Limitations and information needs for engineered nanomaterial-specific exposure estimation and scenarios: recommendations for improved reporting practices

The aim of this paper is to describe the process and challenges in building exposure scenarios for engineered nanomaterials (ENM), using an exposure scenario format similar to that used for the European Chemicals regulation (REACH). Over 60 exposure scenarios were developed based on information from publicly available sources (literature, books, and reports), publicly available exposure estimation models, occupational sampling campaign data from partnering institutions, and industrial partners regarding their own facilities. The primary focus was on carbon-based nanomaterials, nano-silver (nano-Ag) and nano-titanium dioxide (nano-TiO2), and included occupational and consumer uses of these materials with consideration of the associated environmental release. The process of building exposure scenarios illustrated the availability and limitations of existing information and exposure assessment tools for characterizing exposure to ENM, particularly as it relates to risk assessment. This article describes the gaps in the information reviewed, recommends future areas of ENM exposure research, and proposes types of information that should, at a minimum, be included when reporting the results of such research, so that the information is useful in a wider context.

Mechanisms of interaction of therapeutic nanoparticles with cells

Nanoparticles (NPs) have gained a lot of interest in recent years due to their huge potential for applications in industry and medicine. Their unique properties offer a large number of attractive possibilities in the biomedical field, providing innovative tools for diagnosis of diseases and for novel therapies. Nevertheless, a deep understanding of their interactions with living tissues and the knowledge about their possible effects in the human body are necessary for the safe use of nanoparticulate formulations. The aim of this PhD project was to study in detail the interactions of therapeutic NPs with living cells, including cellular uptake and release, cellular localization and transport across the cell layers. Moreover, the effects of NPs on the cellular metabolic processes were determined using adapted in vitro assays. We evaluated the biological effect of several NPs potentially used in the biomedical field, including titanium dioxide (Ti02) NPs, 2-sized fluorescent silica NPs, ultrasmall superparamagnetic iron oxide (USPIO) NPs, either uncoated or coated with oleic acid or with polyvinylamine (aminoPVA) and poly(lactic-co-glycolic acid) - polyethylene-oxide (PLGA-PEO) NPs. We have found that the NPs were internalized by the cells, depending on their size, chemical composition, surface coating and also depending on the cell line considered. The uptake of aminoPVA-coated USPIO NPs by endothelial cells was enhanced in the presence of an external magnetic field. None of the tested USPIO NPs and silica NPs was transported across confluent kidney cell layers or brain endothelial cell layers, even in the presence of a magnetic field. However, in an original endothelium-glioblastoma barrier model which was developed, uncoated USPIO NPs were directly transferred from endothelial cells to glioblastoma cells. Following uptake, Ti02 NPs and uncoated USPIO NPs were released by the kidney cells, but not by the endothelial cells. Furthermore, these NPs induced an oxidative stress and autophagy in brain endothelial cells, possibly associated with their enhanced agglomeration in cell medium. A significant DNA damage was found in brain endothelial cells after their exposure to TiO2NPs. Altogether these results extend the existing knowledge about the effects of NPs on living cells with regard to their physicochemical characteristics and provide interesting tools for further investigation. The development of the in vitro toxicological assays with a special consideration for risk evaluation aims to reduce the use of animal experiments. -Les nanoparticules (NPs) présentent beaucoup d'intérêt dans le domaine biomédical et industriel. Leurs propriétés uniques offrent un grand nombre de possibilités de solutions innovantes pour le diagnostique et la thérapie. Cependant, pour un usage sûr des NPs il est nécessaire d'acquérir une connaissance approfondie des mécanismes d'interactions des NPs avec les tissus vivants et de leur effets sur le corps humain. Le but de ce projet de thèse était d'étudier en détail les mécanismes d'interactions de NPs thérapeutiques avec des cellules vivantes, en particulier les mécanismes d'internalisation cellulaire et leur subséquente sécrétion par les cellules, leur localisation cellulaire, leur transport à travers des couches cellulaires, et l'évaluation des effets de NPs sur le métabolisme cellulaire, en adaptant les méthodes existante d'évaluation cyto-toxico logique s in vitro. Pour ces expériences, les effets biologiques de nanoparticules d'intérêt thérapeutique, telles que des NPs d'oxyde de titane (TiO2), des NPs fluorescents de silicate de 2 tailles différentes, des NPs, d'oxyde de fer super-para-magnétiques ultra-petites (USPIO), soit non- enrobées soit enrobées d'acide oléique ou de polyvinylamine (aminoPVA), et des NPs d'acide poly(lactique-co-glycolique)-polyethylene-oxide (PLGA-PEO) ont été évalués. Les résultats ont démontré que les NPs sont internalisées par les cellules en fonction de leur taille, composition chimique, enrobage de surface, et également du type de cellules utilisées. L'internalisation cellulaire des USPIO NPs a été augmentée en présence d'un aimant externe. Aucune des NPs de fer et de silicate n'a été transportée à travers des couches de cellules épithéliales du rein ou endothéliales du cerveau, même en présence d'un aimant. Cependant, en développant un modèle original de barrière endothélium-glioblastome, un transfert direct de NPs d'oxyde de fer de cellule endothéliale à cellule de glioblastome a été démontré. A la suite de leur internalisation les NPs d'oxyde de fer et de titane sont relâchées par des cellules épithéliales du rein, mais pas des cellules endothéliales du cerveau. Dans les cellules endothéliales du cerveau ces NPs induisent en fonction de leur état d'agglomération un stress oxydatif et des mécanismes d'autophagie, ainsi que des dommages à l'ADN des cellules exposées aux NPs d'oxyde de titane. En conclusion, les résultats obtenus élargissent les connaissances sur les effets exercés par des NPs sur des cellules vivantes et ont permis de développer les outils expérimentaux pour étudier ces effets in vitro, réduisant ainsi le recours à des expériences sur animaux.

