Cell membrane penetration and mitochondrial targeting by platinum-decorated ceria nanoparticles.

In this work we investigate the interaction between endothelial cells and nanoparticles emitted by catalytic converters. Although catalyst-derived particles are recognized as growing burden added to environmental pollution, very little is known about their health impact. We use platinum-decorated ceria nanoparticles as model compounds for the actual emitted particles and focus on their fast uptake and association with mitochondria, the cell's powerhouse. Using live-cell imaging and electron microscopy we clearly show that 46 nm platinum-decorated ceria nanoparticles can rapidly penetrate cell membranes and reach the cytosol. Moreover, if suitably targeted, these particles are able to selectively attach to mitochondria. These results are complemented by cytotoxicity assays, thus providing insights into the biological effects of these particles on cells. Interestingly, no permanent membrane disruption or any other significant adverse effects on cells were observed. The unusual uptake behavior observed for 46 nm nanoparticles was not observed for equivalent but larger 143 nm and 285 nm platinum-decorated particles. Our results demonstrate a remarkable particle size effect in which particles smaller than ∼50-100 nm escape the usual endocytic pathway and translocate directly into the cytosol, while particles larger than ∼150 nm are internalized by conventional endocytosis. Since the small particles are able to bypass endocytosis they could be explored as drug and gene delivery vehicles. Platinum-decorated nanoparticles are therefore highly interesting in the fields of nanotoxicology and nanomedicine.

Fate of cerium dioxide nanoparticles in endothelial cells: exocytosis.

Although cytotoxicity and endocytosis of nanoparticles have been the subject of numerous studies, investigations regarding exocytosis as an important mechanism to reduce intracellular nanoparticle accumulation are rather rare and there is a distinct lack of knowledge. The current study investigated the behavior of human microvascular endothelial cells to exocytose cerium dioxide (CeO2) nanoparticles (18.8 nm) by utilization of specific inhibitors [brefeldin A; nocodazole; methyl-ß-cyclodextrin (MßcD)] and different analytical methods (flow cytometry, transmission electron microscopy, inductively coupled plasma mass spectrometry). Overall, it was found that endothelial cells were able to release CeO2 nanoparticles via exocytosis after the migration of nanoparticle containing endosomes toward the plasma membrane. The exocytosis process occurred mainly by fusion of vesicular membranes with plasma membrane resulting in the discharge of vesicular content to extracellular environment. Nevertheless, it seems to be likely that nanoparticles present in the cytosol could leave the cells in a direct manner. MßcD treatment led to the strongest inhibition of the nanoparticle exocytosis indicating a significant role of the plasma membrane cholesterol content in the exocytosis process. Brefeldin A (inhibitor of Golgi-to-cell-surface-transport) caused a higher inhibitory effect on exocytosis than nocodazole (inhibitor of microtubules). Thus, the transfer from distal Golgi compartments to the cell surface influenced the exocytosis process of the CeO2 nanoparticles more than the microtubule-associated transport. In conclusion, endothelial cells, which came in contact with nanoparticles, e.g., after intravenously applied nano-based drugs, can regulate their intracellular nanoparticle amount, which is necessary to avoid adverse nanoparticle effects on cells.

Valence properties of Cu and Ru in titanium-substituted LnCu3Ru4O12+δ (Ln = La, Pr, Nd) investigated by XANES and TGA.

In the solid-solution series La(y)Cu(3)RuxTi(4-x)O(12+δ) (0 ≤x≤ 4) the Cu and Ru electronic states are highly correlated. With increasing Ru content x the system properties change from a paramagnetic insulator with colossal dielectric constant to a heavy-fermion metal. To further elucidate the occurring phase transitions, the valences of Cu and Ru have been investigated utilizing XANES measurements at the Cu-K and the Ru-K absorption edges. It was found that the Ru oxidation number is close to +4 in all samples, while the Cu valence linearly decreases from +2 for the titanate (x = 0) to +1.6 for the ruthenate (x = 4). Additional thermogravimetric measurements have been used to determine the oxygen content and rather high oxygen excesses up to δ≈ 0.7 (for x = 0.5) were obtained. The additional oxygen for x < 2 is required to compensate the constant Ru +4 valence. Our findings are in accordance with the reported phase transitions of the magnetic and transport properties. Both the valence shift and the shapes of the absorption edges suggest a change from localized to itinerant character of the Cu electronic states with increasing x, while the Ru electrons remain localized. Analogous results concerning the valences were found for the Pr(y)Cu(3)RuxTi(4-x)O(12+δ) and Nd(y)Cu(3)RuxTi(4-x)O(12+δ) solid-solution series.

Rapid degradation of zinc oxide nanoparticles by phosphate ions.

