Table of contents

This volume contains a collection of contributions presented at the 4th International Conference on Safe Production and Use of Nanomaterials (NANOSAFE 2014) held in Grenoble, France, from 18th to 20th November 2014.

The issues of fast progress in the field of Nanosafety are up to the potential benefits that nanotechnology can bring to mankind. Making more efficient - more sustainable - easier to share mineral resources, increasing the yields of new energy technologies, enabling drugs that act selectively and locally are just few examples of the wide range of nanomaterial applications that currently benefit humanity. Nevertheless, the dynamic development of nanomaterials requires the adhesion from the general public who rightly demand major progresses in Nanosafety as a prerequisite.

This is our exciting responsibility and challenge!

Following the successful outcome of the three past international conferences on safe production and use of nanomaterials: Nanosafe 2008, 2010 and 2012, the organizing committee has the pleasure to welcoming you again to Minatec, Grenoble with some of the most famous specialists in the field. This year, two new topics have been added dealing with the "New Application of Nanomaterials" and "Nano-responsible Development" in addition to the usual issues addressed in previous Nanosafe conferences such as Expology, Detection and Characterization, Toxicology, Environmental Interactions, Nanomaterials Release, Life Cycle Analysis, Regulation and Standardization, Risk Management.

The debates in 2012 proved highly successful so this formula has been kept in 2014 with 3 round tables: Nano-Responsible Development, Risks and Benefits for the Environment, Toxicology Progress.

In this 4th edition, there were more than 330 registered participants from 28 different countries including 160 oral presentation covering the whole Nanosafety issues in 12 sessions, satellite workshops and round tables. This high number of participants makes this edition one of the most successful conference of the series.

Finally, 2014 was for the organizing committee, a great year as we ramp up the Nano Safety "PNS" platform at Minatec, with a brand new building spread out over 2000 m² of laboratories and more than 50 doctors and engineers dedicated to Nanosafety.

F. Tardif, J-F. Damlencourt, F. Schuster and V. Gaultier

All papers published in this volume of Journal of Physics: Conference Series have been peer reviewed through processes administered by the proceedings Editors. Reviews were conducted by expert referees to the professional and scientific standards expected of a proceedings journal published by IOP Publishing.

Because of high occurrence of side effects of conventional anticancer drugs, new therapies have been widely studied, such as the use of non-usual molecules and the association with nanomaterials. In this context, sildenafil is a molecule that has been lately considered as a co-adjuvant in cancer therapy because it inhibits an enzyme with a potential role in tumor progression, phosphodiesterase-5. Taking into account the combination of nanoparticles and sildenafil, we produced systems based on mesoporoussilica nanoparticles, Pluronic F-127 and sildenafil as the main therapy for prostate cancer.

Superparamagnetic iron oxide nanoparticles (SPIONs) have been produced and used as contrast-enhancing agents in magnetic resonance imaging (MRI) for diagnostic use in a wide range of maladies including cardiovascular, neurological disorders, and cancer. The reasons why these SPIONs are attractive for medical purposes are based on their important and unique features. The large surface area of the nanoparticles and their manipulation through an external magnetic field are features that allow their use for carrying a large number of molecules such as biomolecules or drugs. In this scenario, the present work reports on the synthesis and characterization of SPIONs and in vitro MRI experiments to increase their capacity as probes for MRI applications on stem cells therapy. Initially, the SPIONs were prepared through the co-precipitation method using ferrous and ferric chlorides in acidic solution. The SPIONs were coated with two thiolmolecules such as mercaptosuccinic acid (MSA) and cysteine (Cys) (molar ratio SPIONs:ligand = 1:20), leading to the formation of a stable aqueous dispersion of thiolated nanoparticles (SH-SPIONs). The SH-SPIONs were characterized by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), transmission electron microscopy (TEM), and vibrating sample magnetometry (VSM). The results showed that the SH-SPIONs have a mean diameter of 14 nm and display superparamagnetic behavior at room temperature. Preliminary tests of incorporation of SH-SPIONs were evaluated stem cells. The results showed that the thiolated nanoparticles have no toxic effects for stem cells and successfully internalized and enhance the contrast in MRI.

In multisource industrial scenarios (MSIS) coexist NOAA generating activities with other productive sources of airborne particles, such as parallel processes of manufacturing or electrical and diesel machinery. A distinctive characteristic of MSIS is the spatially complex distribution of aerosol sources, as well as their potential differences in dynamics, due to the feasibility of multi-task configuration at a given time. Thus, the background signal is expected to challenge the aerosol analyzers at a probably wide range of concentrations and size distributions, depending of the multisource configuration at a given time. Monitoring and prediction by using statistical analysis of time series captured by on-line particle analyzersin industrial scenarios, have been proven to be feasible in predicting PNC evolution provided a given quality of net signals (difference between signal at source and background). However the analysis and modelling of non-consistent time series, influenced by low levels of SNR (Signal-Noise Ratio) could build a misleading basis for decision making. In this context, this work explores the use of stochastic models based on ARIMA methodology to monitor and predict exposure values (PNC). The study was carried out in a MSIS where an case study focused on the manufacture of perforated tablets of nano-TiO₂ by cold pressing was performed.

The development of nanotechnology has reached a point where it is being widely applied, and numerous nanomaterials and nano-enabled products are handled across a broad range of industrial sectors. Exposure extends beyond occupational settings as products containing nanomaterials are used by different consumer groups.

Despite the knowledge on their toxic effects is growing there is still not OEL for most NMS and therefore the precautionary approach is still used where levels are kept as low as possible Therefore there is a need to assess workers and consumers exposure.

