Abstract  In order to understand how nanoparticles (NPs <100 nm) interact with cellular systems, potentially causing adverse effects, it is important to be able to detect and localize them within cells. Due to the small size of NPs, transmission electron microscopy (TEM) is an appropriate technique to use for visualizing NPs inside cells, since light microscopy fails to resolve them at a single particle level. However, the presence of other cellular and non-cellular nano-sized structures in TEM cell samples, which may resemble NPs in size, morphology and electron density, can obstruct the precise intracellular identification of NPs. Therefore, elemental analysis is recommended to confirm the presence of NPs inside the cell. The present study highlights the necessity to perform elemental analysis, specifically energy filtering TEM, to confirm intracellular NP localization using the example of quantum dots (QDs). Recently, QDs have gained increased attention due to their fluorescent characteristics, and possible applications for biomedical imaging have been suggested. Nevertheless, potential adverse effects cannot be excluded and some studies point to a correlation between intracellular particle localization and toxic effects. J774.A1 murine macrophage-like cells were exposed to NH2 polyethylene (PEG) QDs and elemental co-localization analysis of two elements present in the QDs (sulfur and cadmium) was performed on putative intracellular QDs with electron spectroscopic imaging (ESI). Both elements were shown on a single particle level and QDs were confirmed to be located inside intracellular vesicles. Nevertheless, ESI analysis showed that not all nano-sized structures, initially identified as QDs, were confirmed. This observation emphasizes the necessity to perform elemental analysis when investigating intracellular NP localization using TEM.

Abstract  This report consists of charts of toxic pollutants with the recommended water quality criteria listed for such things as freshwater, saltwater, human consumption, etc.

Abstract  This study aims at investigating feasibility and challenges associated with conducting a human health risk assessment for nano-silver based on the open literature by following an approach similar to a classical regulatory risk assessment. Gaps in the available data set, both in relation to exposures and hazard, do not allow reaching any definite conclusions that could be used for regulatory decision making. Results show that repeated inhalation in the workplace and possibly consumer inhalation may cause risks. Also (uncontrolled) nano-silver drug intake and burn treatment of large parts of the body with wound dressings may cause risks. Main future work should focus on generating occupational and consumer exposure data, as well as toxicity data on absorption (are particles or only ions absorbed?), information on genetoxicity, and further information on the toxicity following inhalation exposure to sizes and agglomeration states as uncounted in the workplace.

Abstract  Some adjustments are needed in the European chemicals legislation REACH to assess and control the risks of nanomaterials. The information on substances to be provided under REACH is not sufficient to determine the specific properties of nanomaterials, nor to assess how these properties affect their behaviour and effects in humans and the environment. RIVM concluded this following research into the suitability of REACH for nanomaterials. RIVM therefore proposes an adapted set of minimum information requirements, to be applied to all nanomaterials to be registered under REACH, independent of their volume of production and import. These requirements allow a risk assessment of nanomaterials. Over the last years the use of nanomaterials has strongly increased. As yet, nanomaterials are defined as substances of which the discrete parts have at least one dimension smaller than one hundred nanometres. Due to their nanosize they have specific properties. Legislation should focus on controlling the potential hazards and risks of these nanomaterials. By conducting a hypothetical registration of nanosilver it was investigated whether REACH is suitable for assessing the safe use of nanomaterials. From this it appeared that no definition of a nanomaterial is present, and that a relevant measure for expressing harmfulness and exposure is as yet not known. In addition, the standard information requirements are insufficient to assess hazard and exposure. They are also insufficient for a proper characterisation of the nanomaterial. Consequently, it cannot be determined to what extent the nanoform of a substance corresponds to the non-nanoform of the same substance. Furthermore, it is unclear whether current risk reduction measures and extrapolation methods in risk assessment, as established for non-nanomaterials, are applicable to nanomaterials.

