index No. Grid Building Index (cont) No. Grid UCD College of Science No.
UCD School of Biological & Environmental Science 13,18,22,11
tillorgan
Agnes McGuire Social Work Building Richview Buildings Labatory 52 E1 S UCD School of Biomolecular & Biomedical Science 13,18,22,11 Dundrum
(Arts Annexe) 1 E9 hview Lecture Building 53 F1
Ric UCD School of Chemistry & Chemical Biology 13,18,22,11
US AMP C
oatstown UCD Agriculture and Food Science Centre 2 D7 hview Library 54 E1 G
Ric UCD School of Computer Science and Informatics 6,17,65
B K K KROC
Ardmore Annexe 3 C8 hview Memorial Hall 55 F1 LAC
Ric UCD School of Geological Sciences 65,66
errion
Ardmore House 4 C8 Richview Newstead Block A 56 F2 M
ount ount UCD School of Mathematical Sciences 6,34, 64 M
Bank, AIB 5 C8 Richview Newstead Block B (Main Bld) 57 F2 rbour
UCD School of Physics 6, 22, 65 A
Blackrock
indy Belfield Office Park 6 D2 Richview Newstead Block C 58 E3 W
Belgrove Student Residences 7 E8 Richview School of Architecture 59 E1
UCD College of Business & Law No. AMPUS Bicycle Shop 8 B10 ebuck Castle 60 G11 C
Ro UCD School of Business 6, 49
UCD Bowl 9 C4 ebuck Hall Residence 61 F11 BELFIELD
Ro UCD School of Law 60, 73 ooterstown B
Campus Services 10 D7 Roebuck Offices 62 G11 1
1
Milltown UCD Centre for Molecular Innovation D Rosemount Environmental Research N
UC UCD Michael Smurfit Graduate Blackrock
and Drug Discovery 11 D6 Station 63 H4 Hospital
lonskeagh C
Vincents t
Business School Campus S Mast
TE Centre for Research in UCD Science Centre (Hub) 64 D6 R
Infectious Diseases (CRID) 12 B8 UCD Science Centre (North) 65 C6
UCD College of Health Sciences No. Merrion
Centre for Synthesis and UCD Science Centre (West) 66 D6 Donnyrook
ay B ublin UCD School of Medicine and Medical Science 12, 18, 29 D
Chemical Biology (CSCB) 13 D7 UCD Science Centre (East) 67 C6
erbert Park Park erbert
UCD School of Nursing, Midwifery H Charles Institute 14 C5 D Sports Centre 68 E5 Ranelagh UC and Health Systems 29
UCD Clinton Centre for American Studies St. Stephens 69 C10
allsbridge UCD School of Public Health, Physiotherapy and B
(Belfield House) 15 B10 UCD Student Centre 70 D5
E ARLSFORT TERRACE ARLSFORT andymount Population Science 18,29,58,78 S UCD Computer Centre 16 C5 UCD Student Club 71 D9 UCD Computer Science and D Student Learning Leisure and UC UCD College of Agriculture, Food Science and Informatics Centre 17 C6 Recreation Facility 72 E5
Veterinary Medicine No. UCD Conway Institute 18 B5 D Sutherland School of Law 73 D10 Green Stephens St UC UCD School of Agriculture and Food Science 2, 63 Crannóg House 19 G12 rney Building (Administration Building) 74 C8 Tie UCD School of Veterinary Medicine 76, 77 Daedalus Building 20 C9 UCD Urban Institute of Ireland (UII) 75 F1 Energy Centre 21 F3 Veterinary Hospital 76 B6
UCD Engineering & Materials Science Centre 22 C9 UCD Veterinary Sciences Centre 77 B6 Campus Information p a M n o i t a c o Environmental Protection Agency 23 E1 Woodview House 78 B5 L UCD Geary Institute (Arts Annexe) 24 F9 Services Gerard Manley Hopkins Centre (UCD Academic Index International Office) 25 D9 UCD College of Human Sciences No. Bank 5 C8 Glebe House 26 G11 UCD School of Applied Social Sciences 28 Bicycle Shop 8 B10 Glenomena Student Residences 27 C11 UCD School of Economics 41 Campus Bookshop 34 D7 Hanna Sheehy-Skeffington Building UCD School of Education 62 Centra Supermarket 37 D11 Belfield (Arts Annexe) 28 E9 UCD School of Geography Planning