Aqueous photocatalytic oxidation of steroid estrogens

Concerns have increased regarding the detection of endocrine-disrupting compounds in the effluents of sewage treatment plants (STPs). These compounds are able to disrupt normal function of the endocrine system of living organisms even at trace concentrations. Natural and synthetic steroid estrogens (SEs) are believed to be responsible for the majority of the endocrine-disrupting effects. Municipal sewage, the main source of SEs in the environment, is a complex mixture of a wide range of pollutants at concentrations much higher than those of SEs. Low concentrations of SEs in the presence of copollutants thus make their removal problematic. The main objectives of the present work were to study the potential of photocatalytic oxidation (PCO) to effectively treat SE-containing aqueous solutions and to identify the optimum conditions for such treatment. The results showed that SEs can be effectively degraded photocatalytically. Due to the adsorption properties of SEs on the TiO2 photocatalyst surface alkaline medium was found to be beneficial for SE oxidation despite the presence of co-pollutants in concentrations characteristic for the sanitary fraction of municipal sewage. The potential of PCO to selectively oxidise SEs was examined in the presence of copollutants of the sanitary fraction of sewage - urea, saccharose and human urine. The impact of ethanol, often used as a solvent in the preparation of SE stock solutions, was also studied and the results indicated the need to use organic solvent-free solutions for the study of SE behaviour. Photocatalytic oxidation of SEs appeared to be indifferent towards the presence of urea in concentrations commonly found in domestic sewage. The effect of other co-pollutants under consideration was far weaker than could be expected from their concentrations, which are from one hundred to a few thousands times higher than those of the SEs. Although higher concentrations can dramatically slow down the PCO of SEs, realistic concentrations of co-pollutants characteristic for the sanitary fraction of domestic sewage allowed selective removal of SEs. This indicates the potential of PCO to be a selective oxidation method for SE removal from the separate sanitary fraction of municipal sewage.

