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Phospholipids as the basis of biodegradable delivery systems
Burdíková, Jana ; Čeppan, Michal (referee) ; Mravec, Filip (advisor)
This thesis is focused on investigation of phospholipid-hyaluronan system. First, appropriate method for preparation of bulk solution of phospholipid/lipid and suitable fluorescence probe were chosen. Sonification was selected as a method for preparation of bulk solution and pyrene was chosen as a fluorescence probe. From the group of phospholipids lecithin was selected. Next to phospholipid, lipid with no phosphate group (DPTAP) was utilized for comparison, alternatively a mixture of lipid (DPTAP) and phospholipid (DPPC). Instead of hyaluronan another polyelectrolytes (sodium polystyrene sulfonate, sodium alginate) were used too. Measurements were performed in water environment and in phosphate buffer saline (PBS). All investigation was accomplished by fluorescence spectroscopy and dynamic light scattering.
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The Utilization of Light Scattering Techniques for the Study on Hydrogel Gelation
Candráková, Simona ; Smilek, Jiří (referee) ; Kalina, Michal (advisor)
This bachelor thesis focuses on the study of hydrogel gelation using light scattering methods. For these purposes two different biopolymers (agarose and sodium alginate), with different sol to gelation phase transition, were selected. In the case of agarose, the gelation is caused by temperature change. On the other side, the gelation of sodium alginate is initiated by addition of polyvalent cations. In the experimental part of the work, agarose gelation was studied by DLS, temperature gradients of agarose solutions (40 – 30 °C) were measured. During the measurement the particle size distributions in the solutions having different concentrations were monitored as well as the temperature influence on the diffusion coefficient. The DLS method provided the values of the gelling temperatures of the solutions at a concentration of 0,5 wt.% and 1,0 wt.%. Furthermore, the DLS microrheology method was used, where temperature dependences of viscoelastic characteristics of agarose solutions (40 – 30 °C) were also measured, from which the gelling temperatures of agarose solutions of 0,1 wt.% and 0,5 wt.% were evaluated. The classical rheology was also used in the work to compare the viscoelastic behaviour of the samples and to determine the gelation point of the agarose solutions. By this method were determined gelling temperatures for all studied concentration of agarose (0,1 wt.%, 0,5 wt.% and 1,0 wt.%). The DLS method was also used to monitor the interactions of sodium alginate with the addition of Ca2+ ions, these interactions were then also evaluated and discussed in the experimental part.
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Mechanical and Transport Properties of Hybrid Hydrogel Systems
Klímová, Eliška ; Sedláček, Petr (referee) ; Kalina, Michal (advisor)
This master´s thesis deals with the study on mechanical and transport properties of hybrid hydrogel systems. Considering applications of hydrogels, especially in chemical industry, pharmacy, or eventually medical applications, for the study gellan and alginate-based hydrogels were selected. In order to compare individual characteristics physical and hybrid hydrogels were prepared. Gellan hydrogels were prepared in deionization aqua solution, calcium chloride dihydrate and tween 80 solution. Alginate hydrogels were prepared in calcium chloride dihydrate solution as well, and polyacrylamide with N,N´–methylenbisacrylamide. For the study of mechanical properties moisture analyser and rheology measurements were selected. Transport properties were studied using the diffusion experiments combined with UV-VIS spectroscopic detection. Concluding of this thesis is summarization of measured values, which provides comprehensive review of the problematics. It was discovered that the conveniently selected concentrations of structural components of hydrogel matrix and the additives can influence both the mechanical as well as the transport properties of studied hydrogels.
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Encapsulation of selected natural extract for food application.
