Development of a novel 3D culture system for screening features of a complex implantable device for CNS repair

Tubular scaffolds which incorporate a variety of micro- and nanotopographies have a wide application potential in tissue engineering especially for the repair of spinal cord injury (SCI). We aim to produce metabolically active differentiated tissues within such tubes, as it is crucially important to evaluate the biological performance of the three-dimensional (3D) scaffold and optimize the bioprocesses for tissue culture. Because of the complex 3D configuration and the presence of various topographies, it is rarely possible to observe and analyze cells within such scaffolds in situ. Thus, we aim to develop scaled down mini-chambers as simplified in vitro simulation systems, to bridge the gap between two-dimensional (2D) cell cultures on structured substrates and three-dimensional (3D) tissue culture. The mini-chambers were manipulated to systematically simulate and evaluate the influences of gravity, topography, fluid flow, and scaffold dimension on three exemplary cell models that play a role in CNS repair (i.e., cortical astrocytes, fibroblasts, and myelinating cultures) within a tubular scaffold created by rolling up a microstructured membrane. Since we use CNS myelinating cultures, we can confirm that the scaffold does not affect neural cell differentiation. It was found that heterogeneous cell distribution within the tubular constructs was caused by a combination of gravity, fluid flow, topography, and scaffold configuration, while cell survival was influenced by scaffold length, porosity, and thickness. This research demonstrates that the mini-chambers represent a viable, novel, scale down approach for the evaluation of complex 3D scaffolds as well as providing a microbioprocessing strategy for tissue engineering and the potential repair of SCI

Novel hydrogels based on polysaccharides for soft tissue regeneration: preparation and characterization

Předložená diplomová práce se zabývá přípravou, síťováním a fyzikálně-chemickou charakterizací hydrogelů na bázi polysacharidů. Cílem práce bylo vyvinout elastické filmy, které by mohly být použity pro vlhké hojení ran. Teoretická část shrnuje současné způsoby regenerace měkkých tkání a jejích náhradách (ať už se jedná o přírodní nebo syntetické materiály). Zároveň jsou zdůrazněny základní informace o přírodních polysacharidech (chemická struktura, rozpustnost, tepelná a pH stabilita atd.), jejich modifikace a chemické síťování. Experimentální část je zaměřena na modifikaci přírodní gumy Karaya tak, aby transparentní hydrogely měly nastavitelnou hydrolytickou stabilitu. Vzorky byly analyzovány pomocí FTIR, TGA následované vyhodnocením bobtnání a hydrolytické degradace. Z výsledků vyplynulo, že chemická modifikace zvýšila stabilizaci elastického filmu z přírodního polysacharidu ve vodě až na 25 dní. Díky řízené degradaci a vysoké absorpci vody (85 - 96%) jsou tyto nové hydrogely využitelné především pro vlhké hojení ran (např. popálenin).Presented diploma thesis deals with preparation, crosslinking and physico-chemical characterization of natural polysaccharide-based hydrogels. The aim of the work was to evolve elastic thin films with potential application for moist wound healing. The theoretical part summarizes the state-of-art about regeneration of soft tissues and their substitutes (synthetic or nature). There are pointed out the basic information about natural polysaccharide gums (chemical structure, solubility, heat and pH stability etc.), its modification and chemical crosslinking. The experimental part is focused on the modification of natural gum Karaya in order to make transparent hydrogels with adjustable hydrolytical stability. Samples were analyzed by FTIR, TGA followed by evaluation of swelling properties and hydrolytical degradation. Based on the results, chemical modification helped to stabilize polysaccharide hydrogels in water up to 25 days which is useful mainly for moist wound healing (e.g. after burns) because of high values of water uptake (from 85 up to 96%).

Immunogene therapy with fusogenic nanoparticles modulates macrophage response to Staphylococcus aureus.

The incidence of adverse effects and pathogen resistance encountered with small molecule antibiotics is increasing. As such, there is mounting focus on immunogene therapy to augment the immune system's response to infection and accelerate healing. A major obstacle to in vivo gene delivery is that the primary uptake pathway, cellular endocytosis, results in extracellular excretion and lysosomal degradation of genetic material. Here we show a nanosystem that bypasses endocytosis and achieves potent gene knockdown efficacy. Porous silicon nanoparticles containing an outer sheath of homing peptides and fusogenic liposome selectively target macrophages and directly introduce an oligonucleotide payload into the cytosol. Highly effective knockdown of the proinflammatory macrophage marker IRF5 enhances the clearance capability of macrophages and improves survival in a mouse model of Staphyloccocus aureus pneumonia

Nuclear magnetic resonance cryoporometry

Nuclear Magnetic Resonance (NMR) cryoporometry is a technique for non-destructively determining pore size distributions in porous media through the observation of the depressed melting point of a confined liquid. It is suitable for measuring pore diameters in the range 2 nm-1 mu m, depending on the absorbate. Whilst NMR cryoporometry is a perturbative measurement, the results are independent of spin interactions at the pore surface and so can offer direct measurements of pore volume as a function of pore diameter. Pore size distributions obtained with NMR cryoporometry have been shown to compare favourably with those from other methods such as gas adsorption, DSC thermoporosimetry, and SANS. The applications of NMR cryoporometry include studies of silica gels, bones, cements, rocks and many other porous materials. It is also possible to adapt the basic experiment to provide structural resolution in spatially-dependent pore size distributions, or behavioural information about the confined liquid

Advances of nanotechnology in agro-environmental studies

With the increase in the world population and the demand for food, new agricultural practices have been developed to improve food production through the use of more effective pesticides and fertilisers. These technologies can lead to an uncontrolled release of undesired substances into the environment, with the potential to contaminate soil and groundwater. Today, nanotechnology represents a promising approach to improve agricultural production and remediate polluted sites. This paper reviews the recent applications of nanotechnologies in agro-environmental studies with particular attention to the fate of nanomaterials once introduced in water and soil, to the advantages of their use and their possible toxicology. Findings show that the use of nanomaterials can improve the quality of the environment and help detect and remediate polluted sites. Only a small number of nanomaterials demonstrated potential toxic effects. These are discussed in detail

Formulation and evaluation of floating mucoadhesive alginate beads for targetingHelicobacter pylori

Objectives: There are various obstacles in the eradication of Helicobacter.pylori (H. pylori) infections, including low antibiotic levels and poor accessibility of the drug at the site of the infection. This study describes the preparation and characterisation of novel floating-mucoadhesive alginate beads loaded with clarithromycin (CMN) for delivery to the gastric mucosa to improve the eradication of this micro-organism. Methods: Calcium alginate beads were prepared by ionotropic gelation. The formulation was modified through addition of oil and coating with chitosan in order to improve floating, mucoadhesion and modify drug release. Key findings: SEM confirmed the sphericity of the beads with X-ray microtomography (XμMT) showing the 3D structure of the beads with the layered internal structure of the bead and the even distribution of the drug within the bead. This formulation combined two gastro-retentive strategies and these formulations produced excellent in vitro floating, mucoadhesive and drug release characteristics. Enhanced stability of the beads in phosphate buffer raises a potential for the modified formulations to be targeted to regions of higher pH within the gastrointestinal tract with a higher pH. Drug release from these beads was sustained through an unstirred mucin layer simulating in vivo conditions under which the H. pylori resides in the gastric mucosa. Conclusions: This novel formulation will ensure retention for a longer period in the stomach than conventional formulations and control drug release, ensuring high local drug concentrations, leading to improved eradication of the bacteria