Validation of weak biological effects by round robin experiments: cytotoxicity/biocompatibility of SiO2 and polymer nanoparticles in HepG2 cells.

All over the world, different types of nanomaterials with a diversified spectrum of applications are designed and developed, especially in the field of nanomedicine. The great variety of nanoparticles (NPs), in vitro test systems and cell lines led to a vast amount of publications with conflicting data. To identify the decisive principles of these variabilities, we conducted an intercomparison study of collaborating laboratories within the German DFG Priority Program SPP1313, using well-defined experimental parameters and well-characterized NPs. The participants analyzed the in vitro biocompatibility of silica and polymer NPs on human hepatoma HepG2 cells. Nanoparticle mediated effects on cell metabolism, internalization, and inflammation were measured. All laboratories showed that both nanoparticle formulations were internalized and had a low cytotoxicity profile. Interestingly, small variations in nanoparticle preparation, cell handling and the type of culture slide influenced the nanoparticle stability and the outcomes of cell assays. The round robin test demonstrated the importance of the use of clearly defined and characterized NPs and parameters for reproducible results across laboratories. Comparative analyses of in vitro screening methods performed in multiple laboratories are absolutely essential to establish robust standard operation procedure as a prerequisite for sound hazard assessment of nanomaterials.

PVP-coated, negatively charged silver nanoparticles: A multi-center study of their physicochemical characteristics, cell culture and in vivo experiments.

PVP-capped silver nanoparticles with a diameter of the metallic core of 70 nm, a hydrodynamic diameter of 120 nm and a zeta potential of -20 mV were prepared and investigated with regard to their biological activity. This review summarizes the physicochemical properties (dissolution, protein adsorption, dispersability) of these nanoparticles and the cellular consequences of the exposure of a broad range of biological test systems to this defined type of silver nanoparticles. Silver nanoparticles dissolve in water in the presence of oxygen. In addition, in biological media (i.e., in the presence of proteins) the surface of silver nanoparticles is rapidly coated by a protein corona that influences their physicochemical and biological properties including cellular uptake. Silver nanoparticles are taken up by cell-type specific endocytosis pathways as demonstrated for hMSC, primary T-cells, primary monocytes, and astrocytes. A visualization of particles inside cells is possible by X-ray microscopy, fluorescence microscopy, and combined FIB/SEM analysis. By staining organelles, their localization inside the cell can be additionally determined. While primary brain astrocytes are shown to be fairly tolerant toward silver nanoparticles, silver nanoparticles induce the formation of DNA double-strand-breaks (DSB) and lead to chromosomal aberrations and sister-chromatid exchanges in Chinese hamster fibroblast cell lines (CHO9, K1, V79B). An exposure of rats to silver nanoparticles in vivo induced a moderate pulmonary toxicity, however, only at rather high concentrations. The same was found in precision-cut lung slices of rats in which silver nanoparticles remained mainly at the tissue surface. In a human 3D triple-cell culture model consisting of three cell types (alveolar epithelial cells, macrophages, and dendritic cells), adverse effects were also only found at high silver concentrations. The silver ions that are released from silver nanoparticles may be harmful to skin with disrupted barrier (e.g., wounds) and induce oxidative stress in skin cells (HaCaT). In conclusion, the data obtained on the effects of this well-defined type of silver nanoparticles on various biological systems clearly demonstrate that cell-type specific properties as well as experimental conditions determine the biocompatibility of and the cellular responses to an exposure with silver nanoparticles.

In vitro interaction of colloidal nanoparticles with mammalian cells: What have we learned thus far?

The interfacing of colloidal nanoparticles with mammalian cells is now well into its second decade. In this review our goal is to highlight the more generally accepted concepts that we have gleaned from nearly twenty years of research. While details of these complex interactions strongly depend, amongst others, upon the specific properties of the nanoparticles used, the cell type, and their environmental conditions, a number of fundamental principles exist, which are outlined in this review.

Deliquescence and efflorescence behavior of ternary inorganic/organic/water aerosol particles.

The deliquescence behavior of ternary inorganic (ammonium sulfate and ammonium nitrate)/organic (glutaric acid and malonic acid)/water aerosol particles has been investigated at 293 K using a novel surface aerosol microscopy (SAM) technique. The results obtained for the deliquescence relative humidities (DRH) for particles of variable inorganic/organic contents show a eutectic behavior with the mixed particles showing deliquescence at lower DRH compared to the pure inorganic and organic components, respectively. This behavior has been quantitatively modeled using the extended aerosol inorganics (E-AIM) thermodynamic model of Clegg et al. in combination with the UNIFAC group activity approach to account for organic molecular solutes. In addition, we have investigated the crystallization behavior of supersatured and formerly deliquesced ternary solution droplets using space resolved Raman spectroscopy. It is found that such droplets produce solid particles in which the inorganic and organic phases show some spatial separation with the organic component being predominantly found at the outer part of the particle. Independent measurements of the contact angles of such ternary droplets reveal that their angles are within experimental error identical to those of the purely organic/water solutions.

Nanoparticle size is a critical physicochemical determinant of the human blood plasma corona: a comprehensive quantitative proteomic analysis.