Zinc oxide nanoparticles are highly sensitive towards phosphate ions even at pH 7. Buffer solutions and cell culture media containing phosphate ions are able to destroy ZnO nanoparticles within a time span from less than one hour to one day. The driving force of the reaction is the formation of zinc phosphate of very low solubility. The morphology of the zinc oxide particles has only a minor influence on the kinetics of this reaction. Surface properties related to different production methods and the presence and absence of labelling with a perylene fluorescent dye are more important. Particles prepared under acidic conditions are more resistant than those obtained in basic or neutral reaction medium. Surprisingly, the presence of a SiO2 coating does not impede the degradation of the ZnO core. In contrast to phosphate ions, ß-glycerophosphate does not damage the ZnO nanoparticles. These findings should be taken into account when assessing the biological effects or the toxicology of zinc oxide nanoparticles.

Effects of the physicochemical properties of titanium dioxide nanoparticles, commonly used as sun protection agents, on microvascular endothelial cells.

Until now, the potential effects of titanium dioxide (TiO2) nanoparticles on endothelial cells are not well understood, despite their already wide usage. Therefore, the present work characterizes six TiO2 nanoparticle samples in the size range of 19 × 17 to 87 × 13 nm, which are commonly present in sun protection agents with respect to their physicochemical properties (size, shape, ζ-potential, agglomeration, sedimentation, surface coating, and surface area), their interactions with serum proteins and biological impact on human microvascular endothelial cells (relative cellular dehydrogenase activity, adenosine triphosphate content, and monocyte chemoattractant protein-1 release). We observed no association of nanoparticle morphology with the agglomeration and sedimentation behavior and no variations of the ζ-potential (-14 to -19 mV) in dependence on the surface coating. In general, the impact on endothelial cells was low and only detectable at concentrations of 100 µg/ml. Particles containing a rutile core and having rod-like shape had a stronger effect on cell metabolism than those with anatase core and elliptical shape (relative cellular dehydrogenase activity after 72 h: 60 vs. 90 %). Besides the morphology, the nanoparticle shell constitution was found to influence the metabolic activity of the cells. Upon cellular uptake, the nanoparticles were localized perinuclearly. Considering that in the in vivo situation endothelial cells would come in contact with considerably lower nanoparticle amounts than the lowest-observable adverse effects level (100 µg/ml), TiO2 nanoparticles can be considered as rather harmless to humans under the investigated conditions.

Synthesis and characterization of fluorescence-labelled silica core-shell and noble metal-decorated ceria nanoparticles.

The present review article covers work done in the cluster NPBIOMEM in the DFG priority programme SPP 1313 and focuses on synthesis and characterization of fluorescent silica and ceria nanoparticles. Synthetic methods for labelling of silica and polyorganosiloxane/silica core-shell nanoparticles with perylenediimide derivatives are described, as well as the modification of the shell with thiol groups. Photometric methods for the determination of the number of thiol groups and an estimate for the number of fluorescent molecules per nanoparticles, including a scattering correction, have been developed. Ceria nanoparticles decorated with noble metals (Pt, Pd, Rh) are models for the decomposition products of automobile catalytic converters which appear in the exhaust gases and finally interact with biological systems including humans. The control of the degree of agglomeration of small ceria nanoparticles is the basis for their synthesis. Almost monodisperse agglomerates (40 ± 4-260 ± 40 nm diameter) can be prepared and decorated with noble metal nanoparticles (2-5 nm diameter). Fluorescence labelling with ATTO 647N gave the model particles which are now under biophysical investigation.

Intake of silica nanoparticles by giant lipid vesicles: influence of particle size and thermodynamic membrane state.

The uptake of nanoparticles into cells often involves their engulfment by the plasma membrane and a fission of the latter. Understanding the physical mechanisms underlying these uptake processes may be achieved by the investigation of simple model systems that can be compared to theoretical models. Here, we present experiments on a massive uptake of silica nanoparticles by giant unilamellar lipid vesicles (GUVs). We find that this uptake process depends on the size of the particles as well as on the thermodynamic state of the lipid membrane. Our findings are discussed in the light of several theoretical models and indicate that these models have to be extended in order to capture the interaction between nanomaterials and biological membranes correctly.

A fast analysis method to quantify nanoparticle uptake on a single cell level.

AIM: This study examines the absolute quantification of particle uptake into cells. METHODS: We developed a novel method to analyze stacks of confocal fluorescence images of single cells interacting with nano-and micro-particles. Particle_in_Cell-3D is a freely available ImageJ macro. During the image analysis routine, single cells are reconstructed in 3D and split into two volumes - intracellular and the membrane region. Next, particles are localized and color-coded accordingly. The mean intensity of single particles, measured in calibration experiments, is used to determine the absolute number of particles. RESULTS: Particle_in_Cell-3D was successfully applied to measure the uptake of 80-nm mesoporous silica nanoparticles into HeLa cells. Furthermore, it was used to quantify the absolute number of 100-nm polystyrene nanoparticles forming agglomerates of up to five particles; the accuracy of these results was confirmed by super-resolution, stimulated emission depletion microscopy. CONCLUSION: Particle_in_Cell-3D is a fast and accurate method that allows the quantification of particle uptake into cells.

Selection of key ambient particulate variables for epidemiological studies - applying cluster and heatmap analyses as tools for data reduction.