Photocatalytic oxidation of humic substances and lignins in aqueous solutions

Tässä tutkimuksessa tarkastellaan kahden yleisen, veden ympäristökuormitusta aiheuttavan kemikaaliryhmän, ligniinin ja humusaineiden, fotokatalyyttistahapetusta (photocatalytic oxidation, PCO) vesiliuoksessa. Fotokatalyyttina käytettiin titaanidioksidia, jota säteilytettiin ultraviolettivalolla. Työssä selvitettiin useiden eri olosuhdeparametrien vaikutusta fotokatalyysiin. Tutkittavia parametreja olivat mm. kontaminanttien alkukonsentraatio, pH, vetyperoksidilisäys, rauta-ionien lisäys, fotokatalyysimenetelmä, fotokatalyytin pintakonsentraatioja titaanidioksidin määrä lasisissa mikropartikkeleissa. Ultraviolettivalon lähteinä käytettiin sekä keinovaloa että auringonvaloa. Katalyytin kantoaineena käytettiin huokoisia lasisia mikropartikkeleita, joiden pintaan kiinnittynyt titaanidioksidi pystyi hyvin vähentämään kontaminanttien määrää vedessä. Fotokatalyysin tehokkuus kasvoi humusaine- ja ligniinikonsentraatioiden kasvaessa. Korkeimmat hapetustehokkuudet kumallakin kontaminantilla saavutettiin neutraaleissa jalievästi emäksisissä olosuhteissa huolimatta siitä, että paras adsorboituminen tapahtui happamissa olosuhteissa. Tämän perusteella voidaan olettaa, että humusaineiden ja ligniinin hapetus tapahtuu pääosin radikaalimekanismilla. Vetyperoksidin lisääminen humusaineliuokseen lisäsi hapettumisnopeutta, vaikka näennäinen hapetustehokkuus ei muuttunut. Tämän perusteella vetyperoksidi hapetti myös humusaineita referenssinäytteessä. Ligniinin fotokatalyyttinen hapettuminen parani vetyperoksidilisäyksellä happamissa olosuhteissa johtuen lisääntyneestä OH-radikaalien muodostumisesta. Ligniini ei hapettunut vetyperoksidilla, jos fotokatalyyttiä ei¿ollut läsnä. Rauta-ionit eivät lisänneet humushappojen fotokatalyyttistähapettumista, mutta Fe2+-ionien lisäys aina konsentraatioon 0.05 mM johti ligniinin hapettumistehokkuuden voimakkaaseen kasvuun. Rauta-ionikonsentraation kasvattaminen edelleen johti ligniinin hapetustehokkuuden alenemiseen.

PHOTOCATALYTIC OXIDATION OF AQUEOUS SOLUTIONS OF JET FUEL AND ICING INHIBITORS

The present paper is devoted to the results of experimental research undertaken into photocatalytical oxidation (PCO) of aqueous solutions of de-icing agents and aqueous extract of jet fuel. The report consists of introduction, literature review, description of materials and methods, discussion of results and conclusions. TiO2 was selected as a photocatalyst for the experiments with synthetic solutions of ethylene glycol, 2-ethoxyethanol and aqueous extract of jet fuel. To explain the PCO mechanisms affecting certain behaviour of de-icing agent under distinctive conditions, the following factors were studied: the impact of initial concentration of pollutant, the role of pH, the presence of tert-butanol as OH·-radicals scavenger and mineral admixtures. PCO under solar radiation performed in two ways: catalysed by irradiated TiO2 slurry or by TiO2 attached to buoyant hollow glass micro-spheres. Special attention was paid to the energy-saving PCO with reduced intensity mixing of the slurry. The effect of PCO was assessed by determination of residual chemical oxygen demand of solution (COD) and by measuring of concentration of glycols. The PCO process efficiency was assumed to be dependent on the TiO2 suspension fractional composition. Thus, the following effects of solutions’ media were viewed: presence of organic admixtures, pH influence, mixing mode during the PCO. The effects of mineral admixtures - Ca2+, Fe3+/2+, Mn2+, SO42- - that are often present in natural and wastewater systems or produced during the degradation of organic pollutants and which can affect the rate of PCO of de-icing agents, were also investigated.

