Evaluation of ethanol-induced chitosan aerogels with human osteoblast cells.

The following article provides an insight into the production of chitosan aerogels as potential materials for tissue engineering. Chitosan aerogels were prepared following two different protocols: formation in ethanol and formation in sodium hydroxide in an ethanol solution. The main objective was to apply a new route to obtain chitosan aerogels with no external cross-linkers and compare the mentioned preparation approaches. Forming chitosan aerogels in ethanol implies a simple, environmentally friendly, and efficient method. The prepared materials showed specific surface areas of up to 450 m2/g, highly porous networks and great mechanical properties. In vitro degradation studies revealed high stability for up to 10 weeks. The differences between the samples were significant. While the chitosan aerogels prepared in ethanol showed superior textural, morphological and mechanical properties, the chitosan aerogels prepared in the sodium hydroxide solution proved that a considerable influence on end properties could be made simply by adjusting the ageing medium. In vitro cell analysis with primary human osteoblasts showed good biocompatibility and pointed towards the potential use of these aerogels for orthopedic applications. This testing showed further that adjustments in structural properties by sodium hydroxide also come with a cost regarding their suitability to host bone cells.

A Brief Evaluation of Pore Structure Determination for Bioaerogels.

This review discusses the most commonly employed methods for determining pore size and pore size distribution in bioaerogels. Aerogels are materials with high porosity and large surface areas. Most of their pores are in the range of mesopores, between 2 and 50 nm. They often have smaller or larger pores, which presents a significant challenge in determining the exact mean pore size and pore size distribution in such materials. The precision and actual value of the pore size are of considerable importance since pore size and pore size distribution are among the main properties of aerogels and are often directly connected with the final application of those materials. However, many recently published papers discuss or present pore size as one of the essential achievements despite the misinterpretation or the wrong assignments of pore size determination. This review will help future research and publications evaluate the pore size of aerogels more precisely and discuss it correctly. The study covers methods such as gas adsorption, from which BJH and DFT models are often used, SEM, mercury porosimetry, and thermoporometry. The methods are described, and the results obtained are discussed. The following paper shows that there is still no precise method for determining pore size distribution or mean pore size in aerogels until now. Knowing that, it is expected that this field will evolve in the future.

Influence of the Impregnation Technique on the Release of Esomeprazole from Various Bioaerogels.

The presented study shows the possibility of using bioaerogels, namely neat alginate, pectin, chitosan aerogels, and alginate and pectin aerogels coated with chitosan, as drug delivery systems for esomeprazole. Two different techniques were used for the impregnation of esomeprazole: Supercritical impregnation, and diffusion via ethanol during the sol-gel synthesis. The prepared samples were characterized by employing N2 adsorption-desorption analysis, TGA/DSC, and FTIR. The achieved loadings were satisfactory for all the tested samples and showed to be dependent on the technique used for impregnation. In all cases, higher loadings were achieved when impregnation via diffusion from ethanol was used. Extensive release studies were performed for all impregnated samples. The in vitro dissolution profiles were found to be dependent on the carrier and impregnation method used. Most importantly, in all cases more controlled and delayed release was achieved with the bioaerogels compared to using pure esomeprazole.

Platelets express adaptor proteins of the extrinsic apoptosis pathway and can activate caspase-8.

BACKGROUND: Apoptotic pathways in platelets are important for their survival and function. Platelet apoptosis may be involved in the pathogenesis of immune thrombocytopenia (ITP), an autoimmune-mediated disease. In contrast to the intrinsic apoptosis pathway, not much is known about the extrinsic pathway mechanisms in platelets. OBJECTIVES: To investigate the expression of proteins involved in the extrinsic apoptosis pathway, including the death receptors, adaptor and regulator proteins in human platelets. To determine a possible trigger of the extrinsic apoptosis pathway in platelets. METHODS: To investigate the expression of key markers of the extrinsic pathway we used targeted immunofluorescence and flow cytometry assays. To study their expression and interaction we performed Western blotting and co-immunoprecipitation. Treated platelets with different apoptosis triggers were subjected to flow cytometry. RESULTS: We could identify the protein expression of the pro-apoptotic proteins TRADD (Tumor Necrosis Factor Receptor type 1- Associated DEATH Domain protein), TRAF2/5, (TNF Associated Factor) and DEDAF (Death Effector Domain- Associated Factor), FADD (Fas-Associated protein with death domain) as well as the anti-apoptotic proteins DJ-1 (Deglycase 1) and c-FLIP in human platelets. ABT-737 treatment induced a disruption in the co-localization of DJ-1 with FADD. Platelets treated with ABT-737 showed an activation in caspase-3 and -8. The exposure to TNF (Tumor Necrosis Factor), FasL (Fas ligand), and TWEAK or to plasma derived from ITP patients, did not lead to changes in caspase-3 and -8 activation in platelets. CONCLUSIONS: Human platelets express some proteins of the extrinsic apoptosis pathway which can be modulated only by ABT-737 treatment. However so far, no other apoptosis trigger or interaction with an external receptor have been yet identified.

Optimizing the α1B-adrenergic receptor for solution NMR studies.