The use of engineered nanoparticles (NP) is more and more widespread in various industrial sectors. The inhalation route of exposure is a matter of concern (adverse effects of air pollution by ultrafine particles and asbestos). No NP biomonitoring recommendations or standards are available so far. The LBM laboratory is currently studying several approaches to develop bioindicators for occupational health applications. As regards exposure indicators, new tools are being implemented to assess potentially inhaled NP in non-invasive respiratory sampling (nasal sampling and exhaled breath condensates (EBC)). Diverse NP analytical characterization methods are used (ICP-MS, dynamic light scattering and electron microscopy coupled to energy-dispersive X-ray analysis). As regards effect indicators, a methodology has been developed to assess a range of 29 cytokines in EBCs (potential respiratory inflammation due to NP exposure). Secondly, collaboration between the LBM laboratory and the EDyp team has allowed the EBC proteome to be characterized by means of an LC-MS/MS process. These projects are expected to facilitate the development of individual NP exposure biomonitoring tools and the analysis of early potential impacts on health. Innovative techniques such as field-flow fractionation combined with ICP-MS and single particle-ICPMS are currently being explored. These tools are directly intended to assist occupational physicians in the identification of exposure situations.

The present work is focused on the measurement of workers exposure to nano-TiO₂ in the life cycle steps of depollutant mortars. It has been done in the framework of the SCAFFOLD project, which aims at the management of potential risks arising from the use of manufactured nanomaterials in construction. Main findings can be summarized as follows: (1) The occupational exposure to nano-TiO₂ is below 0.3 mg/m³ for all measured scenarios. The highest concentrations were measured during the cleaning task (in the nano-TiO₂ manufacturing process) and during the application (spraying) of depollutant coatings on a wall.

It was found a high release of particles above the background in several tasks as expected due to the nature of the activities performed. The maximum concentration was measured during drilling and during adding powder materials (mean total particle concentration up to 5.591E+04 particles/cm³ and 5.69E+04 particles/cm³). However, considering data on total particle concentration released, no striking differences have been observed when tasks have been performed using conventional materials in the sector (control) and when using materials doped with nano-objects.

The implementation in many products of manufactured nanoparticles is growing fast and raises new questions. For this purpose, the CEA - NanoSafety Platform is developing various research topics for health and safety, environment and nanoparticles exposure in professional activities. The containment optimisation for the exposition lowering, then the exposure assessment to nanoparticles is a strategy for safety improvement at workplace and workspace. The lowering step consists in an optimisation of dynamic and static containment at workplace and/or workspace. Generally, the exposure risk due to the presence of nanoparticles substances does not allow modifying the parameters of containment at workplace and/or workspace. Therefore, gaseous or nanoparticulate tracers are used to evaluate performances of containment. Using a tracer allows to modify safely the parameters of the dynamic containment (ventilation, flow, speed) and to study several configurations of static containment. Moreover, a tracer allows simulating accidental or incidental situation. As a result, a safety procedure can be written more easily in order to manage this type of situation. The step of measurement and characterization of aerosols can therefore be used to assess the exposition at workplace and workspace. The case of study, aim of this paper, concerns the potential emission of Lead nanoparticles at the exhaust of a furnace in an epitaxy laboratory. The use of Helium tracer to evaluate the performance of containment is firstly studied. Secondly, the exposure assessment is characterised in accordance with the French guide "recommendations for characterizing potential emissions and exposure to aerosols released from nanomaterials in workplace operations". Thirdly the aerosols are sampled, on several places, using collection membranes to try to detect traces of Lead in air.

This study aims at developing the measurement method of nanoparticle concentration and at getting a representative value of nanoparticle uniform concentration due to chamber ventilation. We conducted a chamber equipped with HEPA filter and control the background nanoparticles concentration by using an adequate ventilation. Then, we used generator to evaluate concentration in the chamber uniformity. We measured background value and source counts at the particle size distribution by SMPS. In addition, we performed numerical analysis with CFD model OpenFoam. As results, we found that there is no aggregate in experimental conditions in this study. Though we confirmed that it is difficult to uniformalise nanoparticle concentration, However we also found simulation results showed higher reproducibility. Therefore, we could assess nanoparticle size distribution and concentration in our chamber at this stage.

Multi-walled carbon nanotubes (MWCNTs) are used as a filler in composites to obtain electrical conductivity, and improve mechanical strength and other properties. However, exposure to MWCNTs may pose health risks because of their size, shape, and insolubility. A quantitative exposure assessment method for CNTs is therefore needed. We have developed a promising carbon analysis method that considers the size distribution of elemental carbon. We conducted exposure assessment according to the lifecycle of CNTs. At the first stage, large quantity of CNTs are handled and exposure to neat CNTs is likely to occur. When large quantity of CNTs are handled, enclosure and automated process are strongly recommended. By applying appropriate measures, CNT concentration can be well controlled. Local exhaust ventilation and less-restrictive enclosures were found to work well during the second stage, which involves handling smaller CNT quantities. At measured sites, MWCNT concentrations were below an occupational exposure level proposed by Nakanishi (i.e., 0.030 mg/m³). This analysis method can also be applied to particles containing MWCNTs. At downstream stages of the lifecycle, neat MWCNTs were not observed and concentrations of embedded MWCNTs were lower than 0.015 mg/m³.

Computational Fluid Dynamics (CFD) codes allow detailed simulation of the flow of gases through fibrous filter media. When the pattern of gas flow between fibers has been established, simulated particles of any desired size can be "injected" into the entering gas stream, and their paths under the influence of aerodynamic drag, Brownian motion and electrostatic forces tracked. Particles either collide with a fiber, or pass through the entire filter medium. They may bounce off the fiber surface, or adhere firmly to the surface or to particles previously captured. Simulated injection of many particles at random locations in the entering stream allows the average probability of capture to be calculated. Many particle properties must be available as parameters for the equations defining the forces on particles in the gas stream, at the moment of contact with a fiber, and after contact. Accurate values for all properties are needed, not only for predicting particle capture in actual service, but also to validate models for media geometries and computational procedures used in CFD. We present a survey of existing literature on the properties influencing nanoparticle dynamics and adhesion.