Painatuksen UV-stabiilisuuden optimointi fenolihartsatussa paperissa

Tässä diplomityössä tutkittiin painetun paperin ja siitä fenoliformaldehydihartsilla impregnoimalla valmistetun pinnoituskalvon UV-stabiilisuuden parantamis-mahdollisuuksia. Työn kirjallisuusosassa käsitellään painetun pinnoituskalvon valmistusprosessia ja painatuksen UV-valonkestoon vaikuttavia tekijöitä. Painovärin pigmentti, sen määrä ja käsittely, painovärin sideaine sekä fenoliformaldehydihartsi ja sen lisäaineet vaikuttavat pinnoitetun betonoimisvanerin säänkesto-ominaisuuksiin. Erilaisilla epäorgaanisilla valkoisilla pigmenteillä ja kidemuodoilla on erilainen UV-valonkesto ja taitekerroin. Päällystämällä titaanidioksidi esimerkiksi alumiini- tai zirkoniumoksideilla sen UV-valonkestoa voidaan parantaa merkittävästi. UV-hajoaminen voidaan havaita painetun pinnoitteen liituuntumisena. Liituuntumista voidaan pitää veden ja hapen välisenä reaktiona, jota titaanidioksidi ja UV-säteily katalysoivat. Sen takia myös muiden valkoisten epäorgaanisten pigmenttien ominaisuuksia ja käyttöä selvitettiin. Kokeissa käytettiin yhdeksää eri painoväriä, kahta eri paksuista paperia ja kahta eri tyyppistä hartsia. Painovärejä ohennettiin vedellä ja paperin painopuolta vaihdeltiin. Kaikissa painatuksissa käytettiin kolmea eri rasterointiasteen laattaa, jolloin painovärin määrää paperissa saatiin vähennettyä. Painetuista papereista mitattiin densiteetti, värimäärä, pisara-absorptio vedellä ja kontaktikulma hartsilla. Myös painovärin tunkeumaa selvitettiin paperin poikkileikeistä tehtyjen SEM-kuvien avulla. Painetut paperit impregnoitiin fenoliformaldehydihartsilla kalvoksi. Pinnoituskalvot puristettiin vanerin pinnalle laboratoriopuristimella. Koekappaleet altistettiin UV-valolle, sateelle ja pakkaselle sääkaapissa 400 h ajan, mikä vastaa noin 1,5 vuotta ulkona Suomen oloissa. Kappaleista mitattiin kiilto, värinmuutos ja liituuntuminen. Pinnoitteen liituuntumista tapahtui vähiten niissä koepisteissä, joissa painatus oli tehty 30 % rasteroidulla laatallaSäänkestävä TiO2 osoittautui hyväksi, mutta myös ZnO-pigmentillä saatiin hyviä tuloksia. ZnO-koepisteessä liituuntumisreaktio ei ole niin voimakkaasti katalysoitu kuin TiO2-koepisteissä. Paksun paperin painatuspuolella näytti olevan merkitystä säänkestoon. Huopapuolelle painettuna pinnoitteen liituuntuminen oli vähäisempää