Copi-Print 34,41,49 D7,D8,D9 Health Sciences Centre 29 C5 and Environmental Policy 41, 48, 59 Laundry 27, 61 C11, F11 Campus UCD Humanities Institute Ireland 30 F9 UCD School of Information and Library Studies 34 Pharmacy 70 D5 Map Information Point 31 B8 UCD School of Philosophy 41 Post Office 51 D9 UCD Institute of Sport & Health / UCD School of Politics and International Relations 41 Sports Centre Barber 68 E5 Leinster Rugby 32 F2 UCD School of Psychology 41 Student Desk 74 C8 Irish Institute for Chinese Studies UCD School of Social Justice 41 Student Health Service 70 D5 (UCD Confucious Institute) 33 G11 UCD School of Sociology 34, 41 Students' Union 70 D5 UCD James Joyce Library 34 D7 Students' Union Shop 22,34,64 C9,D7,D6 UCD John Hume Institute for Global Irish Studies UCD College of Arts & Celtic Studies No. UCD HR 62 G11 (William Jefferson Clinton Auditorium) 35 B9 UCD School of Archaeology 19, 41, 60 Medical Bureau of Road Safety (MBRS) 36 D5 UCD School of Art History and Cultural Policy 41 Traffic Calming Programme Merville Student Residences 37 D11 UCD School of Classics 41 National Hockey Stadium 38 D4 UCD School of English, Drama and Film 28, 41, 44 Traffic Restrictions in Operation National Institute for Bioprocessing Research UCD School of History and Archives 30, 34, 41 Mon-Fri Barriers closed from: 07.00-10.30 and Training (NIBRT) 39 C12 UCD School of Irish, Celtic Studies, 16.00-19.30 National Virus Reference Laboratory (NVRL) 40 C8 Irish Folklore and Linguistics 41 Newman Building 41 D8 UCD School of Languages & Literatures 41, 44 Gates Opening Times NovaUCD 42 B12 UCD School of Music 41 Oakmount Créche 43 G6 N11 Entrance 24 hours UCD O’Kane Centre for Film Studies UCD College of Engineering and Architecture No. Clonskeagh Entrance, (Mon-Sun) 07.00-00.00 (Observatory) 44 F7 UCD School of Architecture 48,52,54,57,59 Owenstown Entrance (Mon-Sat) 07.00-00.00 O'Reilly Hall 45 C7 UCD School of Biosystems Engineering 2, 22 Fosters Avenue Entrance 07.00-00.00 UCD Unicare: Our Lady Seat of Wisdom Church 46 E6 UCD School of Chemical & Bioprocess Engineering 22 Richview Entrance (Mon-Fri) 07.00-00.00 Pavillion 47 D4 UCD School of Civil, Structural and (Sat) 07.00-18.00 Planning and Environmental Policy 48 E1 Environmental Engineering 48,52,54,57,59 Richview Newstead Gate (Mon-Sun) 24 hours our campus, our care... UCD Quinn School of Business 49 D9 UCD School of Electrical, Electronic and Roebuck Castle, Pedestrian Route 24 hours UCD Research 50 C8 Communications Engineering 6, 22 Greenfield Park, Pedestrian Route 24 hours Emergency Line: Restaurant 51 D9 UCD School of Mechanical & Materials Engineering6, 21 Roebuck Road Gate Pedestrian Route (Mon-Fri) 07.00-18.00 (01 716) 7999 Buildings under construction or in the planning stage are shown in Italics Traffic Barrier Fixed Traffic Barrier Belfield Campus Map, July 2011 Primary Vehicle Route Secondary Vehicle Route Pedestrian Route Woodland walk
< Do nny br Dublin Bus ook Aircoach Bus Stop
N11 Entrance Wexford >
31
76 42 78 8 15 35 18 12 Greenfield Entrance 77 45 3 Vehicle Route 40 27 17 4 22 14 16 5 69 Fosters 65 67 Avenue 74 39 Entrance 20 9 29 50 47 64 13 Belfield Office Park 36 66 6 11 34 41 51 70 2 10 25 71 73 37 38 49 72 46
68 23 e u n 52 e 48 v A
s r e 1 t 28 s o 7 F 54 58 59 75 24 21 53 30 61 Owenstown 55 56 Entrance Richview 57 44 Entrance 32
C lon sk ea 33 gh 62 Ro ad
Clonskeagh 60 Entrance 43 19 Richview Newstead Entrance 26
