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Preparation of microfiber structures based on PHB copolymer
Kecíková, Alžbeta ; Brtníková, Jana (oponent) ; Přikryl, Radek (vedoucí práce)
Master’s thesis deals with the preparation of microfibrous structures based on poly(3-hydroxybutyrate) (P3HB) and its copolymer with poly(4-hydroxybutyrate) using centrifugal spinning technology. The microfibers were modified by the addition of oligomer P3HB and a plasticizer. The centrifugal spinning process was optimized for each material by solution viscosity, polymer molecular weight, speed of spineret, and presence of solvent. One of the part of optimalization was the addition of formic acid and acetic acid to the dissolution system. Microfibrous structures have been investigated in terms of morphology, mechanical properties, wetting and biocompatibility. P3HB fibers were also surface treated with lipase solutions to reduce their hydrophobicity. The prepared bulky fiber cocoons have a suitable 3D microstructure for monitoring and testing biological properties in vitro. Thus, a potential application of microfiber structures is as 3D cell culture carriers in an scaffolds in vitro system.
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Optimization of methods for protein analysis released from thermosensitive hydrogel
Lysáková, Klára ; Brtníková, Jana (oponent) ; Vojtová, Lucy (vedoucí práce)
Proposed diploma thesis is focused on the release evaluation methods of both model proteins (albumin and lysozyme) and tissue healing protein (stable fibroblast growth factor 2; FGF2-stab) from the “smart” hydrogel carrier based on the biodegradable thermosensitive PLGA-PEG-PLGA triblock copolymer. In the theoretical part, a brief overview of thermosensitive polymers, their properties, structure and utilization of aforementioned proteins is described. Moreover, types of interactions that may occur between proteins and the polymeric carrier including drug release models are mentioned. In the experimental part, the structure of PLGA-PEG-PLGA triblock copolymer was characterized by the gel permeation chromatography and 1H NMR while its visco-elastic properties including sol-gel transition were evaluated by the rheological analysis. Thesis goal was targeted to better understand the release of newly-patented FGF2-stab protein from injectable PLGA-PEG-PLGA hydrogel formed at physiological temperature. The amount of released FGF2-stab was measured by the UV-VIS spectrophotometer in the presence of a Bradford reagent that binds proteins resulting in shifting their absorption maxima from 280 nm to 595 nm. For the comparison, SDS-page electrophoresis, dividing protein by molecular weight, has been used. It has been find out, that model proteins, which were in different size but predominantly non-polar on their surface exhibited two-stage release dependent on both the diffusion and polymer degradation, whereas, FGF2-stab (25 kg.mol-1) showed controlled one-stage first-order release from the PLGA-PEG-PLGA hydrogel forced only by diffusion, since it is predominantly a polar molecule located probably at the hydrogel micelle surface and not in their core. These results are very important to tune the protein release from hydrogel carriers to meet their certain application, in this case specifically in controlled tissue regeneration.
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Novel Antibacterial Collagen Scaffolds for Regenerative Medicine
Dorazilová, Jana ; Brtníková, Jana (oponent) ; Vojtová, Lucy (vedoucí práce)
This master’s thesis deals with the characterisation of 3D porous collagenous sponges enriched with selected antibacterial agents. The literature part of the thesis focuses on the overview of biomaterials and biopolymers with the emphasis on collagen and chitosan, outlines the antibacterial properties of nanoparticles and reviews current aspects of using selenium nanoparticles as an antibacterial agent. For the purpose of this work, two types of antibacterial additives were used – biopolymeric chitosan and selenium nanoparticles. Preparation of 3D porous structure was achieved using the freeze drying method. Mechanical properties of prepared biopolymeric matrices were improved by chemical crosslinking in the presence of carbodiimide. Predominantly physiochemical methods were used for characterization of prepared collagenous sponges. For microstructure analysis, pore size determination, visualisation of nanoparticles and their distribution inside the porous structure, scanning electron microscopy (SEM) with energy dispersive x ray optical analysis (EDX) was used. Parameters such as total porosity, swelling ratio, weight loss during degradation in water and enzymatic environment were evaluated by suitable gravimetric methods. Fourier transformed infrared spectroscopy with attenuated total reflectance (ATR-FTIR) was used to determine changes in the chemical structure of collagenous matrices before and after addition of the antibacterial agents. Percentage release of nanoparticles was evaluated using optical emission spectroscopy with inductively coupled plasma (ICP OES). Evaluation of antibacterial properties of tested samples was carried out mainly by the agar diffusion disk method and the macrodilution broth method. In the conducted research we were able to determine the influence of selected antibacterial additives on the physiochemical properties of 3D collagenous matrices. Their antibacterial activities showed a positive effect on bacterial inhibition of both chitosan and selenium nanoparticles with respect to their concentrations. The designed materials could be further utilized for bio medicinal applications, especially in the field of soft tissue regeneration.
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Effect of bioceramic additives on morphology, physical and biological properties of collagen scaffolds for bone tissue engineering
Klieštiková, Nikola ; Poláček, Petr (oponent) ; Brtníková, Jana (vedoucí práce)
The diploma thesis deals with preparation of three-dimensional porous collagen composite scaffolds for bone tissue engineering and study of the effect of addition of bioceramic particles on morphological, biomechanical and biological properties. Theoretical part describes biomaterials and bioceramic particles used for scaffolds in bone tissue engineering and their fabrications method. As for experimental part, samples were prepared by the freeze-drying method. As tested material, type I collagen from porcine and bovine sources was combined with hydroxyapatite and mixture of -tricalcium phosphate and -tricalcium phosphate in ratios 1 : 1, 1 : 2 and 2 : 1. The effect of bioceramics solubility and particle sizes on scaffolds morphology, biomechanics and biocompatibility was evaluated. Addition of bioceramic particles changed the morphology of the samples. The pore size decreased, whereas the porosity was nearly the same in all tested samples. Bioceramic particles also made the collagen matrix of the scaffolds less hydrophilic, moreover they stabilized the scaffolds against the effect of enzymatic degradation. The biomechanical properties of the samples were tested in both dry and hydrated state. In dry state, the pure bovine collagen scaffolds reached the highest compressive strength, contrary in hydrated state, the samples containing bioceramic particles reached the highest value. None of the samples was cytotoxic and the most preferable environment for cell adhesion and proliferation was in the pure bovine collagen scaffolds and also in the composite scaffolds with ratio HAp : -TCP : 1 : 1.
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Průzkum čtenářské úrovně žáků 2. ročníků základních škol
BRTNÍKOVÁ, Jana
Diplomová práce s názvem Průzkum čtenářské úrovně žáků 2. ročníků na ZŠ se zaměřuje na výskyt nejčastějších čtenářských nedostatků v počátečním čtení. Práce je rozdělena na část teoretickou a výzkumnou. Teoretická část se zabývá čtením z hlediska vzdělávacího systému, seznamuje nás s fyziologickými a psychologickými pochody při čtení, zmiňuje historické i současné metody výuky čtení, základní druhy a kvality čtení, etapy rozvoje čtenářských dovedností, čtenářské nedostatky, nesprávné čtecí návyky a popisuje práci se zaostávajícími čtenáři. Výzkumná část je věnována výzkumu, jeho realizaci a vyhodnocení. Hlavním cílem je zjistit, co dělá začínajícím čtenářům největší problémy a jaká je celková úroveň čtenářských dovedností žáků 2. ročníků. Výzkumná část převážně interpretuje výsledky nahrávání čtenářského výkonu.
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