Publikationen

Titanium dioxide nanoparticles (TiO2 NPs) have a wide range of applications in several industrial and biomedical domains. Based on the evidence, the workers exposed to inhaled nanosized TiO2 powder are more susceptible to the risks of developing respiratory diseases. Accordingly, this issue has increasingly attracted the researchers’ interest in understanding the consequences of TiO2 NPs exposure. Regarding this, the present study was conducted to analyze the local effects of TiO2 NPs on allergic airway inflammation and their uptake in a mouse model of ovalbumin (OVA)-induced allergic airway inflammation.

Aluminum Nitride (AlN) is a well-known compound piezoelectric material with high Complementary Metal-Oxide Semiconductor process compatibility. Previous results show that the piezoelectric coefficient correlates with the c-axis orientation of AlN. In this work, the optical properties of reactively sputtered AlN thin-films have been investigated to find a relation with the structural properties of the film (i.e. c-axis orientation) using spectroscopic ellipsometry. The results show that for almost the same film thickness, highly c-axis orientated films have higher refractive indices compared to films with low c-axis orientation or amorphous layers. A relationship between (002) peak intensity measured with x-ray diffractometry and refractive index is shown. At 546 nm wavelength, the refractive index decreased from 2.15 for films with high (002) peak intensity, to 2.11 for films with about half the peak intensity, and further down to 1.90 for films without preferred orientation. Additionally, with the same sputtering conditions, the variation of the film thickness seems to have no significant effect on the refractive index. The results of AlN thin-film mass density obtained from x-ray reflectivity measurements are consistent with refractive index measurements. The mass density of AlN thin-films with high c-axis orientation that resulted in a higher refractive index is 2.99 g/cm−3, compared to 3.23 g/cm−3 for epitaxial grown AlN layers. This value decreased to 2.82 g/cm−3 for films with poor c-axis orientation.

The interaction of cyclopeptides bound to the surface of mixed monolayer-protected gold nanoparticles with sulfate anions causes the crosslinking and concomitant precipitation of the nanoparticles from aqueous solutions even in presence of an excess of competing anions, thus allowing the naked eye detection of sulfate in water.

In 2019, it was estimated that 2.5 million people die from lower tract respiratory infections annually. One of the main causes of these infections is Staphylococcus aureus, a bacterium that can invade and survive within mammalian cells. S. aureus intracellular infections are difficult to treat because several classes of antibiotics are unable to permeate through the cell wall and reach the pathogen. This condition increases the need for new therapeutic avenues, able to deliver antibiotics efficiently. In this work, we obtained outer membrane vesicles (OMVs) derived from the myxobacteria Cystobacter velatus strain Cbv34 and Cystobacter ferrugineus strain Cbfe23, that are naturally antimicrobial, to target intracellular infections, and investigated how they can affect the viability of epithelial and macrophage cell lines. We evaluated by cytometric bead array whether they induce the expression of proinflammatory cytokines in blood immune cells. Using confocal laser scanning microscopy and flow cytometry, we also investigated their interaction and uptake into mammalian cells. Finally, we studied the effect of OMVs on planktonic and intracellular S. aureus. We found that while Cbv34 OMVs were not cytotoxic to cells at any concentration tested, Cbfe23 OMVs affected the viability of macrophages, leading to a 50% decrease at a concentration of 125,000 OMVs/cell. We observed only little to moderate stimulation of release of TNF-alpha, IL-8, IL-6 and IL-1beta by both OMVs. Cbfe23 OMVs have better interaction with the cells than Cbv34 OMVs, being taken up faster by them, but both seem to remain mostly on the cell surface after 24 h of incubation. This, however, did not impair their bacteriostatic activity against intracellular S. aureus. In this study, we provide an important basis for implementing OMVs in the treatment of intracellular infections.

Abstract There is a lack of efficient therapies to treat increasingly prevalent autoimmune diseases, such as inflammatory bowel disease and celiac disease. Membrane vesicles (MVs) isolated from probiotic bacteria have shown tremendous potential for treating intestinal inflammatory diseases. However, possible dilution effects and rapid elimination in the gastrointestinal tract may impair their application. A cell-free and anti-inflammatory therapeutic system?probiomimetics?based on MVs of probiotic bacteria (Lactobacillus casei and Lactobacillus plantarum) coupled to the surface of microparticles is developed. The MVs are isolated and characterized for size and protein content. MV morphology is determined using cryoelectron microscopy and is reported for the first time in this study. MVs are nontoxic against macrophage-like dTHP-1 and enterocyte-like Caco-2 cell lines. Subsequently, the MVs are coupled onto the surface of microparticles according to facile aldehyde-group functionalization to obtain probiomimetics. A significant reduction in proinflammatory TNF-α level (by 86%) is observed with probiomimetics but not with native MVs. Moreover, it is demonstrated that probiomimetics have the ability to ameliorate inflammation-induced loss of intestinal barrier function, indicating their potential for further development into an anti-inflammatory formulation. These engineered simple probiomimetics that elicit striking anti-inflammatory effects are a key step toward therapeutic MV translation.

Polymerization of carbazole dicarboxaldehydes and lysine derivatives by imine and acylhydrazone formation afforded peptide-derived molecular biodynamers. Characterization of their physicochemical properties revealed an interesting morphology change upon polymerization from monomers forming submicrometer spherical micelles to nanometer-sized rigid-rod-shaped polymeric particles. A combination of light-scattering methods, small-angle neutron scattering, and transmission electron microscopy enabled a detailed investigation of this morphological change. Moreover, we investigated by dynamic and static light scattering how the pH affects the fluorescence and size of the biodynamers. These morphological and pH-dependent changes are expected to open the door to a myriad of applications of molecular biodynamers.