Rosemount Roebuck Castle Entrance North 63 Roebuck Road
Roebuck Road Gate
PARTICIPANT LIST:
Participant Institution Email Address
ABAD, Silvia European Commission [email protected]
BORASCHI, Diana Instituto di Tecnologie Biomediche [email protected]
COMENGE, Joan The Catalan Institute of Nanotechnology [email protected]
COOKE, Laura University College Dublin [email protected]
COSTANTINI, Maria Health Effects Institute [email protected]
CRONIN, Patrick University of Limerick [email protected]
DAWSON, Kenneth University College Dublin [email protected]
DUSCHL, Albert University of Salzburg [email protected]
DUSINSKA, Maria Norwegian Institute for Air Research [email protected]
EPPACHER, Elisabeth University of Salzburg [email protected]
HAASE, Andrea The Federal Institute for Risk Assessment (BfR) [email protected]
HINDS, David University College Dublin [email protected]
HUK, Anna Norwegian Institute for Air Research [email protected]
IZAK, Emilia Bayer Technology Services GmbH [email protected] KENESEI, Kata Institute of Experimental Medicine [email protected]
KNOLL, Thorsten Fraunhofer Institute for Biomedical Engineering IBMT [email protected]
KOHL, Yvonne Fraunhofer Institute for Biomedical Engineering IBMT [email protected]
KOWAL, Katarzyna University of Limerick [email protected]
KUMARASAMY, Murali Institute of Experimental Medicine [email protected]
LI, Yang Institute for Biomedical Technologies(ITB) [email protected]
LOPEZ, Cecilia The Catalan Institute of Nanotechnology [email protected]
LYNCH, Iseult University College Dublin [email protected]
MADARASZ , Emilia Institute of Experimental Medicine [email protected]
MAGNO, Luis Miguel University College Dublin [email protected]
MOVIA , Dania Trinity College Dublin [email protected]
MULVIHILL, Anne Athlone Institute of Technology [email protected]
O'CONNELL, Ann University College Dublin [email protected] National Institute for Public Health and the
PARK, Margriet Environment [email protected]
PAULUHNM, Jürgen Bayer HealthCare [email protected]
PRINA MELLO, Adriele Trinity College Dublin [email protected]
PUNTES, Victor The Catalan Institute of Nanotechnology [email protected] syed.abdul.qadir.shah@
QADIR SHAH, Syed Abdul Fraunhofer Institute for Biomedical Engineering IBMT ibmt.fraunhofer.de
QUINN, Susan University College Dublin [email protected]
REIDY, Bogumila University College Dublin [email protected]
RIEDIKER, Michael Institute for Work and Health [email protected]
RUBIO, Sofia The Catalan Institute of Nanotechnology [email protected]
SCHLINKERT, Paul University of Salzburg [email protected]
SCHULTZ, Carolin Centre for Ecology and Hydrology [email protected]
SVENSDEN, Claus Centre for Ecology and Hydrology [email protected]
STOHR, Linda Corina GRIMM Aerosol Technik GmbH [email protected] NanoTOES 1st International Conference 30
TRAN, Ngoc The Catalan Institute of Nanotechnology [email protected]
TSYUSKO, Olga University of Kentucky [email protected]
UNRINE, Jason University of Kentucky [email protected]
VOETZ, Matthias Bayer Technology Services GmbH [email protected]
VON BRIESEN, Hagen Fraunhofer Institute for Biomedical Engineering IBMT [email protected]
WAN, Sha University College Dublin [email protected]
WILDE, Colin AvantiCell Science Ltd [email protected]
WOJDYLA, Mateusz University College Dublin [email protected]
YAM, Mun Li AvantiCell Science Ltd [email protected]
ZHAO, Jiayuan Institute for Work and Health [email protected]
NanoTOES 1st International Conference 31