The objective of the project was to develop a method allowing for detailed characterization of welding particles including particle number concentration, size distribution, surface chemistry and chemical composition of individual particles, as well as metal concentration of various welding fumes in personal exposure samples using regular sampling equipment. A sample strategy was developed to evaluate the variation of the collection methods on mass concentration. Samples were collected with various samplers and filters at two different locations using our collection system. The first location was using a robotic welding system while the second was manual welding. Collected samples were analysed for mass concentration using gravimetryand metal concentration using ICP/OES. More advanced analysis was performed on selected filters using X-Ray Photoelectron Spectroscopy to determine surface composition of the particles, and X-Ray Diffraction to determine chemical composition of the fumes. Results showed that the robotic system had a lot of variation in space when the collection system was located close to the weld. Collection efficiency was found to be quite variable depending upon the type of filter. As well, metal concentrations in blank filters were dependent upon the type of filter with MCE presenting with the highest blank values. Results obtained with the XRD and XPS systems showed that it was possible to analyse a small of powdered welding fume sample but results on filters were not conclusive.

The aim of this study was to design a laboratory size exposure chamber for the testing of samplers used to collect personal exposure samples for nanoparticles. A polyethylene cylindrical container with a diameter of 42 cm and height of 60 cm was used as the testing chamber. The chamber was divided into 2 parts by an aluminium honey comb. Particles generated using a 1 jet Collison nebulizer (BGI) operating at a flow rate of 4L/min were inserted into the chamber via a tube located near to the top of the chamber. A heater was inserted just after the nebulizer to avoid condensation of water in the tubing, and dilution air, running at 10L/min was inserted just after the heater. As particle charge can dramatically affect sampling a particle neutralizer was attached to the generation system so as to neutralize the particles before they enter the chamber. A diffusion dryer was used to remove any water from the air stream prior to enter the chamber. A fan was used to mix and distribute the generated particles. After generation and mixing, the particles passed through the aluminium honeycomb which is essential to eliminate any turbulent or unwanted air flow. Six sampling ports along with a pressure gauge were placed on the walls 15 cm from the bottom of the chamber. The pressure gauge was added to ensure the desired pressure is achieved during sampling. The sampling ports allowed for the connection of five samplers and sampling pumps as well as the connection of an ultrafine particle counter. The exposure chamber was developed to assess various samplers for carbon nanotubes and cellulose nanocrystals. Results showed that the chamber was working properly and that mixing was sufficiently uniform to test samplers.

Electrical mobility classification (EC) followed by Condensation Particle Counter (CPC) detection is the technique combined in Scanning Mobility Particle Sizers(SMPS) to retrieve nanoparticle size distributions in the range from 2.5 nm to 1 μm. The detectable size range of SMPS systems can be extended by the addition of an Optical Particle Sizer(OPS) that covers larger sizes from 300 nm to 10 μm. This optical sizing method reports an optical equivalent diameter, which is often different from the electrical mobility diameter measured by the standard SMPS technique. Multi-Instrument Manager (MIM^TM) software developed by TSI incorporates algorithms that facilitate merging SMPS data sets with data based on optical equivalent diameter to compile single, wide-range size distributions. Here we present MIM 2.0, the next-generation of the data merging tool that offers many advanced features for data merging and post-processing. MIM 2.0 allows direct data acquisition with OPS and NanoScan SMPS instruments to retrieve real-time particle size distributions from 10 nm to 10 μm, which we show in a case study at a fireplace. The merged data can be adjusted using one of the merging options, which automatically determines an overall aerosol effective refractive index. As a result an indirect and average characterization of aerosol optical and shape properties is possible. The merging tool allows several pre-settings, data averaging and adjustments, as well as the export of data sets and fitted graphs. MIM 2.0 also features several post-processing options for SMPS data and differences can be visualized in a multi-peak sample over a narrow size range.

The ability of thermal carbon analysis to determine CNTs was evaluated in the presence of a polymer (Polystyrene, PS). Samples placed in an Au (Pt) foil boat were measured using a thermal-carbon analyzer, and the results were compared with gravimetric measurements of sample masses obtained using an ultra-microbalance. First, debris from the polymer without CNTs (i.e., PS debris) was analyzed. The amount of PS debris detected in the organic carbon (OC) fraction was found to be in good agreement with the gravimetrically measured mass of the PS debris, while the amount of pyrolyticallygenerated carbon soot detected in the elemental carbon (EC) fraction was negligible. Next, single-wall CNT (AIST/TASC Super-Growth) powder was analyzed, and the amount of the CNT powder detected in the EC fraction was found to be 95-96% of the gravimetrically measured mass of the CNT powder. Subsequently, a mixture of the PS debris and the CNT powder was analyzed, and the amounts of detected OC and EC were found to be comparable to the gravimetrically measured masses of the PS debris and the CNT powder, respectively. Finally, debris from 5 wt% CNT-PS composites was analyzed, and amounts of OC and EC detected were found to be approximately comparable to the estimated masses of the PS and the CNTs in the debris of CNT-PS composite, respectively. The results therefore indicate thermal carbon analysis is capable of determining CNTs in the presence of PS.

