Publikationen

Abstract Streptococcus pneumoniae infections are a leading cause of death worldwide. Bacterial membrane vesicles (MVs) are promising vaccine candidates because of the antigenic components of their parent microorganisms. Pneumococcal MVs exhibit low toxicity towards several cell lines, but their clinical translation requires a high yield and strong immunogenic effects without compromising immune cell viability. MVs are isolated during either the stationary phase (24 h) or death phase (48 h), and their yields, immunogenicity and cytotoxicity in human primary macrophages and dendritic cells have been investigated. Death-phase vesicles showed higher yields than stationary-phase vesicles. Both vesicle types displayed acceptable compatibility with primary immune cells and several cell lines. Both vesicle types showed comparable uptake and enhanced release of the inflammatory cytokines, tumor necrosis factor and interleukin-6, from human primary immune cells. Proteomic analysis revealed similarities in vesicular immunogenic proteins such as pneumolysin, pneumococcal surface protein A, and IgA1 protease in both vesicle types, but stationary-phase MVs showed significantly lower autolysin levels than death-phase MVs. Although death-phase vesicles produced higher yields, they lacked superiority to stationary-phase vesicles as vaccine candidates owing to their similar antigenic protein cargo and comparable uptake into primary human immune cells.

The family of sputter deposited granular metal-based carbon-containing sensor films is known for their high sensitivity transforming force-dependent strain into electrical resistance change. Among them nickel–carbon thin films possess a gauge factor of up to 30, compared to only 2 for traditional sensor films of metal alloys. This high sensitivity is based on disordered interparticle tunneling through barriers of graphite-like carbon walls between metal–carbon particles of columnar shape. Force and pressure sensors would benefit a lot from the elevated piezoresistivity. A disadvantage, however, is a disturbing temporal creep and drift of the resistance under load and temperature. This contribution shows how to stabilize such sensor films. A significant stabilization is achieved by partially replacing nickel with chromium, albeit at the expense of sensitivity. The more chromium used in these NixCr1−x-C layers, the higher the optimum annealing temperature can be selected and the better the electrical stabilization. A good compromise while maintaining sensitivities well above the standard of 2 is identified for films with x=0.5 to 0.9, stabilized by optimized temperature treatments. The stabilizing effect of chromium is revealed by transmission electron microscopy with elemental analysis. The post-annealing drives segregation processes in the layer material. While the interior of the layer is depleted of chromium and carbon, boundary layers are formed. Chromium is enriched near the surface boundary, oxidized in air and forms chromium-rich oxide sub-layers, which are chemically very stable and protect against further reactions and corrosion. As a result, creep and drift errors are greatly reduced, so that the optimized sensor coatings are now suitable for widespread use.

Glioblastoma is the most common malignant brain tumor with a high grade of recurrence, invasiveness, and aggressiveness. Currently, there are no curative treatments; therefore, the discovery of novel molecules with anti-tumor activity or suitable drug delivery systems are important research topics. The aim of the present study was to investigate the anti-tumor activity of Givinostat, a pan-HDAC inhibitor, and to design an appropriate liposomal formulation to improve its pharmacokinetics profile and brain delivery. The present work demonstrates that the incorporation of Givinostat in liposomes composed of cholesterol and sphingomyelin improves its in vivo half-life and increases the amount of drug reaching the brain in a mouse model. Furthermore, this formulation preserves the anti-tumor activity of glioblastoma in 2D and 3D in vitro models. These features make liposome-Givinostat formulations potential candidates for glioblastoma therapy.

