Datei im Netzwerk der BAM verfügbar ("Closed Access")
Filtern
Dokumenttyp
- Zeitschriftenartikel (11)
- Buchkapitel (1)
- Beitrag zu einem Sammelband (1)
Schlagworte
Drug and gene delivery via nanoparticles across biological barriers such as the alveolar-capillary barrier of the lung constitutes an interesting and increasingly relevant field in nanomedicine. Nevertheless, potential hazardous effects of nanoparticles (NPs) as well as their cellular and systemic fate should be thoroughly examined. Hence, this study was designed to evaluate the effects of amorphous silica NPs (Sicastar) and (poly)organosiloxane NPs (AmOrSil) on the viability and the inflammatory response as well as on the cellular uptake mechanisms and fate in cells of the alveolar barrier. For this purpose, the alveolar epithelial cell line (NCI H441) and microvascular endothelial cell line (ISO-HAS-1) were used in an experimental set up resembling the alveolar-capillary barrier of the lung. In terms of IL-8 and sICAM Sicastar resulted in harmful effects at higher concentrations (60 µg/ml) in conventional monocultures but not in the coculture, whereas AmOrSil showed no significant effects. Immunofluorescence counterstaining of endosomal structures in NP-incubated cells showed no evidence for a clathrin- or caveolae-mediated uptake mechanism. However, NPs were enclosed in flotillin-1 and -2 marked vesicles in both cell types. Flotillins appear to play a role in cellular uptake or trafficking mechanisms of NPs and are discussed as indicators for clathrin- or caveolae-independent uptake mechanisms. In addition, we examined the transport of NPs across this in vitro model of the alveolar-capillary barrier forming a tight barrier with a transepithelial electrical resistance of 560 ± 8 Ω cm². H441 in coculture with endothelial cells took up much less NPs compared to monocultures. Moreover, coculturing prevented the transport of NP from the epithelial compartment to the endothelial layer on the bottom of the filter insert. This supports the relevance of coculture models, which favour a differentiated and polarised epithelial layer as in vitro test systems for nanoparticle uptake.
Interaction forces between all objects are either of repulsive or attractive nature. Concerning attractive interactions, the determination of dispersion forces are of special interest since they appear in all colloidal systems and have a crucial influence on the properties and processes in these systems. One possibility to link theory and experiment is the description of the London–Van der Waals forces in terms of the Hamaker constant, which leads to the challenging problem of calculating the van der Waals interaction energies between colloidal particles. Hence, the determination of a Hamaker constant for a given material is needed when interfacial phenomena such as adhesion are discussed in terms of the total potential energy between particles and substrates. In this work, the asymmetrical flow field-flow fractionation (AF-FFF) in combination with a Newton algorithm based iteration process was used for the determination of Hamaker constants of different nanoparticles in toluene.
Cylindrical poly(styrene sulfonate) brushes were combined with a tetravalent double-DABCO based counterion. With increasing addition of counterion, hydrodynamic radius and radius of gyration decrease and AFM reveals a conformation change from wormlike to curled structures. Rg/Rh changes from 1.3 to 0.9. Above charge stoichiometry, brushes become interconnected into finite size assemblies, which are stable in aqueous solution for charge ratios (ratio of the molar concentration of charged counterion groups to the molar concentration of charged monomer units of the polymer brush) 1 < Ι < 1.2. They are compact and consist of the brushes in curled (bent) conformation, which is in difference to network-like aggregates from cylindrical poly(styrene sulfonate) brushes with tetravalent porphyrin counterions (Gröhn et al. Macromolecules 2009) and to a wormlike — bent conformation — helix transition found for another brush counterion system (Müller et al. Soft Matter 2009).
Continuous-flow process conditions for the reduction to gold nanoparticles were varied following a variation scheme aiming at impacting each of the elementary steps–mixing, reaction, polymer diffusion and polymer binding. While the last three variations are done by proper material choice (reducer; polymeric stabilizer), flow processing using a multilamination micromixer intensified the mass transfer in all these steps and particularly improved the first mixing step of the reactants. As reducers L-ascorbic acid (H2Asc) and sodium borohydride (NaBH4) were used and as gold particle precursor chloroauric acid (HAuCl4). All processing runs were analysed by plasmon absorption and transmission electron microscopy to reveal size, shape and polydispersity. For the HAuCl4-H2Asc system, the smallest particles were 0.84 nm (7.5 mL/min) and 0.63 nm (10.0 mL/min) for the HAuCl4-NaBH4 system. This is at the upper edge of performance reported for batch systems in literature, i.e. among the smallest gold particles generated. While the particles made for the HAuCl4-H2Asc system were polydisperse and polyshaped for most process conditions (however being spherical and relatively uniform at the two largest flow rates investigated), processing with the HAuCl4-NaBH4 system resulted in particles sizes with small mean deviation and almost ideal spherical shape. With polyvinyl alcohol (PVA) as stabilizer, stable colloids were obtained for one week (violet coloured) up to at least 2 months (pink coloured) in case of reaction with ascorbic acid, and for at least 1 month in case of reaction with NaBH4.