Abstract  Exhaled nitric oxide (eNO) levels are increased in untreated or unstable asthma and measurements can be made easily. Our aim was to assess the usefulness of eNO for diagnosing and predicting loss of control (LOC) in asthma following steroid withdrawal. Comparisons were made against sputum eosinophils and airway hyperresponsiveness (AHR) to hypertonic saline (4.5%). Seventy-eight patients with mild/moderate asthma had their inhaled steroid therapy withdrawn until LOC occurred or for a maximum of 6 wk. Sixty (77.9%) developed LOC. There were highly significant correlations between the changes in eNO and symptoms (p < 0.0001), FEV(1) (p < 0.002), sputum eosinophils (p < 0.0002), and saline PD(15) (p < 0.0002), and there were significant differences between LOC and no LOC groups. Both single measurements and changes of eNO (10 ppb, 15 ppb, or an increase of > 60% over baseline) had positive predictive values that ranged from 80 to 90% for predicting and diagnosing LOC. These values were similar to those obtained using sputum eosinophils and saline PD(15) measurements. We conclude that eNO measurements are as useful as induced sputum analysis and AHR in assessing airway inflammation, with the advantage that they are easy to perform.

Abstract  Silver nanoparticles stabilized by a well-known antibacterial surfactant benzyldimethyl[3-(myristoylamino)propyl]ammonium chloride (Myramistin®) were produced for the first time by borohydride reduction of silver chloride sol in water. Stable aqueous dispersions of silver nanoparticles without evident precipitation for several months could be obtained. In vitro bactericidal tests showed that Myramistin® capped silver NPs exhibited notable activity against six different microorganisms—gram-positive and gram-negative bacteria, yeasts and fungi. The activity was up to 20 times higher (against E. coli) compared to Myramistin® at the same concentrations and on average 2 times higher if compared with citrate-stabilized NPs.

Abstract  Algal bioassays for heavy metals can detect low levels in the environment, for example, 0.01 ppm for silver. Algae respond to increasing levels of heavy metals such as copper, nickel, mercury, silver, or cadmium by reduction of growth rate. Occasionally, the response to nontoxic metals is an increase in growth rate. At very low concentrations some potentially toxic metals may be necessary micronutrients. Algal species differ quite markedly in their sensitivity to heavy metals. Combined effects of two or more metals at toxic concentrations may be synergistic (for example, copper-nickel) or antagonistic (for example, cadmium-selenium). The critical concentrations for toxicity of a particular metal may be different at different times during the growth of an algal culture, as well as being dependent upon other chemical and physical conditions. Algal cells appear to markedly concentrate metals from solution, even at concentrations of these metals in the medium which do not apparently inhibit cell division. Bioassays provide the only direct method for assessing the biological availability of metals in solution. Algae isolated from metal-polluted lakes appear to have evolved specific metal tolerances. These "tolerant" algae actually accumulate more of the metals concerned than do their "nontolerant" relatives. Correlations between fish toxicity tests and algal bioassays may allow the relatively expensive fish testing schemes to be replaced by simple and cheaper algal bioassays.

Abstract  The report was a result of an effort by experts from government, industry and the public interest community to examine the path of a number of hypothetical nanotechnology food packaging applications through the current regulatory system. The regulatory system for food packaging is scientifically rigorous and extraordinarily complex, both legally and scientifically. This first-of-its-kind analysis provides a better understanding of the potential regulatory issues on the horizon for nanotechnology-enabled packaging – an advantage for industry, consumers and regulatory agencies such as FDA and the Environmental Protection Agency (EPA).