Gold nanoparticles covered with a mixture of ligands of which one type contains solubilizing triethylene glycol residues and the other peripheral zinc(II)–dipicolylamine (DPA) complexes allowed the optical detection of hydrogenphosphate, diphosphate, and triphosphate anions in water/methanol 1:2 (v/v). These anions caused the bright red solutions of the nanoparticles to change their color because of nanoparticle aggregation followed by precipitation, whereas halides or oxoanions such as sulfate, nitrate, or carbonate produced no effect. The sensitivity of phosphate sensing depended on the nature of the anion, with diphosphate and triphosphate inducing visual changes at significantly lower concentrations than hydrogenphosphate. In addition, the sensing sensitivity was also affected by the ratio of the ligands on the nanoparticle surface, decreasing as the number of immobilized zinc(II)–dipicolylamine groups increased. A nanoparticle containing a 9:1 ratio of the solubilizing and the anion-binding ligand showed a color change at diphosphate and triphosphate concentrations as low as 10 μmol/L, for example, and precipitated at slightly higher concentrations. Hydrogenphosphate induced a nanoparticle precipitation only at a concentration of ca. 400 μmol/L, at which the precipitates formed in the presence of diphosphates and triphosphates redissolved. A nanoparticle containing fewer binding sites was more sensitive, while increasing the relative number of zinc(II)–dipicolylamine complexes beyond 25% had a negative impact on the limit of detection and the optical response. Transmission electron microscopy provided evidence that the changes of the nanoparticle properties observed in the presence of the phosphates were due to a nanoparticle crosslinking, consistent with the preferred binding mode of zinc(II)–dipicolylamine complexes with phosphate anions which involves binding of the anion between two metal centers. This work thus provided information on how the behavior of mixed monolayer-protected gold nanoparticles is affected by multivalent interactions, at the same time introducing a method to assess whether certain biologically relevant anions are present in an aqueous solution within a specific concentration range.

In recent years, extracellular vesicles (EVs) and outer membrane vesicles (OMVs) have become an extensive and diverse field of research. They hold potential as diagnostic markers, therapeutics and for fundamental biological understanding. Despite ongoing studies, numerous information regarding function, content and stability of EVs remains unclear. If EVs and OMVs ought to be used as therapeutics and in clinical environments, their stability is one of the most important factors to be considered. Especially for formulation development, EVs and OMVs need to be stable at higher temperatures. To the best of our knowledge, very little work has been published regarding heat stability of neither EVs nor OMVs. In the present study, we investigated B lymphoblastoid cell-derived EVs and OMVs derived from myxobacterial species Sorangiineae as model vesicles. We exposed the vesicles to 37 °C, 50 °C, 70 °C and 100 °C for 1 h, 6 h and 24 h, and also autoclaved them. Interestingly, physico-chemical analyses such as size, particle concentration and protein concentration showed minor alterations, particularly at 37 °C. Flow cytometry analysis emphasised these results suggesting that after heat impact, EVs and OMVs were still able to be taken up by macrophage-like dTHP-1 cells. These data indicate that both mammalian and bacterial vesicles show intrinsic stability at physiological temperature. Our findings are important to consider for vesicle formulation and for advanced bioengineering approaches.

A novel approach for electrochemical tuning of alcohol oxidase (AOx) and alcohol dehydrogenase (ADH) biocatalysis towards butanol-1 oxidation by incorporating enzymes in various designs of amperometric biosensors is presented. The biosensors were developed by using commercial graphene oxide-based screen-printed electrodes and varying enzyme producing strains, encapsulation approaches (layer-by-layer (LbL) or one-step electrodeposition (EcD)), layers composition and structure, operating conditions (applied potential values) and introducing mediators (Meldola Blue and Prussian Blue) or Pd-nanoparticles (Pd-NPs). Simultaneous analysis/screening of multiple crucial system parameters during the enzyme engineering process allowed to identify within a period of one month that four out of twelve proposed designs demonstrated a good signal reproducibility and linear response (up to 14.6 mM of butanol) under very low applied potentials (from −0.02 to −0.32 V). Their mechanical stability was thoroughly investigated by multi-analytical techniques prior to butanol determination in cell-free samples from an anaerobic butanol fermentation. The EcD-based biosensor that incorporates ADH, NAD+, Pd-NPs and Nafion showed no loss of enzyme activity after preparation and demonstrated capabilities towards low potential (−0.12 V) detection of butanol-1 in fermentation medium (4 mM) containing multiple electroactive species with almost 15 times enhanced sensitivity (0.2282 μA/mM ± 0.05) when compared to the LbL design. Furthermore, the ADH-Nafion bonding for the S. cerevisiae strain was confirmed to be 3 times higher than for E. coli.

Pore collapse upon drying is a well-known phenomenon in ultrafiltration PES membranes. Here we demonstrate how alteration of membrane surface chemistry can be used to control the extent of pore collapse and ultimately to tailor membrane properties. Commercial hollow-fiber PES membranes were modified via surface-initiated ATRP to obtain different polymer-grafted membranes and were subsequently dried to facilitate pore collapse. The different polymer grafts could be used for controlling the water flux and solute rejection characteristics of the membranes. Controlled membrane pore collapse could be exploited to obtain higher rejection of sodium chloride, magnesium sulfate and calcein. Calcein as the largest solute showed almost full rejection (98.9 ± 0.3%) on the membrane. The chemical nature of the grafted polymer was directly reflected in the water flux-to-rejection ratio and the extent of pore collapse.