It has been demonstrated the feasibly of using PLGA nanoparticles to promote the encapsulation of novel anti-diabetic sulphonylhydrazone and antitumor N-acylhydrazone derivatives. The motivation is to further demonstrate the possibility of long-term release of anti-diabetic as well as higher accumulation of the antitumor derivative by using the nanotechnology-based production. The produced nanoparticles were obtained by the nanoprecipitation method, which revealed to be effective in the encapsulation of the bioactive compounds. The determined sizes were in the range of ∼100 nm, which are supposed to be suitable for both potential applications. The preliminary experimental data demonstrated the formation of stable nanosystems and further experiments are underway in order to determine the loading content, encapsulation efficiency and release profile of the hydrophobic bioactive compounds.

The exposure to nanomaterials can yield changes in the mineral composition of tissues which may have long term health repercussions. In this study, the changes in mineral composition of rat lungs, exposed to a nanoaerosol of silicon carbide (SiC), has been studied by means of global and local ion beam probes with the Particle-Induced X-ray Emission (PIXE) technique, measuring the whole lung contents and selected areas where SiC was found, respectively. It was found that from a global perspective there is a small decrease in the mineral contents (phosphorous, sulphur, chlorine and potassium) of the lung except for Ca, while locally these mineral contents tend fluctuate.

In order to study a scenario of acute high concentration exposure via the pulmonary pathway of silicon carbide and titanium carbide nanoparticles, female Wistar rats were administered by intratracheal instillation doses of 0.5 and 5 mg/rat of each nanomaterial. Inflammatory parameters were studied: protein concentration, lactate dehydrogenase activity, total cell count and differentiated cell count (macrophages, neutrophils, oesonophils, lymphocytes). The genotoxicity potential was assessed by the formation of micronuclei from pneumocytes type II. It was found that silicon carbide nanoparticles induce an inflammatory response and a dose dependent genotoxicity, although the genotoxicity levels are comparably lower to the inflammatory response.

Recently, it has been proven that manganese from inhaled particles of manganese compounds can accumulate in the internal organs of laboratory animals. Nevertheless, there were only a few researches dealing with changes in body morphology induced by inhalation of these particles, even though results of some studies indicate existence of such changes. The aim of our research was to assess the effect of inhaled manganese oxides nanoparticles on weight of internal organs. For this purpose a long-term inhalation experiment on laboratory mice was performed, during which the mice were exposed to MnO.Mn₂O₃ nanoparticles in concentration 2 × 10⁶ particles/cm³ for 17 weeks, 24 hours a day, 7 days a week. Manganese oxides nanoparticles were synthesized continuously via aerosol route in a hot wall tube flow reactor using thermal decomposition of metal organic precursor manganese(II)acetylacetonate in the flow tube reactor at temperature 750 °C in the presence of 30 vol% of oxygen. It was proven that inhaled nanoparticles can influence the weight of internal organs of mice. Moreover, it was discovered that the resulting change in weight of selected organs is disproportional. The mice from the experimental group had statistically significantly lighter kidneys, liver and spleen and heavier pancreas compared to the mice from the control group.

The graphene sample GO:Single-layer graphene oxide, purity 99%, thickness 0.7-1.2 nm (AFM); ∼300-800nm X&Y dimensions is the standard size <450 nm & 1-20 μm lateral dimensions. Cheap Tubes Inc., Bratleboro, USA was selected for our study. Exhaustive characterization of GO was afforded. It exhibited thermal stability over 60^oC and it was suspended in deionized water after ultrasonication(1 mg/mL) (stable 10 days). All the biological fluids used in the different assays were used as control of the colloidal suspension stability. Then, all the studies were carried out within that stability period. The cytotoxicity assays were carried out by the Alamar Blue (Resazurin) reduction, MTT and flow cytometry assays in mouse embryonic fibroblast cells (3T3), human keratinocytes (HaCaT), colorectal cancer cells (Caco-2/HCT 116), Lewis lung cancer cells (3LL), acute myeloid leukemia cells (KG-1, Jurkat, Kasumi-1) and chronic myeloid leukemia cells (K562, Lucena) and no significant toxicity was found after exposition to 0.1-100 μg/mL for 24 and 48 h. Breast cancer cells, MCF-7, showed a 20% reduction on cell viability at 24 and 48 h. No cytotoxicity were found in lymphocytes, Chinese hamster ovary cells (CHO) and human macrophage cell line (U937) at 0.1-50 μg/mL, but 30-50% survival inhibition was observed at 100 μg/mL. A dose-dependent increase in apoptosis was observed in some cells (Kasumi-1, Jurkatand K562 cells). In the case of CHO and 3T3 cells, greater levels of necrosis with increasing concentrations of GO (>50 μg/mL) were observed. Genotoxic study using the Comet assay showed slight DNA damage in lymphocytes at all concentrations tested, while more significant effects was observed in CHO cells. Econanotoxicity was carried out by lethality assays in the nematode Caenorhabditis elegans, d in the freshwater coelenterate Hydra, Daphania amd in Shrimp with no signs of toxicity at concentrations varying from 0.1-100 μg/mL of GO. However, death and disintegration of Hydra was observed after exposition to 100 μg/mL for 72 h. In in vivo studies, no changes in biochemical parameters of Fischer 344 rats were observed after the i.p. administration of GO. Some black agglomerates were found in the intraperitoneal cavity of rats injected with GO. However, in Fisher 344 rats-bearing prostate tumors, treatment with GO (up to 100 μg/mL) negatively affected the hepatic parameters, whilst in the renal ones, an improvement was observed. Studies are in progress to understand the mechanisms involved in the uptake of GO by RES. GO appears as a potential non-toxic in vitro and in vivo assays at the concentrations used in this interlab experiments.