Abstract Extracellular vesicles (EV) are an emerging technology as immune therapeutics and drug delivery vehicles. However, EVs are usually stored at −80 °C which limits potential clinical applicability. Freeze-drying of EVs striving for long-term stable formulations is therefore studied. The most appropriate formulation parameters are identified in freeze-thawing studies with two different EV types. After a freeze-drying feasibility study, four lyophilized EV formulations are tested for storage stability for up to 6 months. Freeze-thawing studies revealed improved colloidal EV stability in presence of sucrose or potassium phosphate buffer instead of sodium phosphate buffer or phosphate-buffered saline. Less aggregation and/or vesicle fusion occurred at neutral pH compared to slightly acidic or alkaline pH. EVs colloidal stability can be most effectively preserved by addition of low amounts of poloxamer 188. Polyvinyl pyrrolidone failed to preserve EVs upon freeze-drying. Particle size and concentration of EVs are retained over 6 months at 40 °C in lyophilizates containing 10 mm K- or Na-phosphate buffer, 0.02% poloxamer 188, and 5% sucrose. The biological activity of associated beta-glucuronidase is maintained for 1 month, but decreased after 6 months. Here optimized parameters for lyophilization of EVs that contribute to generate long-term stable EV formulations are presented.

Nanofabrication technologies have been recently applied to the development of engineered nano–bio interfaces for manipulating complex cellular processes. In particular, vertically configurated nanostructures such as nanoneedles (NNs) have been adopted for a variety of biological applications such as mechanotransduction, biosensing, and intracellular delivery. Despite their success in delivering a diverse range of biomolecules into cells, the mechanisms for NN-mediated cargo transport remain to be elucidated. Recent studies have suggested that cytoskeletal elements are involved in generating a tight and functional cell–NN interface that can influence cargo delivery. In this study, by inhibiting actin dynamics using two drugs—cytochalasin D (Cyto D) and jasplakinolide (Jas), we demonstrate that the actin cytoskeleton plays an important role in mRNA delivery mediated by silicon nanotubes (SiNTs). Specifically, actin inhibition 12 h before SiNT-cellular interfacing (pre-interface treatment) significantly dampens mRNA delivery (with efficiencies dropping to 17.2% for Cyto D and 33.1% for Jas) into mouse fibroblast GPE86 cells, compared to that of untreated controls (86.9%). However, actin inhibition initiated 2 h after the establishment of GPE86 cell–SiNT interface (post-interface treatment), has negligible impact on mRNA transfection, maintaining > 80% efficiency for both Cyto D and Jas treatment groups. The results contribute to understanding potential mechanisms involved in NN-mediated intracellular delivery, providing insights into strategic design of cell–nano interfacing under temporal control for improved effectiveness.

Back-contact electrodes for hybrid organic-inorganic perovskite solar cells (PSCs) eliminate the parasitic absorption losses caused by the transparent conductive electrodes that are inherent to conventional sandwich-architecture devices. However, the fabrication methods for these unconventional architectures rely heavily on expensive photolithography, which limits scalability. Herein, we present an alternative cost-effective microfabrication technique in which the conventional photolithography process is replaced by microsphere lithography in which a close-packed polystyrene microsphere monolayer acts as the patterning mask for the honeycomb-shaped electrodes. A comprehensive comparison between photolithography and microsphere lithography fabrication techniques was conducted. Using microsphere lithography, we achieve highly efficient devices having a stabilized power conversion efficiency (PCE) of 8.6%, twice the reported value using photolithography. Microsphere lithography also enabled the fabrication of the largest back-contact PSC to date, having an active area of 0.75 cm2 and a stabilized PCE of 2.44%.

Living organisms are open systems that can incorporate externally provided nutrients to vary their appearances and properties, while synthetic materials normally have fixed sizes, shapes, and functions. Herein, we report a strategy for enabling cross-linked polymers to continuously grow with programmable bulky structures and properties. The growing strategy involves repeatable processes including swelling of polymerizable components into the cross-linked polymers, in situ polymerization of the components, and homogenization of the original and newborn polymer networks. Using acrylate-based polymers as an example, we demonstrate that homogenization allows the grown polymer materials to further integrate various polymerizable components to alternate their bulky properties. During the growth, the changes from elastomers to organogels and then to hydrogels with updated covalent-linked functions (i.e., photochromism and thermoresponsiveness) are shown. Since this growing strategy is applicable to different acrylate systems, we envision its great potential in the design of next-generation polymers, smartening systems, and postmodification of cross-linked polymer materials.