Polystyrene sulfonate - porphyrin assemblies: influence of polyelectrolyte and porphyrin structure
(2011)
In this study, electrostatic self-assembly of different polystyrene sulfonates and a set of tetravalent cationic porphyrins is investigated. It is shown that association of linear polystyrene sulfonates of different molar masses yields finite size nanoscale assemblies that are stable in aqueous solution. Aggregates are compared to the ones of cylindrical brushes, revealing that both form assemblies in the 100 nm range with the charge ratio (molar ratio of porphyrin charges to polyelectrolyte charges) being determining, while the morphology of the resulting network-like assemblies is different for both polyelectrolyte architectures. For the smallest 8k polystyrene sulfonate, in addition, stoichiometric conditions differ. The influence of the molecular porphyrin structure was investigated by comparing meso-tetrakis(4-(trimethyl-ammonium)phenyl)porphyrin (TAPP) with its Cu(II) and Zn(II) loaded analogues and meso-tetrakis(4-N-methylpyridinium)porphyrin (TMPyP), revealing differences in stacking tendency and geometry. Additionally, the TMPyP accumulates more in the inside of the brush than the other porphyrins, likely due to the different position of its charged groups. The supramolecular nanostructures formed were characterized by UV–vis spectroscopy, light scattering, atomic force microscopy, cryo transmission electron microscopy, and small-angle neutron scattering. Results may build a valuable basis for the use of polyelectrolyte–porphyrin assemblies in medicine, catalysis, or energy conversion.
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 particles 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.
We present the synthesis and characterization of multifunctional fluorophore-labeled poly(organosiloxane) nanoparticles with core-shell architecture, where the fluorescent dye is incorporated into the core. Grafting of heterobifunctional poly(ethylene oxide) (PEO) onto the particle surface leads to water-soluble biocompatible nanoparticles. Two different strategies have been used for the synthesis: The encapsulation of dye-labeled monomers during the polycondensation with additional PEO coating and subsequent dye labeling by covalent attachment of the fluorescent dye rhodamine B to the (chloromethylphenyl)siloxane groups in the core after polymerization and grafting of PEO onto the surface. Comparison of the fluorescence quantum yields of the nanoparticles before and after PEO coating show a decrease in quantum yield after PEO coating.
Developing polymersomes for drug delivery purposes requires a deeper understanding of their behavior in physiological environment. We performed the self-assembly and in-situ loading of polybutadiene-block-polyethyleneoxide (PB-b-PEO) polymersomes in a continuous process using micromixers. Varying the length and end-groups of the starting block copolymer allows us to control the polymer membrane thickness and surface functionalities (hydroxyl or carboxylic acid), required to realize a further coupling with specific cell targeting ligands. To get a deeper understanding of these polymersomes in physiological environment, we studied the cellular response (HeLa cells) in presence of various polymersomes, and showed by cytotoxicity tests the relative biocompatibility of the systems. Flow cytometry experiments at 4?°C in PBS buffer showed a different behavior of hydroxyl-functionalized vesicles compared to carboxylic acid-functionalized vesicles. On the contrary cell binding in DMEM medium supplemented with 10% FCS was almost completely blocked with both kinds of polymersomes. Protein adsorption measurements by dynamic light scattering confirmed that protein binding occurs in all cases, which apparently influences the particle-cell interaction. This study contributes towards a deeper understanding of polymersomes in biological environment and further investigations will help us to design highly effective polymersomes for in vitro as well as in vivo applications.
Synthetic molecular systems undergoing structure and property changes as a response to external stimuli offer a series of relevant functions. Their direct use in solution, immobilized at surfaces, or integrated into adequate matrices, allow for the elaboration of nano-carriers, molecular electronic memory circuits, adaptive systems, artificial muscles, or healable materials, among others. Reversible systems are commonly generated by utilizing molecular switches as responsive bi-stable molecules. Of particular interest are switches controlled by non-invasive triggers, especially light, which can display high spatial and temporal resolution. Photoswitches were used to change properties, such as conductance and basicity, based on light-induced alteration of the molecular geometry, that is, their dimensions and shape. In nature, light-induced geometrical modifications of molecular systems lead to primary biological functions, such as the visual perception in vertebrates, or proton pumping in some bacteria, two examples that nicely illustrate the potential of photo-inducing structural modifications in organic systems.
Poly(butadiene)-b-poly(ethylene oxide) vesicles were successfully loaded with hydrophilic dye Phloxine B. Dye addition during vesicle formation leads to Phloxine B encapsulated inside the water filled vesicle core as well as to freely diffusing dye molecules. The removal of uncapsulated substrate involves time consuming methods like dialysis or harsher methods like ultra filtration or selective precipitation, posing the risk of irreversible sample manipulation. Here used Phloxine B as pH sensitive fluorescence indicator allows the characterization of hydrophilic loading without separation procedure by adjusting the pH value. Additionally membrane blocking efficiency can be studied by time dependent fluorescence measurements. Cryogenic TEM studies showed that the self-assembled structure remained unchanged when the hydrophilic dye was incorporated within the vesicles. Fluorescence microscopy imaging proved the encapsulation of the hydrophilic dye inside the core volume. The combination of fluorescence correlation spectroscopy (FCS) and dynamic light scattering (DLS) measurements as ensemble methods confirmed those results additionally.