Chagas disease, also known as American trypanosomiasis, is a potentially life-threatening illness caused by the protozoan parasite, Trypanosoma cruzi (T. cruzi), and the disease remains a major health problem in many Latin American countries. Several papers report that the killing of the parasite is dependent on the production of nitric oxide (NO). The endogenous free radical NO is an important cellular signalling molecule that plays a key role in the defense against pathogens, including T. cruzi. As T. cruzi is able to compromise host macrophages decreasing endogenous NO production, the administration of exogenous NO donors represents an interesting strategy to combat Chagas disease. Thus, the aims of this study were to prepare and evaluate the antimicrobial activity of NO-releasing polymeric nanoparticles against T. cruzi. Biocompatible polymeric nanoparticles composed of chitosan/sodium tripolyphosphate(TPP) were prepared and used to encapsulate mercaptosuccinic acid (MSA), which is a thiol-containing molecule. Nitrosation of free thiols (SH) groups of MSA were performed by the addition of equimolar amount of sodium nitrite (NaNO₂), leading to the formation of S-nitroso-MSA-containing nanoparticles. These polymeric nanoparticles act as spontaneous NO donors, with free NO release. The results show the formation of nanoparticles with average hydrodynamic diameter ranging from 270 to 500 nm, average of polydispersity index of 0.35, and encapsulation efficiency in the range of 99%. The NO release kinetics from the S-nitroso-MSA-containing nanoparticles showed sustained and controlled NO release over several hours. The microbicidal activity of S-nitroso-MSA-containing nanoparticles was evaluated by incubating NO-releasing nanoparticles (200 - 600 μg/mL) with replicative and non-infective epimastigote, and non-replicative and infective trypomastigote forms of T. cruzi. In addition, a significant decrease in the percentage of macrophage-infected (with amastigotes) and NO-releasing nanoparticle-treated cells was observed. Taken together, our results reveal a potent toxic effect of NO-releasing polymeric nanoparticles against different life cycle forms of T. cruzi, indicating that the encapsulation of the NO donor S-nitroso-MSA represents an interesting approach to combat and to prevent Chagas disease.

It was prepared the graphene oxide (GO) sheets by suspension of GO in ultrapure deionized water or in Pluronic F-68 using a ultrasonicator bath. Total characterization of GO sheets was carried out. The results on suspension of GO in water showed excellent growth and cell adhesion. GO/Pluronic F-68 platform for the growth and adhesion of adipose-derived stem cells (ASCs) that exhibits excellent properties for these processes. GO in water suspension exhibited an inhibition of the cell growth over 5 μg/mL In vivo study with GO suspended in water (100 μg/mL) on Fisher 344 rats via i.p. administration showed low toxicity. Despite GO particle accumulates in the intraperitoneal cavity, this fact did not interfere with the final absorption of GO. The AST (aspartate aminotransferase) and ALT (alanine aminotransferase) levels (liver function) did not differ statistically in all experimental groups. Also, creatinine and urea levels (renal function) did not differ statistically in all experimental groups. Taking together, the data suggest the great potential of graphene oxide sheets as platform to ACSs, as well as, new material for treatment several urological diseases.

The present work is focused on the synthesis, characterization and cytotoxic evaluation of superparamagnetic iron oxide nanoparticles (SPIONs). SPIONs have been proposed for an increasing number of biomedical applications, such as drug-delivery. To this end, toxicological studies of their potential effects in biological systems must be better evaluated. The aim of this study was to examine the in vitro cytotoxicity of thiolated (SH) and S-nitrosated (S-NO) SPIONs in cancer cell lines. SPIONs were prepared by the coprecipitation method using ferrous and ferric chlorides in aqueous solution. The nanoparticles (Fe₃O₄) were coated with thiol containing molecule cysteine (Cys) (molar ratio SPIONs:ligand = 1:20), leading to the formation of an aqueous dispersion of thiolated nanoparticles (SH- SPIONs). These particles were characterized by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM). The results obtained showed that Cys-SPIONs have a mean diameter of 14 nm at solid state and present super paramagnetic behavior at room temperature. Thiol groups on the surface of the nanoparticles were nitrosated through the addition of sodium nitrite leading to the formation of S-NOCys-SPIONs (S-nitrosated-Cys-SPIONs), which act as spontaneous nitric oxide (NO) donor). The cytotoxicity of thiolated and S-nitrosated nanoparticles was evaluated in acute T cell leukemia (Jurkat cell line) and Lewis lung carcinoma (3LL) cells. The results showed that at low concentrations thiolated (Cys) and S- nitrosated (S-NOCyst) SPIONs display low cytotoxicity in both cell types. However, at higher concentrations, Cys-SPIONs exhibited cytotoxic effects, whereas S-NOCys-SPIONs protected them, and also promoted cell proliferation.

In recent years, production of nanoparticles is increased and thus grows our contact with them too. Question of safety is closely related to the issue of use nanoparticles. There are a number of tests that monitor the viability, ROS production, the effect on the DNA and cell cycle, however, rarely encountered studies on stress in the cells after contact with nanoparticles. Heat shock proteins (HSP) are among the substances that can be used for monitoring stress in cells. HSP are structures with a chaperone activity. They are evolutionarily very old, conservative and they are found with a high degree of homology in prokaryotes and eukaryotes including humans. They exist at low concentrations under physiological conditions, while in the denaturing conditions e.g. high or low temperature, radiation, exposure to chemicals, heavy metals, or nanoparticles their expression is changed. HSPs are involved in maintaining homeostasis in the cell that the denatured protein conformations allow recovery to the original stage. One of the most common proteins from HSP family is Hsp70 - protein with a molecular weight of 70 kDa. The level of Hsp70 in a cell after exposure to the stress changes depending on the stress level to which the cell is exposed to and a time period during which lasted stressful conditions. Our research monitors stress levels of cells manifesting by Hsp70 production after contact with silver nanoparticles. Nanoparticles show different toxicity towards different types of target cells, which is reflected in the values of IC₅₀ - concentration that kills 50% tested cells. Concentration of test substance toxic to one cell type may be innocuous to cells of another type. IC₅₀ obtained from the MTT assay provides a suitable default data and if multiples of IC₅₀ values are used, we can compare and generalize. Studies can be used to compare stress levels in cells that show different sensitivity to the tested nanoparticles compared with cells under optimal growth conditions. The study was done on two types of mouse fibroblasts NIH-3T3 and L929. While NIH-3T3 cells exhibit stress response proportional to the concentration of silver nanoparticles, for L929 cells this was not observed.