The success of epidemiological studies depends on the use of appropriate exposure variables. The purpose of this study is to extract a relatively small selection of variables characterizing ambient particulate matter from a large measurement data set. The original data set comprised a total of 96 particulate matter variables that have been continuously measured since 2004 at an urban background aerosol monitoring site in the city of Augsburg, Germany. Many of the original variables were derived from measured particle size distribution (PSD) across the particle diameter range 3 nm to 10 µm, including size-segregated particle number concentration, particle length concentration, particle surface concentration and particle mass concentration. The data set was complemented by integral aerosol variables. These variables were measured by independent instruments, including black carbon, sulfate, particle active surface concentration and particle length concentration. It is obvious that such a large number of measured variables cannot be used in health effect analyses simultaneously. The aim of this study is a pre-screening and a selection of the key variables that will be used as input in forthcoming epidemiological studies. In this study, we present two methods of parameter selection and apply them to data from a two-year period from 2007 to 2008. We used the agglomerative hierarchical cluster method to find groups of similar variables. In total, we selected 15 key variables from 9 clusters which are recommended for epidemiological analyses. We also applied a two-dimensional visualization technique called "heatmap" analysis to the Spearman correlation matrix. 12 key variables were selected using this method. Moreover, the positive matrix factorization (PMF) method was applied to the PSD data to characterize the possible particle sources. Correlations between the variables and PMF factors were used to interpret the meaning of the cluster and the heatmap analyses.

Perylene-labeled silica nanoparticles: synthesis and characterization of three novel silica nanoparticle species for live-cell imaging.

The increasing exposure of humans to nanoscaled particles requires well-defined systems that enable the investigation of the toxicity of nanoparticles on the cellular level. To facilitate this, surface-labeled silica nanoparticles, nanoparticles with a labeled core and a silica shell, and a labeled nanoparticle network-all designed for live-cell imaging-are synthesized. The nanoparticles are functionalized with perylene derivatives. For this purpose, two different perylene species containing one or two reactive silica functionalities are prepared. The nanoparticles are studied by transmission electron microscopy, widefield and confocal fluorescence microscopy, as well as by fluorescence spectroscopy in combination with fluorescence anisotropy, in order to characterize the size and morphology of the nanoparticles and to prove the success and homogeneity of the labeling. Using spinning-disc confocal measurements, silica nanoparticles are demonstrated to be taken up by HeLa cells, and they are clearly detectable inside the cytoplasm of the cells.

On the presence of Fe(IV) in Fe-ZSM-5 and FeSrO3-x --unequivocal detection of the 3d4 spin system by resonant inelastic X-ray scattering.

The contribution of a 3d(4) spin configuration to the valence electronic structure of Fe compounds can be probed via spin-selective Fe K-pre-edge absorption spectra, using resonant inelastic X-ray scattering (RIXS). The 3d(4) configuration of Fe(IV) can be unequivocally detected even in a mixture with the high-spin 3d(5) configuration of Fe(III). This is demonstrated on the perovskite FeSrO(3-x) with formal oxidation state Fe(IV). When the technique was applied to an Fe-ZSM-5 catalyst during reaction with N(2)O, no 3d(4) configuration was detected. The formation of Fe(IV) upon reaction of Fe-ZSM-5 with N(2)O can, therefore, be ruled out.

Structural and magnetic properties of hexagonal perovskites La1.2Sr2.7MO7.33 (M = Ru, Ir) containing peroxide ions.

The structures of the new compound La(1.2)Sr(2.7)IrO(7.33) and the recently discovered La(1.2)Sr(2.7)RuO(7.33) have been solved using a combination of X-ray and neutron diffraction. Both compounds crystallize in the trigonal space group Rm and consist of isolated MO6 (M = Ru, Ir) octahedra, which are arranged in well-defined hexagonal perovskite slabs. These slabs are separated by (Sr2O(1+delta)) layers containing both O2- and (O2)2- ions. The composition can therefore be written as La(1.2)Sr(2.7)MO(7-delta)(O2)delta with delta = 0.33. Results of the magnetic susceptibility and XANES measurements show that the transition metal cations are in a pentavalent state. While in La(1.2)Sr(2.7)RuO(7.33) an antiferromagnetic interaction between the Ru5+ ions is found, La(1.2)Sr(2.7)IrO(7.33) shows a very small temperature-independent paramagnetism down to 1.8 K due to the strong spin-orbit coupling characteristic for the 5d element iridium.

Powder neutron diffraction of SrNbO2N at room temperature and 1.5 K.

The structure of strontium niobium dioxygen nitride, SrNbO2N, has been solved by powder neutron diffraction at room temperature and 1.5 K. SrNbO2N crystallizes in the tetragonal space group I4/mcm, with a = 5.7056 (4) and c = 8.1002 (9) A at room temperature, and a = 5.6938 (4) and c = 8.0974 (8) A at 1.5 K. The crystal structure is derived from the cubic perovskite archetype by a slight rotation of the Nb(O,N)6 octahedra with respect to the tetragonal axis. A partially ordered distribution of oxygen and nitrogen on the anionic sites was found.

Nanotubes in Si-doped titanium dioxide.

Silicon-doped metal oxide nanotubes are formed in Si-doped titanium dioxide prepared by a sol-gel technique.