Gas-phase photocatalytic oxidation of volatile organic compounds

Substances emitted into the atmosphere by human activities in urban and industrial areas cause environmental problems such as air quality degradation, respiratory diseases, climate change, global warming, and stratospheric ozone depletion. Volatile organic compounds (VOCs) are major air pollutants, emitted largely by industry, transportation and households. Many VOCs are toxic, and some are considered to be carcinogenic, mutagenic, or teratogenic. A wide spectrum of VOCs is readily oxidized photocatalytically. Photocatalytic oxidation (PCO) over titanium dioxide may present a potential alternative to air treatment strategies currently in use, such as adsorption and thermal treatment, due to its advantageous activity under ambient conditions, although higher but still mild temperatures may also be applied. The objective of the present research was to disclose routes of chemical reactions, estimate the kinetics and the sensitivity of gas-phase PCO to reaction conditions in respect of air pollutants containing heteroatoms in their molecules. Deactivation of the photocatalyst and restoration of its activity was also taken under consideration to assess the practical possibility of the application of PCO to the treatment of air polluted with VOCs. UV-irradiated titanium dioxide was selected as a photocatalyst for its chemical inertness, non-toxic character and low cost. In the present work Degussa P25 TiO2 photocatalyst was mostly used. In transient studies platinized TiO2 was also studied. The experimental research into PCO of following VOCs was undertaken: - methyl tert-butyl ether (MTBE) as the basic oxygenated motor fuel additive and, thus, a major non-biodegradable pollutant of groundwater; - tert-butyl alcohol (TBA) as the primary product of MTBE hydrolysis and PCO; - ethyl mercaptan (ethanethiol) as one of the reduced sulphur pungent air pollutants in the pulp-and-paper industry; - methylamine (MA) and dimethylamine (DMA) as the amino compounds often emitted by various industries. The PCO of VOCs was studied using a continuous-flow mode. The PCO of MTBE and TBA was also studied by transient mode, in which carbon dioxide, water, and acetone were identified as the main gas-phase products. The volatile products of thermal catalytic oxidation (TCO) of MTBE included 2-methyl-1-propene (2-MP), carbon monoxide, carbon dioxide and water; TBA decomposed to 2-MP and water. Continuous PCO of 4 TBA proceeded faster in humid air than dry air. MTBE oxidation, however, was less sensitive to humidity. The TiO2 catalyst was stable during continuous PCO of MTBE and TBA above 373 K, but gradually lost activity below 373 K; the catalyst could be regenerated by UV irradiation in the absence of gas-phase VOCs. Sulphur dioxide, carbon monoxide, carbon dioxide and water were identified as ultimate products of PCO of ethanethiol. Acetic acid was identified as a photocatalytic oxidation by-product. The limits of ethanethiol concentration and temperature, at which the reactor performance was stable for indefinite time, were established. The apparent reaction kinetics appeared to be independent of the reaction temperature within the studied limits, 373 to 453 K. The catalyst was completely and irreversibly deactivated with ethanethiol TCO. Volatile PCO products of MA included ammonia, nitrogen dioxide, nitrous oxide, carbon dioxide and water. Formamide was observed among DMA PCO products together with others similar to the ones of MA. TCO for both substances resulted in the formation of ammonia, hydrogen cyanide, carbon monoxide, carbon dioxide and water. No deactivation of the photocatalyst during the multiple long-run experiments was observed at the concentrations and temperatures used in the study. PCO of MA was also studied in the aqueous phase. Maximum efficiency was achieved in an alkaline media, where MA exhibited high fugitivity. Two mechanisms of aqueous PCO – decomposition to formate and ammonia, and oxidation of organic nitrogen directly to nitrite - lead ultimately to carbon dioxide, water, ammonia and nitrate: formate and nitrite were observed as intermediates. A part of the ammonia formed in the reaction was oxidized to nitrite and nitrate. This finding helped in better understanding of the gasphase PCO pathways. The PCO kinetic data for VOCs fitted well to the monomolecular Langmuir- Hinshelwood (L-H) model, whereas TCO kinetic behaviour matched the first order process for volatile amines and the L-H model for others. It should be noted that both LH and the first order equations were only the data fit, not the real description of the reaction kinetics. The dependence of the kinetic constants on temperature was established in the form of an Arrhenius equation.

Photocatalytic Activity of Nanostructured Anatase Coatings Obtained by Cold Gas Spray

This article describes a photocatalytic nanostructured anatase coating deposited by cold gas spray (CGS)supported on titanium sub-oxide (TiO22x) coatings obtained by atmospheric plasma spray (APS) onto stainless steel cylinders. The photocatalytic coating was homogeneous and preserved the composition and nanostructure of the starting powder. The inner titanium sub-oxide coating favored the deposition of anatase particles in the solid state. Agglomerated nano-TiO2 particles fragmented when impacting onto the hard surface of the APS TiO22x bond coat. The rough surface provided by APS provided an ideal scenario for entrapping the nanostructured particles, which may be adhered onto the bond coat due to chemical bonding; a possible bonding mechanism is described. Photocatalytic experiments showed that CGS nano-TiO2 coating was active for photodegrading phenol and formic acid under aqueous conditions. The results were similar to the performance obtained by competitor technologies and materials such as dip-coating P25 photocatalysts. Disparity in the final performance of the photoactive materials may have been caused by differences in grain size and the crystalline composition of titanium dioxide.

Suitability of human and mammalian cells of different origin for the assessment of genotoxicity of metal and polymeric engineered nanoparticles.

