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Isolation and characterization of natural active components and their application in food supplements
Šmídová, Veronika ; Němcová, Andrea (referee) ; Skoumalová, Petra (advisor)
The diploma thesis is focused on the characterisation and isolation of naturally occurred active compounds, especially on lipophilic compounds. The next part of the thesis deals with enzymes and their encapsulation into alginate particles to suggest new enzymatic dietary supplement for children with optimal nutrient composition. The theoretical part is focused on the issue of child nutrition, accordingly the necessary nutrients that children need to grow without any health difficulties. It is also focused on the availability of enzymatic food supplements for children. Finally, the theoretical part deals with the lack of chosen natural substances and their effects in connection with cancer. In the experimental part, four types of oils obtained from seeds were characterized. The oils were obtained by two methods: cold pressing with a Yoda kitchen press and extraction in an organic solvent with Soxtherm. Seeds were linseed, sesame, pumpkin, and cumin. With these oils were characterized natural active substances, polyphenols, flavonoids, antioxidants, carotenoids, chlorophylls, and fatty acids. Liposomes were also prepared from selected extracts, in which an effect on intestinal tumour cells were observed. Furthermore, some enzymes were selected, and they were encapsulated into alginate particles with average size 450 µm. In these particles was specified encapsulation efficiency as well as the proteolytic activity after testing in digest juices. The last part of this thesis was focused on the optimization of a complete enzymatic food supplement with addition of alginate particles as enzyme carriers. Samples of these supplements were finally subjected to a sensory analysis.
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Use of microcalorimetry in the study of hydration of biopolymers
Bola, Tomáš ; Smilek, Jiří (referee) ; Krouská, Jitka (advisor)
This master thesis deals with the using of microcalorimetry in the study of hydration of biopolymers. Lactose has been selected together with the other biopolymers although it is not among biopolymers but disaccharides. Selected biopolymers are alginate, dextrane, chitosan and hyaluronan of two molecular weights. Lactose has been selected for these purposes mainly because it is a model example to determine whether or not the reaction to moisture between the other samples and the saturated salt solution occurs. The biopolymer hydration study, as opposed to the commonly used perfusion calorimetry method using the possibility of measuring with adjustable moisture has been used an isothermal microcalorimetry method where at two constant temperatures the reaction of the sample to the different moisture released by the saturated salt solution was monitored.
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Preparation and characterization of hybrid hydrogels matrix
Magera, Lukáš ; Kalina, Michal (referee) ; Smilek, Jiří (advisor)
Submitted bachelor’s thesis aims for preparation and subsequent optimisation of hybrid hydrogels, which represent material with high application potential because of their unique properties. Hybrid hydrogels have extraordinary mechanical resistance and other beneficial properties (e.g. transportation) due to their unique hybrid network structure, strong interpenetrating network entanglement and efficient energy dissipation system. This work engages in chemically-physically crosslinked hybrid gel, namely the combination of alginate-polyacrylamide. The hybrid hydrogel was formed and then underwent the tests of drying and swelling. The main purpose of the experimental part was to determine viscoelastic properties of hybrid hydrogels using standard oscillation tests. Swelling tests showed that the hybrid network containing limitedly swelling alginate gel reaches lower degree of swelling than the single unlimitedly swelling polyacrylamide gel. Viscoelastic tests unveiled an influence on the mechanical properties by value augmentation of the hybrid gel´s storage modulus in comparison with the reference alginate hydrogel. Hybrid hydrogel reported an increase in the linear viscoelastic area compared to the reference polyacrylamide gel, whose hybrid structure prevents from creating microcracks, that caused earlier rupture of the reference polyacrylamide gel.
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Study of barrier and transport properties of polyelectrolytes using diffusion techniques in hydrogels
Valentová, Kristýna ; Kalina, Michal (referee) ; Smilek, Jiří (advisor)
This diploma thesis was focused on study of barrier and transport properties of selected polyelectrolytes in hydrogel matrices by using diffusion techniques. The study of these properties was performed in horizontal diffusion cells where is observed the change in diffusion probe concentration over time. Diffusion experiments were performed on an agarose hydrogel with the addition of alginate, hyaluronic acid, polystyrene sulfonate, humic acids and as a model probe rhodamine 6G was used. Important parts of this thesis are also the methods which characterize the substances and hydrogel matrices such as rheology and potentiometric titration. The main aim of this diploma thesis was to investigate the effect of interactions between passing model dye (rhodamine 6G) and the appropriate gel (agarose + polyelectrolyte) on the fundamental diffusion parameters (effective diffusion coefficient, lag time, etc.).