Abstract  Exposure assessment is crucial for risk assessment for nanomaterials. We propose a framework to aid exposure assessment in consumer products. We determined the location of the nanomaterials and the chemical identify of the 580 products listed in the inventory maintained by the Woodrow Wilson International Center for Scholars, of which 37% used nanoparticles suspended in liquids, whereas <1% contained "free airborne nanoparticles". C(60) is currently only used as suspended nanoparticles in liquids and nanosilver is used more as surface bound nanoparticles than as particles suspended in liquids. Based on the location of the nanostructure we were able to further group the products into categories of: (1) expected, (2) possible, and (3) no expected exposure. Most products fall into the category of expected exposure, but we were not able to complete a quantitative exposure assessment mainly due to the lack of information on the concentration of the nanomaterial in the products--a problem that regulators and industry will have to address if we are to have realistic exposure assessment in the future. To illustrate the workability of our procedure, we applied it to four product scenarios using the best estimates available and/or worst-case assumptions. Using the best estimates available and/or worst-case assumptions we estimated the consumer exposure to be 26, 15, and 44 microg kg(-1) bw year(-1) for a facial lotion, a fluid product, and a spray product containing nanoparticles, respectively. The application of sun lotion containing 2% nanoparticles result in an exposure of 56.7 mg kg(-1) bw d(-1) for a 2-year-old child, if the amounts applied correspond to the European Commission recommendations on use of sunscreen.

Abstract  There is an emerging literature reporting toxic effects of manufactured nanomaterials (NMs) and nanoparticles (NPs) in fish, but the mechanistic basis of both exposure and effect are poorly understood. This paper critically evaluates some of the founding assumptions in fish toxicology, and likely mechanisms of absorption, distribution, metabolism and excretion (ADME) of NPs in fish compared to other chemicals. Then, using a case study approach, the paper compares these assumptions for two different NPs; TiO2 and C60 fullerenes. Adsorption of NPs onto the gill surface will involve similar processes in the gill microenvironment and mucus layer to other substances, but the uptake mechanisms for NPs by epithelial cells are more likely to occur via vesicular processes (e.g., endocytosis) than uptake on membrane transporters or by diffusion through the cell membranes. Target organs may include the gills, gut, liver and sometimes the brain. Information on metabolism and excretion of NPs in fish is limited; but hepatic excretion into the bile seems a more likely mechanism, rather than mainly by renal or branchial excretion. TiO2 and C60 share some common chemical properties that appear to be associated with some similar toxic effects, but there are also differences, that highlight the notion that chemical reactivity can inform toxic effect of NPs in a fundamentally similar way to other chemicals. In this paper we identify many knowledge gaps including the lack of field observations on fish and other wildlife species for exposure and effects of manufactured NMs. Systematic studies of the abiotic factors that influence bioavailability, and investigation of the cell biology that informs on the mechanisms of metabolism and excretion of NMs, will greatly advance our understanding of the potential for adverse effects. There are also opportunities to apply existing tools and techniques to fundamental studies of fish toxicology with NPs, such as perfused organs and fish cell culture systems.

Abstract  Environmental toxicologists, chemists and social scientists have identified three priorities for research into the impact of engineered nanoparticles on the environment.

Abstract  It is inevitable that, during their use, engineered nanoparticles will be released into soils and waters. There is therefore increasing concern over the potential impacts of engineered nanoparticles in the environment on aquatic and terrestrial organisms and on human health. Once released into the environment, engineered nanoparticles will aggregate to some degree; they might also associate with suspended solids, sediment, be accumulated by organisms and enter drinking water sources and food materials. These fate processes are dependent on the characteristics of the particle and the characteristics of the environmental system. A range of ecotoxicological effects have also been reported, including effects on microbes, plants, invertebrates and fish. Although available data indicate that current risks of engineered nanoparticles in the environment to environmental and human health are probably low, our knowledge of the potential impacts of engineered nanoparticles in the environment on human health is still limited. There is therefore a need for continued work to develop an understanding of the exposure levels for engineered nanoparticles in environmental systems and to begin to explore the implications of these levels in terms of the ecosystem and human health. This will require research in a range of areas, including detection and characterization, environmental fate and transport, ecotoxicology and toxicology.