Nanogenotoxicity is a crucial endpoint in safety testing of nanomaterials as it addresses potential mutagenicity, which has implications for risks of both genetic disease and carcinogenesis. Within the NanoTEST project, we investigated the genotoxic potential of well-characterised nanoparticles (NPs): titanium dioxide (TiO2) NPs of nominal size 20 nm, iron oxide (8 nm) both uncoated (U-Fe3O4) and oleic acid coated (OC-Fe3O4), rhodamine-labelled amorphous silica 25 (Fl-25 SiO2) and 50 nm (Fl-50 SiO) and polylactic glycolic acid polyethylene oxide polymeric NPs - as well as Endorem® as a negative control for detection of strand breaks and oxidised DNA lesions with the alkaline comet assay. Using primary cells and cell lines derived from blood (human lymphocytes and lymphoblastoid TK6 cells), vascular/central nervous system (human endothelial human cerebral endothelial cells), liver (rat hepatocytes and Kupffer cells), kidney (monkey Cos-1 and human HEK293 cells), lung (human bronchial 16HBE14o cells) and placenta (human BeWo b30), we were interested in which in vitro cell model is sufficient to detect positive (genotoxic) and negative (non-genotoxic) responses. All in vitro studies were harmonized, i.e. NPs from the same batch, and identical dispersion protocols (for TiO2 NPs, two dispersions were used), exposure time, concentration range, culture conditions and time-courses were used. The results from the statistical evaluation show that OC-Fe3O4 and TiO2 NPs are genotoxic in the experimental conditions used. When all NPs were included in the analysis, no differences were seen among cell lines - demonstrating the usefulness of the assay in all cells to identify genotoxic and non-genotoxic NPs. The TK6 cells, human lymphocytes, BeWo b30 and kidney cells seem to be the most reliable for detecting a dose-response.

A fotocatálise heterogênea e sua aplicação ambiental

This article gives some basic principles of heterogeneous photocatalysis using titanium dioxide as photocatalyst and the state of art of its applications to the abatement of aqueous and atmospheric pollutants.

Dióxido de titânio sol-gel: propriedades e comportamento eletrocrômico

Titanium dioxide was prepared by hydrolysis and polycondensation of titanium tetraisopropoxide. TiO2 films were obtained by spin coating of the precursor solution on ITO substractes (glass covered with indium doped tin oxide). Films were prepared using different temperatures and hydrochloric acid contents. The effect of the drying temperature of the films (100 or 400ºC) was also investigated. TiO2 films were characterized by cyclic voltammetry, chronoamperometry, ultraviolete-visible spectroscopy, scanning electron microscopy and X-ray diffractrometry.

Inativação por oxidação fotocatalítica de Escherichia coli e Pseudomonas sp.

This study presents the bactericidal activity of titanium dioxide photocatalysis, using as model Escherichia coli and Pseudomonas sp. cells. It was observed that the process efficiency is related to initial cell concentration, light intensity, UV irradiation exposure time, TiO2 concentration increase. The ultimate removal efficiency was above 99.9%.

Imobilização de TiO2 em concreto: fotodegradação de clorofórmio e fenol

TiO2 immobilization on concrete was studied using mixtures with cement, varnish and resin. The UV radiation sources were a germicide UV lamp and solar light. Aqueous solutions of chloroform (CHCl3) and of phenol were prepared and recirculated over the TiO2 immobilized surfaces. The immobilized TiO2 surfaces showed better photocatalytic efficiency for phenol degradation compared to the control. For CHCl3, the presence or absence of the catalyst did not cause any significant difference to its degradation efficiency. The micrographic results showed a more homogeneous surface for TiO2 immobilized in resin and varnish.

Efeitos dos parâmetros operacionais na fotodegradação do azo corante direct red 23 na interface dióxido de titânio/água

The decolorization and degradation of direct red 23 azo dye have been investigated in aqueous suspension of titanium dioxide under artificial irradiation. The effects of some operational parameters such as azo dye concentration, catalyst loading, and solution pH were investigated at 30.0 ºC and optimized values were obtained. The first-order kinetic model was used to discuss the results. The UV-Vis spectra changes showed that the azo dye sample, collected after 6 h irradiation, was 98% decolorized while the residual total carbon was 97.9% degraded, indicating simultaneous photodecolorization and degradation.

Obtenção de filmes finos de TiO2 nanoestruturado pelo método dos precursores poliméricos

This work focuses in optimizing setup for obtaining TiO2 thin films by polymeric precursor route due to its advantages on stoichiometric and morphological control. Precursor stoichiometry, synthesis pH, solids concentration and rotation speed at deposition were optimized evaluating thin films morphology and thickness. Thermogravimetry and RMN were applied for precursor's characterization and AFM, XRD and ellipsometry for thin films evaluation. Results showed successful attainment of homogeneous nanocrystalline anatase TiO2 thin films with outstanding control over morphological characteristics, mean grain size of 17 nm, packing densities between 57 and 75%, estimated surface areas of 90 m²/g and monolayers thickness within 20 and 128 nm.