Vyskočilová, Terezie ; Kočí, Radka (referee) ; Márová, Ivana (advisor)
This diploma thesis deals with encapsulation of natural extracts. In the theoretical part the methods of encapsulation, materials for particle preparation, as well as application of encapsulation techniques in food industry were described. In experimental part selected natural extracts of propolis, green barley and probiotics were characterized. There substances were encapsulated into alginate and chitosan. In the total of 25 types of prepared particles long-term stability in some model physiological conditions as well as in four different model foods was evaluated. Additionally, stability of selected particles in several real milk-based products was followed too. The stability of particles was determined spectroptohometrically. In natural extract a content of polyphenols, proteins, chlorophylls, as well as total antioxidant activity were analysed. To analysis of probiotics optical and fluorescence microscopy were used. In propolis and green barley antimicrobial activity was tested too. Moreover, in the sample of propolis also cytotoxic assay was applied. Agar-chitosan was chosen as the best shell material for propolis due to its optimal stability in model physiological conditions as well as model foods. Liposomes were evaluated as unstable and were not recommended for further application. As the suitable shell material for powdered green barley starch-alginate (rate 1:4) and agar-chitosan were proposed, while the second one showed better stability for released proteins. Agar-chitosan shell material was usable for fresh green barley too. For probiotics encapsulation alginate or alginate-starch were chosen because of their porosity and possibility of nutrients diffusion. In real foods the best results were reached with application of probiotic particles into milk. Coencapsulation of powdered barley and probiotics did not confirm inhibition of culture growth. Neither the antimicrobial effect of propolis and barley nor the cytotoxic effect of propolis were confirmed.
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Encapsulation of active substances into nanofibers and possibilities of their application
Procházková, Lucie ; Pernicová, Iva (referee) ; Skoumalová, Petra (advisor)
The master thesis was based on the optimization of the production of nanofiber covers and to gaine the product for subsequent functional use. The production of nanofiber covers was made by electrospinning and forcespinning from selected materials. Polyhydroxybutyrate, gelatin, chitosan and alginate were used as starting materials. After successful optimization, these materials were enriched with active ingredients ampicillin and ibuprofen for the functionalized use of covers for more effective wound healing. The theoretical part was focused on the issue of skin, healing processes, types of wounds and nanofibers, the characterization of selected starting materials for the formation of nanofibers was also mentioned. The practical part was based on the lengthy optimization of the preparation of fiber covers and later enriched with active ingredients. Furthermore, combined covers made of different materials with contents of both active ingredients were designed. This was followed by the characterization of all prepared covers from the point of view of stability in the short and long term. The gradual release of active ingredients was determined spectrophotometrically and by hifh performance liquid chromatography. It was also important to determine the antimicrobial activity of selected active substances. At the end of all testing, combined coatings containing both active ingredients were used for safety testing with human keratinocyte cells (HaCaT). Safety testing was based on determining the viability of human cells using the MTT test, to verify the LDH test. A scratch test was also performed, a wound healing test after the application of devised combined covers.
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Development and characterization of hydrogel carriers for modern agricultural applications
Mai, Thuy Ha ; Kalina, Michal (referee) ; Sedláček, Petr (advisor)
The bachelor thesis is focused on characterization of hydrogel carriers and their application in agriculture. Entrapment of active phase within the hydrogel matrix would allow release of its content in controlled rate. The theoretical part describes encapsulation techniques and suitable biocompatible materials involved in encapsulation. For the experimental part, the release of soil bacteria Cupriavidus necator H16 from alginate matrix was examined. The focus was on the release rate of bacteria via diffusion and later, on the release rate of bacteria encouraged by chemical or enzymatic reagents. Further modification of carrier was suggested by adding hydrophobic barrier composed of polyhydroxybutyrate (PHB). The modified alginate beads seemed to be more resistant toward enzymatic or chemical agents, thus the process of the release was to some extent suppressed. This fact might suggest PHB as a possible vessel for optimization of controlled release system of hydrogel carriers.
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Hydrogel carriers of microorganisms for modern environmental aplications.
Súkeník, Martin ; Slaninová, Eva (referee) ; Sedláček, Petr (advisor)
This bachelor thesis deals with the optimization of cultivation and gelation procedures by bacterium Azotobacter vinelandii, which belongs to a PGPR group, capable of synthesis of intracellular polyhydroxyalkanoates and extracellular alginate. The alginate produced by the bacteria is crosslinked with a calcium chloride solution to form a hydrogel carrier containing the described bacterium. This unconventional concept simplifies bacterial encapsulation and production of hydrogel carriers, furthermore expands its usability in modern environmental applications. In the first part of this thesis, three strains (DSM 85, 87, 720) were selected to measure a concentration of produced alginate, its molecular weight was determined by a SEC-MALS technique and M/G ratio was analysed by infrared spectroscopy. The Azotobacter vinelandii strain DSM 87 reached the highest concentration of the alginate (4.9 ± 0.6) g/l by gravimetric determination on the 5th day of cultivation. The concentration of produced PHB ranges from 48 to 6 % of CDW, which was determined by gas chromatography. Nevertheless, the strain DSM 720 showed the best gelation properties and was used in the second part of this thesis for further optimization of the cultivation process, and for the analysis the formation of gelation procedure under different temperatures, gelation time and concentrations of crosslinking solution. The formed gels were compared by rheological measurements of amplitude and frequency tests.