Abstract  We investigated the mechanism by which transferrin-coated gold nanoparticles (Au NP) of different sizes and shapes entered mammalian cells. We determined that transferrin-coated Au NP entered the cells via clathrin-mediated endocytosis pathway. The NPs exocytosed out of the cells in a linear relationship to size. This was different than the relationship between uptake and size. Furthermore, we developed a mathematical equation to predict the relationship of size versus exocytosis for different cell lines. These studies will provide guidelines for developing NPs for imaging and drug delivery applications, which will require "controlling" NP accumulation rate. These studies will also have implications in determining nanotoxicity.

Abstract  Ultrafine particles (UFP, particles <100 nm) are ubiquitous in ambient urban and indoor air from multiple sources and may contribute to adverse respiratory and cardiovascular effects of particulate matter (PM). Depending on their particle size, inhaled UFP are efficiently deposited in nasal, tracheobronchial, and alveolar regions due to diffusion. Our previous rat studies have shown that UFP can translocate to interstitial sites in the respiratory tract as well as to extrapulmonary organs such as liver within 4 to 24h postexposure. There were also indications that the olfactory bulb of the brain was targeted. Our objective in this follow-up study, therefore, was to determine whether translocation of inhaled ultrafine solid particles to regions of the brain takes place, hypothesizing that UFP depositing on the olfactory mucosa of the nasal region will translocate along the olfactory nerve into the olfactory bulb. This should result in significant increases in that region on the days following the exposure as opposed to other areas of the central nervous system (CNS). We generated ultrafine elemental 13C particles (CMD = 36 nm; GSD = 1.66) from [13C] graphite rods by electric spark discharge in an argon atmosphere at a concentration of 160 microg/m3. Rats were exposed for 6 h, and lungs, cerebrum, cerebellum and olfactory bulbs were removed 1, 3, 5, and 7 days after exposure. 13C concentrations were determined by isotope ratio mass spectroscopy and compared to background 13C levels of sham-exposed controls (day 0). The background corrected pulmonary 13C added as ultrafine 13C particles on day 1 postexposure was 1.34 microg/lung. Lung 13C concentration decreased from 1.39 microg/g (day 1) to 0.59 microg/g by 7 days postexposure. There was a significant and persistent increase in added 13C in the olfactory bulb of 0.35 microg/g on day 1, which increased to 0.43 microg/g by day 7. Day 1 13C concentrations of cerebrum and cerebellum were also significantly increased but the increase was inconsistent, significant only on one additional day of the postexposure period, possibly reflecting translocation across the blood-brain barrier in certain brain regions. The increases in olfactory bulbs are consistent with earlier studies in nonhuman primates and rodents that demonstrated that intranasally instilled solid UFP translocate along axons of the olfactory nerve into the CNS. We conclude from our study that the CNS can be targeted by airborne solid ultrafine particles and that the most likely mechanism is from deposits on the olfactory mucosa of the nasopharyngeal region of the respiratory tract and subsequent translocation via the olfactory nerve. Depending on particle size, >50% of inhaled UFP can be depositing in the nasopharyngeal region during nasal breathing. Preliminary estimates from the present results show that ~20% of the UFP deposited on the olfactory mucosa of the rat can be translocated to the olfactory bulb. Such neuronal translocation constitutes an additional not generally recognized clearance pathway for inhaled solid UFP, whose significance for humans, however, still needs to be established. It could provide a portal of entry into the CNS for solid UFP, circumventing the tight blood-brain barrier. Whether this translocation of inhaled UFP can cause CNS effects needs to be determined in future studies.