One of the major problems in cancer therapies is the high occurrence of side effects intrinsic of anticancer drugs. Doxorrubicin is a conventional anticancer molecule used to treat a wide range of cancer, such as breast, ovarian and prostate. However, its use is associated with a number of side effects like multidrug resistance and cardiotoxicity. The association with nanomaterials has been considered in the past decade to overcome the high toxicity of these drugs. In this context, mesoporous silica nanoparticles are great candidates to be used as carriers once they are very biocompatible. Taking into account the combination of nanoparticles and doxorrubicin, we treated rats with chemically induced prostate cancer with systems based on mesoporous silica nanoparticles and a thermoreversible block copolymer (Pluronic F-127) containing doxorrubicin. Preliminary results show a possible improvement in tumor conditions proportional to the concentration of the nanoparticles, opening a perspective to use mesoporous silica nanoparticles as carrier for doxorrubicin in prostate cancer treatment.

Nowadays, there are several commercially available products containing nanostructured materials. Meanwhile, despite the many benefits that can be obtained from nanotechnology, it is still necessary to understand the mechanisms in which nanomaterials interact with the environment, and to obtain information concerning their possible toxic effects. In agriculture, nanotechnology has been used in different applications, such as nanosensors to detect pathogens, nanoparticles as controlled release systems for pesticides, and biofilms to deliver nutrients to plants and to protect food products against degradation. Moreover, plants can be used as models to study the toxicity of nanoparticles. Indeed, phytotoxicity assays are required to identify possible negative effects of nanostructured systems, prior to their implementation in agriculture. Nitric oxide (NO) plays a key role in plant growth and defense, and recently, several papers described the beneficial effects due to application of exogenous NO donors in plants. The tripeptide glutathione (GSH) is an important anti-oxidant molecule and is the precursor of the NO donor, S-nitrosoglutathione (GSNO). In this context, the present work investigates the effects of different concentrations of alginate/chitosan nanoparticles, containing either GSH or GSNO, on the development of two test species (Zea mays and Glycine sp.). The results showed that the alginate/chitosan nanoparticles present a size average range from 300 to 550 nm with a polydispersity index of 0.35, and encapsulation efficiency of GSH between 45 - 56%. The NO release kinetics from the alginate/chitosan nanoparticles containing GSNO showed sustained and controlled NO release over several hours. Plant assays showed that at the concentrations tested (1, 5 and 10 mM of GSH or GSNO), polymeric nanoparticles showed no significant inhibitory effects on the development of the species Zea mays and Glycine sp., considering the variables shoot height, root length, and dry mass. Therefore, these nanoparticles seem to have promissing uses in agriculture, and might be potencially used as controlled release systems applied by the foliar route.

Hazard studies of "as-produced" nanomaterials are increasingly available, yet a critical gap exists in exposure science that may impede safe development of nanomaterials. The gap is that we do not understand what is actually released because nanomaterials can change when released in ways that are not understood. We also generally do not have methods capable of quantitatively measuring what is released to support dose assessment. This review presents a case study of multi-walled carbon nanotubes (MWCNTs) for the measurement challenge to bridge this gap. As the use and value of MWCNTs increases, methods to measure what is released in ways relevant to risk evaluation are critically needed if products containing these materials are to be economically, environmentally, and socially sustainable. This review draws on the input of over 50 experts engaged in a program of workshops and technical report writing to address the release of MWCNTs from nanocomposite materials across their life cycle. The expert analyses reveals that new and sophisticated methods are required to measure and assess MWCNT exposures for realistic exposure scenarios. Furthermore, method requirements vary with the materials and conditions of release across life cycle stages of products. While review shows that the likelihood of significant release of MWCNTs appears to be low for many stages of composite life cycle, measurement methods are needed so that exposures from MWCNT-composites are understood and managed. In addition, there is an immediate need to refocus attention from study of "as-produced" nanomaterials to coordinated research on actual release scenarios.

Nanomaterials are one of the promising technologies of this century. The Project on Emerging Nanotechnologies [1] reports more than 1600 consumer products based on nanotechnology that are currently on the market and advantages link to the reinforcement of polymeric materials using nano-fillers are not to demonstrate anymore. However, the concerns about safety and its consumer perception can slow down the acceptance of nanocomposites. Indeed, during its life-cycle, a nanotechnology-based product can release nano-sized particles exposing workers, consumers and environment and the risk involved in the use and disposal of such particles is not well known. The current legislation concerning chemicals and environment protection doesn't explicitly cover nanomaterials and changes undergone by nanoparticles during the products' life cycle. Also, the possible physio-chemical changes that the nanoparticles may undergo during its life cycle are unknown. Industries need a standard method to evaluate nanoparticles release during products' life cycle in order to improve the knowledge in nanomaterials risk assessment and the legislation, and to inform customers about the safety of nanomaterials and nanoproducts. This work aims to propose a replicable method in order to assess the release of nanoparticles during the machining of nanocomposites in a controlled environment. For this purpose, a new experimental set-up was implemented and issues observed in previous methods (background noise due to uncontrolled ambient environment and the process itself, unrepeatable machining parameters) were solved. A characterisation and validation of the chamber used is presented in this paper. Also, preliminary testing on drilling of polymer-based nanocomposites (Polyamide-6/Glass Fibre reinforced with nano-SiO₂) manufactured by extrusion and injection moulding were achieved.