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Dry gel forms - aerogels, xerogels - preparation, properties and potential use.
Sedlář, Marian ; Smilek, Jiří (referee) ; Sedláček, Petr (advisor)
The content of this bachelor thesis was to find out what influence has the drying of hydrogel on its internal structure on an appropriate model hydrogel system. In the theoretical part of this thesis selected methods of structural analysis that are commonly used for characterization on gels in their original hydrated state and in dry state were described. In the practical part, agarose based hydrogels with different concentrations and with various additions of polyelectrolytes were chosen as the model hydrogel system. Cryo-scanning electron microscopy and turbidimetry were used for the structural analysis of hydrogels. For the dry gels, scanning microscopy and mercury porosimetry were used. The obtained data and results for our model system can be use in analyse of other hydrogel systems and their dry forms.
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Encapsulation of soil bacteria in hydrogel carriers
Orišková, Sofia ; Smilek, Jiří (referee) ; Sedláček, Petr (advisor)
The goal of this bachelor thesis was both to review existing literature regarding the topic of inoculation of soil bacteria and test a relevant encapsulation method and optimize it. The evaluation process involved the study of various encapsulation techniques that involve hydrogel carriers suitable for agronomic purposes. Encapsulation allows controlled release of soil bacteria, and protects and stabilizes it, while ensuring longer shelf life. For the practical testing, Cupriavidus necator H16 was chosen as a nitrogen fixing bacteria for the inoculation. Through an ionic gelation method, it was encapsulated into alginate carriers, forming matrix microcapsules. Sodium alginate reacts with the cross-linking agent calcium chloride to form the capsules. The impact on the quality of the product was tested through several variables. What was revealed was that molecular weight of alginate was proven to have a significant impact. Alginate with higher molecular weight was shown to be suitable for the given system. The most desirable environment was distilled water or a TRIS-HCl buffer. However, the TRIS-HCl was unsuitable in cases of retrieving a dry product by lyophilization. Eventually, the viability of encapsulated cells was examined, and it was proven that encapsulated bacteria endure the process of encapsulation in the above-mentioned environment and they were gradually released from the carrier.
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Preparation of bioactive wound dressings and testing their interaction with human cells
Bendová, Agáta ; Márová, Ivana (referee) ; Skoumalová, Petra (advisor)
The thesis was focused on the preparation and optimization of the preparation of wound dressing from materials with bioactive ingredients. In this work were prepared nanofiber dressings based on polyhydroxybutyrate and non-fibrous dressings from alginate and chitosan. Nanofibers were prepared by electrospinning and forcespinning methods. The bioactive components, which were used to functionalize the prepared dressings, were plant extracts, clotrimazole, ampicillin, lysozyme, and proteolytic enzymes. The theoretical part is focused on the description of the use of nanofibrous and non-fibrous materials in medicine, characterization of materials for the production of wound dressings and bioactive components. Furthermore, this section describes the methods used to prepare and characterize wound dressings. In the practical part were prepared aqueous and oil extracts from selected plants. Extracts were characterized for polyphenols content and antioxidant activity. PHB-based nanofibers were prepared using electrospinning and forcepinning methods. Nanofibers were enriched with selected plant oil extracts and clotrimazol. Modified nanofibres were detemined for antioxidant activity, short-term and long-term stability. Non-fibrous wound dressings were prepared from alginate and chitosan. These dressings were functionalized by the addition of selected aqueous extracts, ampicillin, lysozyme, papain, bromelain, and collagenase. Non-fibrous wound dressings were determined for antioxidant activity, short-term stability and proteolytic activity. The prepared wound dressings were tested for their antimicrobial effects on cultures of Micrococcus luteus, Serratia marcescens, Staphylococcus epidermidis and Escherichia coli. In conclusion, successfully prepared bioactive wound dressings with antioxidant and antimicrobial agents were tested for safety on human cells. The determination was performed using the MTT cytotoxicity test on human keratinocytes.
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