Abstract  The deterioration of indoor air quality because of airborne bacterial consortia is a widespread environmental problem. The main objective of this study is to evaluate the feasibility of applying nanoscale silver-doped titanium dioxide (TiO2) photocatalysis (nano-Ag/TiO2) particles as a photocatalyst to enhance the disinfecting capability of bacterial restraining equipment in a medical nursing institution. A full-scale, on-site study was initiated to test the effectiveness of adding the photocatalyst to various air quality control equipments; that is, fixed full-covering type (FF type), underneath swinging type (US type), and lateral swinging type (LS type). Results from this study indicate that the FF type has the best bacterial restraining rate (81%) as compared with the LS type (66%) and the US type (61%). Similar results are obtained at different heights (90 and 180 cm) of the nursing institutions, revealing that freshly disinfected air is provided regardless of location as long as the air quality control equipment is in operation. The proposed equipment can disinfect air to effectively restrain bacteria as demonstrated on-site in medical nursing institutions; the results will also be for designing integrated, commercialized, large-scale applications in the future.

Abstract  Nano-silver (Ag) with antimicrobial activity is by far the most commercialized nano-compound. The hazards associated with human exposure to nanosized-silver should be investigated to facilitate the risk assessment process. Recent studies have shown that inflammatory, oxidative, genotoxic, and cytotoxic consequences are associated with silver particulate exposure, and are inherently linked. In the present study, the cytotoxicity and genotoxicity of nano-silver were investigated using the dye exclusion assay, the comet assay, and the mouse lymphoma thymidine kinase (tk+/−) gene mutation assay (MLA). IC20 values of nano-silver in L5178Y cells were determined the concentration of 3,769.53 μg/mL and 1,796.88 μg/mL with and without S-9, respectively. And in BEAS-2B cell, IC20 values were calculated to 1,171.88 μg/mL and 761.72 μg/mL with and without S-9, respectively. From these results, nano-silver was more cytotoxic in absence of S-9 metabolic activation system and at the BEAS-2B cells. In the comet assay, L5178Y cells and BEAS-2B cells were treated with nano-silver which significantly increased >2-fold tail moment with and without S-9. However, the mutant frequencies in the nano-silver treated L5178Y cells were slightly increased but not significant compared to the vehicle controls with and without S-9. The results of this study indicate that nano-silver can cause primary DNA damage and cytotoxicity but not mutagenicity in cultured mammalian cells.

Abstract  Nanocomposite thin films (170 nm), composed of silver nanoparticles enclosed in an organosilicon matrix, were deposited onto stainless steel, with the aim of preventing biofilm formation. The film deposition was carried out under cold plasma conditions, combining radiofrequency (RF) glow discharge fed with argon and hexamethyldisiloxane and simultaneous silver sputtering. XPS and ToF-SIMS were used to characterize Ag-organosilicon films in native form and after ageing in saline solution (NaCl 0.15 M), in order to further correlate their lifetime with their anti-fouling properties. Two coatings with significantly different silver contents (7.5% and 20.3%) were tested. Surface analysis confirmed the presence of metallic silver in the pristine coating and revealed significant modifications after immersion in the saline solution. Two different ageing mechanisms were observed, depending on the initial silver concentration in the film. For the sample exhibiting the low silver content (7.5%), the metal amount decreased at the surface in contact with the solution, due to the release of silver from the coating. As a result, after a 2-day exposure, silver nanoparticles located at the extreme surface were entirely released, whereas silver is still present in the inner part of the film. The coating thickness was not modified during ageing. In contrast, for the high silver content film (20.3%), the thickness decreased with immersion time, due to significant silver release and matrix erosion, assigned to a percolation-like effect. However, after 18 days of immersion, the delamination process stopped and a thin strongly bounded layer remained on the stainless steel surface.