Abstract Tunable vertically aligned nanostructures, usually fabricated using inorganic materials, are powerful nanoscale tools for advanced cellular manipulation. However, nanoscale precision typically requires advanced nanofabrication machinery and involves high manufacturing costs. By contrast, polymeric nanoneedles (NNs) of precise geometry can be produced by replica molding or nanoimprint lithography—rapid, simple, and cost-effective. Here, cytocompatible polymeric arrays of NNs are engineered with identical topographies but differing stiffness, using polystyrene (PS), SU8, and polydimethylsiloxane (PDMS). By interfacing the polymeric NN arrays with adherent and suspension mammalian cells, and comparing the cellular responses of each of the three polymeric substrates, the influence of substrate stiffness from topography on cell behavior is decoupled. Notably, the ability of PS, SU8, and PDMS NNs is demonstrated to facilitate mRNA delivery to GPE86 cells with 26.8% ± 3.5%, 33.2% ± 7.4%, and 30.1% ± 4.1% average transfection efficiencies, respectively. Electron microscopy reveals the intricacy of the cell–NN interactions; and immunofluorescence imaging demonstrates that enhanced endocytosis is one of the mechanisms of PS NN-mediated intracellular delivery, involving the endocytic proteins caveolin-1 and clathrin heavy chain. The results provide insights into the interfacial interactions between cells and polymeric NNs, and their related intracellular delivery mechanisms.

Cononsolvency is an intriguing phenomenon where a polymer collapses in a mixture of good solvents. This cosolvent-induced modulation of the polymer solubility has been observed in solutions of several polymers and biomacromolecules, and finds application in areas such as hydrogel actuators, drug delivery, compound detection and catalysis. In the past decade, there has been a renewed interest in understanding the molecular mechanisms which drive cononsolvency with a predominant emphasis on its connection to the preferential adsorption of the cosolvent. Significant efforts have also been made to understand cononsolvency in complex systems such as micelles, block copolymers and thin films. In this review, we will discuss some of the recent developments from the experimental, simulation and theoretical fronts, and provide an outlook on the problems and challenges which are yet to be addressed.

Due to the limited available amounts of components, especially of low water-soluble drugs, formulation development is often impeded by a careful characterization. The use of small batch sizes might solve this problem but requires also adequate analytics. Concentration of nanoparticulate formulations lack straightforward evaluation techniques. In this work, a precise and straight-forward method is established to individually count nanoparticles. A microfluidic chip with known dimensions was used to visualize single particles flowing through the channel (single-particle tracking (SPT)). A sequence of 10,000 images was analyzed to determine the mean particle concentration. The proposed method is independent of the particular flow rate through the microfluidic chip as long as there is no particle overlap and a continuous exchange of particles. Monodisperse Rhodamine B labeled poly (methyl methacrylate) (PMMA) nanoparticles (267.03 ± 9.79 nm) were used as a model and reference particle system for the evaluation process of SPT allowing for a gravimetric determination based on density analysis using analytical ultracentrifugation (AUC) and gas pycnometry. The SPT method was evaluated and compared to other techniques used for concentration measurement. Both approaches (SPT and gravimetry) provide very similar and comparable results indicating the applicability of this novel quantification approach. In contrast, multi angle dynamic light scattering (MADLS) could not yield a precision as good as SPT (number density rel. standard deviation SD nSPT = 11.67%; SD nMADLS = 49.45%). Finally, the measured particle number concentrations can be realized in low concentration ranges (0.8249 μg mL−1 – 0.08249 μg mL−1) not accessible for MADLS (0.08249 mg mL−1 – 0.008249 mg mL−1) and gravimetric analysis.