The aerosol particles released during the grinding of polystyrene (PS)-based composites with well- and poorly dispersed single-wall carbon nanotubes (CNTs) and multiwall CNTs were measured using real-time aerosol measuring instruments. Increases in the concentration of aerosol particles were recorded during the grinding of the samples. However, similar increases were observed even when CNT-free polystyrene was ground. Electron microscopic analysis of the released particles revealed that particles with protruding CNTs were observed for the well-dispersed CNT-PS composites, but free-standing CNTs were not found. On the other hand, particles like agglomerated CNTs were found for the poorly dispersed CNT-PS composites.

Currently, the characterisation of particle release from nanostructured powders is realised based on different release scenarios, which were typically designed to describe real powder handling processes. Most of these scenarios base on short term stress situations (e.g. drop down, mixing) and are thus accompanied with dynamic changes of the particle number concentration, which complicates both the process characterisation and the release characterisation. In this study, two continuous release processes for weak and intense dispersion of powder agglomerates were analysed based on considerations regarding energetic and instrumental impact.

Parallel to the increased use of engineered nanoparticles (ENP) in the formulation of commercial products or in medicine, numerous health & safety agencies have recommended the application of the precautionary principle to handle ENP; namely, the recommendation to use protective gloves against chemicals. However, recent studies reveal the penetration of titanium dioxide nanoparticles through nitrile rubber protective gloves in conditions simulating occupational use. This project is designed to understand the links between the penetration of gold nanoparticles (nAu) through nitrile rubber protective gloves and the mechanical and physical behaviour of the elastomer material subjected to conditions simulating occupational use (i.e., mechanical deformations (MD) and sweat). Preliminary analyses show that nAu suspensions penetrate selected glove materials after exposure to prolonged (3 hours) dynamic deformations. Significant morphological changes are observed on the outer surface of the glove sample; namely, the number and the surface of the micropores on the surface increase. Moreover, nitrile rubber protective gloves are also shown to be sensitive to the action of nAu suspension and to the action of the saline solution used to simulate sweat (swelling).

This research, funded by the Institution of Occupational Safety and Health in the United Kingdom, has used a combination of literature review, web searching and unstructured interviews with a range of industry professionals to compile a list of products used in construction and the built environment which might contain nanomaterials. Samples of these products have been analysed using Scanning Electron Microscopy and Energy Dispersive X- Ray Spectroscopy to investigate whether nanomaterials are actually present and to what extent. Preliminary results of this testing are presented here. It is concluded that there is a discrepancy between the academic literature and the reality regarding the current application of nanomaterials in the construction industry and the built environment. There are also inaccuracies and deficiencies in the information provided by manufacturers which makes it difficult to accurately assess the location and application of nanomaterials within the industry. Further testing is planned to evaluate the risk of nanoparticle release from nano-enabled building products at their end of life by reproducing common demolition and recycling processes such as crushing, grinding, burning and melting. Results of this will form the basis of practical guidance for the construction, demolition and recycling industries to help them identify where particular protection or control measures may be appropriate as well as providing reassurance where no additional action is required.

Nanotechnology can contribute to the development of innovative applications in the agriculture, food and feed sector by e.g. enabling improved delivery of nutrients or increased efficacy of agrichemicals. It is expected that applications will increase in the near future and may therefore become a relevant source of human exposure to nanomaterials (NM). To gain more up-to date information, RIKILT and the Joint Research Centre (JRC) were commissioned by the European Food Safety Authority (EFSA) to prepare an inventory of currently used and reasonably foreseen applications of NM in agriculture and food/feed production and carried out a review of regulatory aspects concerning NM in both EU and non-EU countries. An analysis of the information records in the inventory shows that nano-encapsulates, silver and titanium dioxide are the most frequent type of NM listed and that food additives and food contact materials are the most frequent types of application. A comparison between marketed applications and those in development indicates a trend from inorganic materials (e.g. silver) towards organic materials (nano-encapsulates, nanocomposites). Applications in novel food, feed additives, biocides and pesticides are currently mostly at a developmental stage. The review of EU and non-EU legislation shows that currently a few EU legal acts incorporate a definition of a nanomaterial and specific provisions for NM, whereas in many non-EU countries a broader approach is applied, which mainly builds on guidance for industry.

Nanotechnology is considered one of the key technologies of the 21^st century within Europe and a Key-Enabling Technology (KET) by Horizon 2020. Standardization has been identified in H2020 as one of the innovation-support measures by bridging the gap between research and the market, and helping the fast and easy transfer of research results to the European and international market. The development of new and improved standards requires high quality technical information, creating a fundamental interdependency between the standardization and research communities. In the frame of project nanoSTAIR (GA 319092), the present paper describes the European scenario on research and standardization in nanotechnology and presents a proposal of a European strategy (nanoSTAIR) to impulse direct "pipelines" between research and standardization. In addition, strategic actions focused on integration of standardization in the R&D projects, from the early stages of the design of a future business (Project Proposal), are also described.

Nanoparticles in workplaces may pose a threat to the health of the workers involved. With the general boom in nanotechnology, an increasing number of workers is potentially exposed, and therefore a comprehensive risk management with respect to nanoparticles appears necessary. One (of many) components of such a risk management is the measurement of personal exposure. Traditional nanoparticle detectors are often cumbersome to use, large, heavy and expensive. We have developed small, reliable and easy to use devices that can be used for routine personal exposure measurement in workplaces.