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Adhesion of (Hydro)gel Materials to the Surface – Physico-chemical Description and Biological Concequences
Žibeková, Lucia ; Mravec, Filip (referee) ; Kalina, Michal (advisor)
This Bachelor´s thesis deals with study of adhesion of hydrogels materials to the surface. Three biopolymers were selected for this purpose, one of which is psychically cross-linked (agarose) and two are chemically cross-linked (based on sodium alginate cross-linked with Ca2+ and polyvinyl alcohol cross-linked with borax). In the experimental part of the work, adhesion was first examined by a visual method on an inclined plane. The tilt was at different angles (30°, 45°, 90°) where it was observed whether the hydrogel was able to attach to a solid surface or slip down. The agarose was examined in different concentrations from 0.5 wt% to 4 wt% in both methods. Also, the alginate gel was examined at various concentrations with two different cross-linked agent. However, alginate was only used in the visual method because of its inhomogeneity. In contrast, the PVA gel was examined at 10:1 and 4:1 ratios for the both methods. The second method in the experimental part was the tensile test carried out on the Inova apparatus – hydraulic pulsator, and thus the methodology for the study of physical and chemically cross-linked hydrogels was optimized. With increasing concentration of agarose gels, adhesion strength and work increased as well. For PVA gels, the adhesion work was higher, but the adhesive strength remained approximately the same as for the agarose gels.
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Preparation and characterization of enzyme-containing wound dressings
Jurová, Bronislava ; Szotkowski, Martin (referee) ; Skoumalová, Petra (advisor)
This diploma thesis is focused on the study and characterization of nanofiber wound dressings from natural biopolymers. Proteolytic enzymes, specifically collagenase and bromelain, were incorporated into these nanofiber covers. The theoretical part deals with a brief description of skin wounds, their healing and skin covers. There are also methods for preparing these covers and the methods used to characterize them. The practical part deals with the optimization of the preparation of nanofiber coatings based on polyhydroxybutyrate, gelatin, alginate and chitosan. These nanofibers were then enriched with active substances and their gradual release into the model environment was monitored. Finally, their proteolytic activity was determined for these substances.
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Incorporation of microbial cells in hydrogel carriers
Orišková, Sofia ; Pekař, Miloslav (referee) ; Sedláček, Petr (advisor)
The presented diploma thesis focuses on the use of plant growth promoting bacteria as an ecological alternative to conventional fertilizers. The incorporation of bacterial cells into hydrogel carriers is already a well-studied topic, but due to its disadvantages it has not yet found wider application in agriculture. This work offers a novel concept of encapsulating bacteria by gelation directly from the culture. This is achieved by crosslinking the bacterial alginate produced by the model microorganism Azotobacter vinelandii. Since this process was not described before, first its optimization was needed. Alginate production was determined gravimetrically, and its parameters were further characterized using available analytical methods – infrared spectroscopy to monitor structural parameters (monomer composition and the extent of acetylation), dynamic light scattering to characterize the size distribution and AF4-MALS-dRI to obtain the molecular weight. Bacterial PHB production was also investigated using gas chromatography and infrared spectroscopy. The second part of the work is focused on the optimization of the gelling process using bacterial alginate from the culture and CaCl2 as a crosslinking agent. Rheological experiments were used as a tool in understanding the viscoelastic properties of the prepared gels. Gelation was demonstrated within the first day after inoculation. Maximum production of alginate (1,9 ± 0,3) g/l was reached on the fourth day after inoculation. It was found that the addition of 5 g/l of calcium carbonate promotes the production of alginate. Nevertheless, further addition of CaCO3 (30 g/l) showed adverse effects on the molecular weight and is therefore not recommended. Production of PHB was confirmed by both FTIR and GC measurements, with a maximum yield of (23 ± 3) % CDW. Rheological testing confirmed that the product of the crosslinking was a gel. It was found that the crosslinker concentration plays an important role at time 0 min of the gelation, forming a denser network in the structure and causing higher rigidity. Using the highest studied concentration of CaCl2, the critical strain reached values of (5,0 ± 0,7) %. Finally, the incorporation of bacterial cells into the hydrogel was confirmed using fluorescence microscope.
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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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New possibilities in wound healing
Nováková, Laura ; Szotkowski, Martin (referee) ; Skoumalová, Petra (advisor)
The diploma thesis is focused on the study of fibrous wound dressings prepared by electrospinning method from natural biopolymers. Three active ingredients were added to the dressings: ampicillin, ibuprofen and collagenase, which are responsible for relieving pain, reducing the risk of infection and selectively removing necrotic tissue in the wound. The theoretical part describes the therapeutic dressings currently available on the market and the most common methods of nanofiber production. The experimental part evaluates the optimization of the preparation of gelatin, alginate and chitosan fibrous wound dressings, which were subsequently enriched with active substances and their gradual release into the model environment was determined spectrophotometrically. Antimicrobial effects against E.coli and S. epidermidis strains andantifungal activity against C. glabrata yeast were monitored. Finally, two cytotoxicity tests on the human keratinocyte cell line HaCaT confirmed the safety of the prepared products, which can serve as bioactive skin dressings in the future.
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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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