In biological fluids, proteins associate with nanoparticles, leading to a protein "corona" defining the biological identity of the particle. However, a comprehensive knowledge of particle-guided protein fingerprints and their dependence on nanomaterial properties is incomplete. We studied the long-lived ("hard") blood plasma derived corona on monodispersed amorphous silica nanoparticles differing in size (20, 30, and 100 nm). Employing label-free liquid chromatography mass spectrometry, one- and two-dimensional gel electrophoresis, and immunoblotting the composition of the protein corona was analyzed not only qualitatively but also quantitatively. Detected proteins were bioinformatically classified according to their physicochemical and biological properties. Binding of the 125 identified proteins did not simply reflect their relative abundance in the plasma but revealed an enrichment of specific lipoproteins as well as proteins involved in coagulation and the complement pathway. In contrast, immunoglobulins and acute phase response proteins displayed a lower affinity for the particles. Protein decoration of the negatively charged particles did not correlate with protein size or charge, demonstrating that electrostatic effects alone are not the major driving force regulating the nanoparticle-protein interaction. Remarkably, even differences in particle size of only 10 nm significantly determined the nanoparticle corona, although no clear correlation with particle surface volume, protein size, or charge was evident. Particle size quantitatively influenced the particle's decoration with 37% of all identified proteins, including (patho)biologically relevant candidates. We demonstrate the complexity of the plasma corona and its still unresolved physicochemical regulation, which need to be considered in nanobioscience in the future.

Spatial separation of individual substances in effloresced crystals of ternary ammonium sulphate/dicarboxylic acid/water aerosols.

This work examines the crystals resulting from the efflorescence of internally mixed aqueous aerosols comprising ammonium sulphate and different dicarboxylic acids. Most studies on the deliquescence of aerosols use previously effloresced aerosols in their experiments. However, during efflorescence a highly supersaturated solution crystallises in a kinetically controlled way unlike the case of thermodynamically controlled crystallisation. Herein the distribution of individual substances within the effloresced crystals is investigated using Raman scanning experiments. The data presented show an intriguingly complex behaviour of these ternary and quarternary aerosols. A spatial separation of substances in the crystals resulting from the efflorescence of previously internally mixed ternary salt/dicarboxylic acid/water aerosol droplets is demonstrated and mechanistic aspects are discussed.

The influence of surface composition of nanoparticles on their interactions with serum albumin.

Interactions between differently functionalised silver and gold nanoparticles (NPs) as well as polystyrene nanoparticles with bovine serum albumin (BSA) are studied using circular dichroism (CD) spectroscopy. It is found that the addition of NPs to the protein solution destroys part of the helical secondary structure of the protein as a result of surface adsorption. From the loss of free protein and hence the extent of their structural change adsorption equilibrium constants are derived. The results reveal that citrate-coated gold and silver NPs exhibit much stronger interactions with BSA than polymeric or polymer-coated metallic NPs. It is therefore concluded that for the particles considered, the influence of surface composition on the interaction behaviour dominates that of the core.

Collisions of noble gases with supercooled sulfuric acid-water solutions.

The collisions of hyperthermal noble gases (He, Ne, Ar, Kr, Xe) with supercooled binary sulfuric acid-water mixtures (57-77 wt%) were explored in the temperature range between 210 and 240 K. The experiments were performed by directing a molecular beam of the respective gases onto a continuously renewed liquid surface and monitoring the velocity of the scattered molecules by mass spectrometry. Depending on the initial translational energies and molecular masses, we observe both inelastic scattering from the surface as well as thermalization followed by subsequent desorption. The experiments indicate that the repulsive momentum transfer in the inelastic scattering channel increases with increasing mass of the impinging gas, while it is only weakly affected by the initial velocities. The final energy of the thermally desorbing atoms can always be approximated by a Maxwell-Boltzmann distribution equal to the liquid bulk phase temperature. The influence of the binary composition of the liquid phase is only noticeable in the case of Ne, whilst this dependence diminishes for gases with molecular masses >or=40 amu. The probability of thermalisation relative to inelastic scattering increases with the bulk phase temperature, independent of the molecular masses of the colliding gas. In contrast, the fractional energy transfer during collision does not increase with temperature, except for Neon. These results can be interpreted in the model framework of hard-sphere collisions of noble gases with the surface, during which water and sulfuric acid molecules interact independently with the impinging gas.

Water uptake on mineral dust and soot: a fundamental view of the hydrophilicity of atmospheric particles?

The interaction of water vapour with mineral dust and soot surfaces has been studied in the temperature range 203 K < T < 298 K using a Knudsen cell reactor. For the uptake of water on mineral dust an initial uptake coefficient of gamma(ini) = (6.3 +/- 0.7) x 10(-2) independent of temperature has been determined. In contrast the desorption rate has been found to be strongly temperature dependent with desorption rate constants decreasing from 1 x 10(-3) at 265 K to 1 x 10(-4) at 223 K. In addition, relatively high surface coverages have been determined from which an adsorption enthalpy of -40 kJ mol(-1) is inferred. For the uptake of water on soot the initial uptake coefficient has been found to be independent of temperature with a value of gamma(ini) = (4.7 +/- 0.2) x 10(-2), similar to the case of mineral dust. However, the corresponding desorption rate constants have been found to be three orders of magnitude larger than for mineral dust. Consistent with this finding, low surface coverages with an adsorption enthalpy of -27 kJ mol(-1) have been derived. A comparison of the uptake kinetics and adsorption enthalpies of water on mineral dust and soot leads to the conclusion that water is much stronger interacting with mineral dust than with soot. In terms of a hydrophilicity concept the results suggest, that mineral dust may be regarded as hydrophilic whereas soot is hydrophobic and that fundamental kinetic and thermochemical quantities may be related to that concept.