Sample preparation for NMR studies of G protein-coupled receptors faces special requirements: Proteins need to be stable for prolonged measurements at elevated temperatures, they should ideally be uniformly labeled with the stable isotopes 13C, 15N, and all carbon-bound protons should be replaced by deuterons. In addition, certain NMR experiments require protonated methyl groups in the presence of a perdeuterated background. All these requirements are most easily satisfied when using Escherichia coli as the expression host. Here we describe a workflow, starting from a temperature-stabilized mutant of the α1B-adrenergic receptor, obtained using the CHESS methodology, into an even more stable species, in which flexible parts from termini were removed and the intracellular loop 3 (ICL3) was stabilized against proteolytic cleavage. The yield after purification corresponds to 1-2 mg/L of D2O culture. The final purification step is ligand-affinity chromatography to ensure that only well-folded ligand-binding protein is isolated. Proper selection of detergent has a remarkable influence on the quality of NMR spectra. All optimization steps of sequence and detergent are monitored on a small scale by monitoring the melting temperature and long-term thermal stability to allow for screening of many conditions. The stabilized mutant of the α1B-adrenergic receptor was additionally incorporated in nanodiscs, but displayed slightly inferior spectra compared to a sample in detergent micelles. Finally, both [15N,1H]- as well as [13C,1H]-HSQC spectra are shown highlighting the high quality of the final NMR sample. Importantly, the quality of [13C,1H]-HSQC spectra indicates that the so prepared receptor could be used for studying side-chain dynamics.

Preparation and Characterization of Chitosan-Coated Pectin Aerogels: Curcumin Case Study.

The following study describes the preparation of pectin aerogels and pectin aerogels coated with an external layer of chitosan. For the preparation of chitosan-coated pectin aerogels, a modified coating procedure was employed. Since pectin as well as pectin aerogels are highly water soluble, a function of chitosan coating is to slow down the dissolution of pectin and consequently the release of the active substances. Textural properties, surface morphologies, thermal properties, and functional groups of prepared aerogels were determined. Results indicated that the coating procedure affected the textural properties of pectin aerogels, resulting in smaller specific surface areas of 276 m2/g, compared to 441 m2/g. However, chitosan-coated pectin aerogels still retained favorable properties for carriers of active substances. The case study for prepared aerogels was conducted with curcumin. Prior to in-vitro release studies, swelling studies were performed. Curcumin's dissolution from both aerogels showed to be successful. Pectin aerogels released curcumin in 3 h showing a burst release profile. Chitosan-coated pectin aerogels prolonged curcumin release up to 24 h, thus showing a controlled release profile.

Preparation and characterization of polysaccharide - silica hybrid aerogels.

Hybrid aerogels based on polysaccharides - silica were prepared and characterized. Tetramethylorthosilicate (TMOS) was used as inorganic precursor and various polysaccharides (alginate, pectin, xanthan and guar) were used as organic precursors. TMOS was added to polysaccharide aqueous solutions, resulting in stable wet gels. There were no additional chemicals or cross-linkers in the process. Produced wet gels were dried under supercritical conditions with CO2 in order to preserve their structure. The nitrogen adsorption results were compared to pure polysaccharide aerogels, prepared in our previous research. It is shown, that the addition of silica to pectin, xanthan, alginate and guar significantly improved their structural properties, primarily seen in the drastic increase of the surface area. Guar-silica aerogels reached the highest surface area of 679 m2 g-1. The thermal properties, including thermal degradation and thermal conductivity were highly improved. Among the prepared hybrid aerogels, pectin-silica samples had the lowest thermal conductivity of 19 mWm-1 K-1.

The anxiolytic effects of atorvastatin and simvastatin on dietary-induced increase in homocysteine levels in rats.

The aim of this study was to evaluate the effects of atorvastatin and simvastatin on behavioral manifestations that followed hyperhomocysteinemia induced by special dietary protocols enriched in methionine and deficient in B vitamins (B6, B9, B12) by means of alterations in anxiety levels in rats. Simultaneously, we investigated the alterations of oxidative stress markers in rat hippocampus induced by applied dietary protocols. Furthermore, considering the well-known antioxidant properties of statins, we attempted to assess their impact on major markers of oxidative stress and their possible beneficial role on anxiety-like behavior effect in rats. The 4-week-old male Wistar albino rats were divided (eight per group) according to basic dietary protocols: standard chow, methionine-enriched, and methionine-enriched vitamins B (B6, B9, B12) deficient. Each dietary protocol (30 days) included groups with atorvastatin (3 mg/kg/day i.p.) and simvastatin (5 mg/kg/day i.p.). The behavioral testing was performed in the open field and elevated plus maze tests. Parameters of oxidative stress (index of lipid peroxidation, superoxide dismutase, catalase activity, glutathione) were determined in hippocampal tissue samples following decapitation after anesthesia. Methionine-load dietary protocols induced increased oxidative stress in rat hippocampus, which was accompanied by anxiogenic behavioral manifestations. The methionine-enriched diet with restricted vitamins B intake induced more pronounced anxiogenic effect, as well as increased oxidative stress compared to the methionine-load diet with normal vitamins B content. Simultaneous administration of statins showed beneficial effects by means of both decreased parameters of oxidative stress and attenuation of anxiety. The results obtained with simvastatin were more convincible compared to atorvastatin.