This paper presents a general overview of the work carried out by European project SCAFFOLD (GA 280535) during its 30 months of life, with special emphasis on risk management component. The research conducted by SCAFFOLD is focused on the European construction sector and considers 5 types of nanomaterials (TiO₂, SiO₂, carbon nanofibres, cellulose nanofibers and nanoclays), 6 construction applications (Depollutant mortars, selfcompacting concretes, coatings, self-cleaning coatings, fire resistant panels and insulation materials) and 26 exposure scenarios, including lab, pilot and industrial scales. The document focuses on the structure, content and operation modes of the Risk Management Toolkit developed by the project to facilitate the implementation of "nano-management" in construction companies. The tool deploys and integrated approach OHSAS 18001 - ISO 31000 and is currently being validated on 5 industrial case studies.

The French national epidemiological surveillance program EpiNano aims at surveying mid- and long-term health effects possibly related with occupational exposure to either carbon nanotubes or titanium dioxide nanoparticles (TiO₂). EpiNano is limited to workers potentially exposed to these nanomaterials including their aggregates and agglomerates. In order to identify those workers during the in-field industrial hygiene visits, a standardized non-instrumental method is necessary especially for epidemiologists and occupational physicians unfamiliar with nanoparticle and nanomaterial exposure metrology. A working group, Quintet ExpoNano, including national experts in nanomaterial metrology and occupational hygiene reviewed available methods, resources and their practice in order to develop a standardized tool for conducting company industrial hygiene visits and collecting necessary information. This tool, entitled "Onsite technical logbook", includes 3 parts: company, workplace, and workstation allowing a detailed description of each task, process and exposure surrounding conditions. This logbook is intended to be completed during the company industrial hygiene visit. Each visit is conducted jointly by an industrial hygienist and an epidemiologist of the program and lasts one or two days depending on the company size. When all collected information is computerized using friendly-using software, it is possible to classify workstations with respect to their potential direct and/or indirect exposure. Workers appointed to workstations classified as concerned with exposure are considered as eligible for EpiNano program and invited to participate. Since January 2014, the Onsite technical logbook has been used in ten company visits. The companies visited were mostly involved in research and development. A total of 53 workstations with potential exposure to nanomaterials were pre-selected and observed: 5 with TiO₂, 16 with single-walled carbon nanotubes, 27 multiwalled carbon nanotubes. Among the tasks observed there were: nanomaterial characterisation analysis (8), weighing (7), synthesis (6), functionalization (5), and transfer (5). The manipulated quantities were usually very small. After analysis of the data gathered in logbooks, 30 workstations have been classified as concerned with exposure to carbon nanotubes or TiO₂. Additional tool validity as well as inter-and intra-evaluator reproducibility studies are ongoing. The first results are promising.

Polymer binder modification with inorganic nanomaterials (NM) could be a potential and efficient solution to control matrix flammability of polymer concrete (PC) materials without sacrificing other important properties. Occupational exposures can occur all along the life cycle of a NM and "nanoproducts" from research through scale-up, product development, manufacturing, and end of life. The main objective of the present study is to analyse and compare different qualitative risk assessment methods during the production of polymer mortars (PM) with NM. The laboratory scale production process was divided in 3 main phases (pre-production, production and post-production), which allow testing the assessment methods in different situations. The risk assessment involved in the manufacturing process of PM was made by using the qualitative analyses based on: French Agency for Food, Environmental and Occupational Health & Safety method (ANSES); Control Banding Nanotool (CB Nanotool); Ecole Polytechnique Fédérale de Lausanne method (EPFL); Guidance working safely with nanomaterials and nanoproducts (GWSNN); Istituto Superiore per la Prevenzione e la Sicurezza del Lavoro, Italy method (ISPESL); Precautionary Matrix for Synthetic Nanomaterials (PMSN); and Stoffenmanager Nano. It was verified that the different methods applied also produce different final results. In phases 1 and 3 the risk assessment tends to be classified as medium-high risk, while for phase 2 the more common result is medium level. It is necessary to improve the use of qualitative methods by defining narrow criteria for the methods selection for each assessed situation, bearing in mind that the uncertainties are also a relevant factor when dealing with the risk related to nanotechnologies field.

This paper examines one aspect of innovation dynamics for nanoresponsible development: the links between regulation and innovation dynamics. It focuses on the case of the French Code de l'environnement, Articles L. 523-1 to L. 523-3. Articles L. 523-1 to L. 523-3 of the French environment code provide for the obligation to declare the quantities and uses of substances at nanoscale produced, distributed or imported in France. This procedure is intended to improve knowledge of these substances and their uses as well as of their markets and volumes sold, to ensure traceability and to collect available information on their toxicological and ecotoxicological properties. The paper builds on recent work on the emergence of a regulatory framework for nanotechnologies to take stock of the current situation in France, in the EU and globally and to explore how this specific law package may influence innovation and the shaping of new markets for nanobased materials. The study shows that nano-regulation does have an impact on innovation. However, the impact is not the same with EU regulation and with French regulation, and while EU regulation seems to create a favourable context for innovation, French regulation seems to do the opposite. With this study we hope to bring new perspectives to the field of the strategic management of innovation, and also to shed some light on the roles and challenges of institutions to facilitate nanoresponsible development.

Scientific literature suggests that exposure to nanoparticles (NPs) might be associated with adverse health effects. A well-developed human risk assessment (HRA) that applies to NPs has never been established and optimized–until now. Furthermore, no government regulations are in place that establish what is considered to be an adequate and secure level of exposure and supported by a strong scientific approach for nanotechnology. It is important to implement the HRA to ensure that workers producing NPs, users of NPs and the general population are protected from deleterious issues related to NPs. In this work, a methodology is described based on the HRA. An effort is required during synthesis before the commercialization phase to evaluate the results of a systematic and rigorous assessment because this could significantly reduce the health risks of those exposed to NPs, including workers and the population.