Abstract  Silver nanoparticles (AgNPs) have emerged as an important class of nanomaterials and are currently used in a wide range of industrial and commercial applications. This has caused increasing concern about their effects on the environment and to human health. Using Japanese medaka (Oryzias latipes) at early-life stages as experimental models, the developmental toxicity of silver nanoparticles was investigated following exposure to 100–1000 μg/L homogeneously dispersed AgNPs for 70 days, and developmental endpoints were evaluated by microscopy during embryonic, larval and juvenile stages of development in medaka. Meanwhile, histopathological changes in the larval eye were evaluated. Retarded development and reduced pigmentation were observed in the treated embryos by AgNPs at high concentrations (≥400 μg/L). Maximum width of the optic tectum, as an indicator of midbrain development, decreased significantly in a dose-related manner. Furthermore, silver nanoparticles exposure at all concentrations induced a variety of morphological malformations such as edema, spinal abnormalities, finfold abnormalities, heart malformations and eye defects. Histopathological observations also confirmed the occurrence of abnormal eye development induced by AgNPs. The data showed non-linear or U-shaped dose–response patterns for growth retardation at 5 days of postfertilization, as well as the incidence of abnormalities. Preliminary results suggested that the developmental process of medaka may be affected by exposure to silver nanoparticles. Morphological abnormalities in early-life stages of medaka showed the potential developmental toxicities of silver nanoparticles. Further research should be focused on the mechanisms of developmental toxicity in fish exposed to silver nanoparticles.

Abstract  Nanoparticles with cores composed of inorganic materials such as noble, magnetic metals, their alloys and oxides, and semiconductors have been most studied and have vast potential for application in many different areas of biomedicine, from diagnostics to treatment of diseases. The effects of nanoparticles must be predictable and controllable, and deliver the desired result with minimum cytotoxicity. These criteria can be met by careful tailoring of the ligand shell, allowing stabilisation, specific targeting and recognition of biochemical species. For these reasons, this review is focused on the synthesis and biofunctionalisation of inorganic metal, semiconductor and magnetic nanoparticles for biomedical applications.

Abstract  Nanotoxicology is still a new discipline. In this Perspective, both its origins and its future trends are discussed. In particular, we note several issues we consider important for publications in this field.

Abstract  In this article, we represent a versatile and effective technique which using non-toxic chemicals to prepare stable aqueous dispersions of silver nanoparticles (NPs) via modified Tollens process. It was shown that as-prepared silver colloids consisted of finely-dispersed NPs with average diameter about 10 nm and a relatively narrow size distribution. Moreover, they could be stored very stable after several months without observation of aggregates or sedimentation. In comparison with previous works where Tollens process was being used, we for the first time applied UV-irradiation simultaneously with glucose reduction of silver salt through NPs preparation. The colloidal solutions of silver NPs were found to exhibit a high antibacterial activity against gram-negative Escherichia coli. The concentration of silver leading to a complete inhibition of bacteria growth was revealed as low as at 1.0 μg ml−1 and found much lower compared to earlier reports. These advantages of aqueous dispersions of silver NPs make them ideal for green industrial, medicinal, microbiological and other applications.

Abstract  The increasing use of manufactured nanoparticles ensures these materials will make their way into the environment. Silver nanoparticles in particular, due to use in a wide range of applications, have the potential to get into water systems, e.g., drinking water systems, ground water systems, estuaries, and/or lakes. One important question is what is the chemical and physical state of these nanoparticles in water? Are they present as isolated particles, agglomerates or dissolved ions, as this will dictate their fate and transport. Furthermore, does the chemical and physical state of the nanoparticles change as a function of size or differ from micron-sized particles of similar composition? In this study, an electrospray atomizer coupled to a scanning mobility particle sizer (ES-SMPS) is used to investigate the state of silver nanoparticles in water and aqueous nitric acid environments. Over the range of pH values investigated, 0.5–6.5, silver nanoparticles with a bimodal primary particle size distribution with the most intense peak at 5.0 ± 7.4 nm, as determined from transmission electron microscopy (TEM), show distinct size distributions indicating agglomeration between pH 6.5 and 3 and isolated nanoparticles at pH values from 2.5 to 1. At the lowest pH investigated, pH 0.5, there are no peaks detected by the SMPS, indicating complete nanoparticle dissolution. Further analysis of the solution shows dissolved Ag ions at a pH of 0.5. Interestingly, silver nanoparticle dissolution shows size dependent behavior as larger, micron-sized silver particles show no dissolution at this pH. Environmental implications of these results are discussed.