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Hyaluronan hydrogels for medical applications
Janča, David ; Krouská, Jitka (referee) ; Venerová, Tereza (advisor)
This work studies properties of hydrogels prepared by the interaction of polyelectrolyte hyaluronate with cationic surfactant cetyltrimethylammonium bromide (CTAB) in a model physiological solution of 0,15 M NaCl. Effects of different pH and final pH of different samples of hydrogels were studied. Furthermore, stability of hydrogels was observed by fluorescence spectroscopy, where pyrene was used as a fluorescence probe, and isothermal microcalorimetry. It was found that low temperature disrupts gel integrity and it is not recommended to expose hydrogels to other than a neutral pH.
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Study of the interaction of hyaluronan and biocompatible amphiphilic substances
Cigánková, Hana ; Krouská, Jitka (referee) ; Mravec, Filip (advisor)
This thesis focuses on the study of the interactions between hyaluronan and selected phospholipids and their mixtures in water or buffer. First, the aggregation behavior of these lipids was measured, afterwards the influence of hyaluronan was studied. This behavior was investigated by fluorescence spectoscopy using pyrene as fluorescence probe. The Critical Agregation Concentration of the lipids before and after addition of hyaluronan was determined. The measurements revealed that the addition of hyaluronan has an observable influence on the aggregate behavior of the selected lipids and their mixtures.
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Fluorescence probes in investigation of associative colloid domains.
Mischingerová, Monika ; Krouská, Jitka (referee) ; Mravec, Filip (advisor)
The aim of this thesis was to determine how the concentration of the fluorescent probe (pyrene) affects the critical micelle concentration. In this thesis were used all types of surfactants (anionic, cationic and nonionic). As representatives of the anionic surfactants were chosen sodium dodecylsulfate (SDS) and sodium dodecylbenzenesulfate (SDBS), as the cationic surfactant was chosen cetyltrimethylamonium bromide (CTAB) and as the nonionic polyoxyethylene(9.5)octylphenol (Triton X–100). The critical micelle concentration of these surfactants were measured with a pyrene at three different concentrations. All data obtained by fluorescence spectroscopy were fitted by a Boltzmann-type sigmoid. It was found out that in the case of the anionic surfactants there is some correlation between the critical micelle concentration of the surfactant and the concentration of the fluorescent probe. In both cases (SDS, SDBS) the value of the critical micelle concentration increased with increasing concentration of the fluorescent probe. For the other surfactants (cationic and nonionic) was observed no change in the critical micelle concentration values depending on the concentration of pyrene.
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Aggregation processes in amphiphil-polymer system
Šejnohová, Michaela ; Márová, Ivana (referee) ; Pekař, Miloslav (advisor)
This Bachelor´s thesis is focused on the exploration of polyelectrolyte-surfactant system by fluorescence spectroscopy with pyrene as fluorescent probe. Cationic cetyltrimethylammonium bromide (CTAB) was used as a surfactant. Native hyaluronan was used as polyelectrolyte. Hyaluronan was used in one constant molecular weight and one constant concentration. The whole system was prepared in an aqueous environment. Interactions were studied in area between critical aggregation concentration (CAC) and critical micelle concentration (CMC). The influences of the concentration of pyrene and the effect of the way of the samples preparation on aggregation behavior were also studied. It was found that above the critical aggregation concentration exists an area where stable properties of aggregates can be found. The concentration of pyrene has a major effect on the fluorescence properties of the system. Preparation of samples affects the aggregation of the system. At higher surfactant concentrations, above the stable area, was observed phase separation and gel formation.
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Influence of pH on polyectrolyte-surfactant system
Mourycová, Jana ; Omelka, Ladislav (referee) ; Pekař, Miloslav (advisor)
In this bachelor thesis, was studied influence of pH on aggregation polyelectrolyte-surfactant system using fluorescence spectroscopy and pyrene as fluorescent probe was used. As polyelectrolyte, hyaluronan with different molecular weight was used. Two surfactants cationic surfactant cetyltrimethylamonium bromide (CTAB) and nonionic surfactants polyoxyethylene (20) (TWEEN 20) were selected for interaction with hyaluronan. In the pH range from 3 to 10, no signifact changes in the aggregate non-polar core was observed. This mean system is stable against changes in the observed pH range. At system with higher concentrations of cationic surfactant (CTAB) and higher molecular weight hyaluronan occurred phase separation and gel exclusion was observed.
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Hyaluronan ionocomplexes for cosmetic and farmaceutical applications
Cimalová, Jana ; Obruča, Stanislav (referee) ; Pekař, Miloslav (advisor)
This bachelor thesis is focused on the study of a system of cationic surfactant – hyaluronan. Using fluorescent spectroscopy the critical micellar concentration of Carbethopendecinium bromide (Septonex) was determined in three environments – water, 0,15 M NaCl, and buffer PBS, with three fluorescent probes – pyrene, perylene, and nile red. It was found that the physiological environments (buffer and NaCl) reduce the value of CMC about one order. During the experiments with native hyaluronan, precipitate and gel formation were observed. The following part dealt with aggregate behaviour of the new biologically acceptable amphoteric surfactant tetradecylfosfocholine (TPC) in the environment of water and NaCl according to the increasing molar molecular weight of hyaluronan. As a fluorescent probe perylene was used. These measurements showed that the molar molecular weight or a change of ionic force has no effect on the value of CMC and therefore there are no interactions with hyaluronan. The last surfactant to study was 1,2-dihexadecanoyl-3-dimetylammonium-propan (DPTAP), which is water insoluble. DPTAP is soluble in chloroform, but after evaporation of chloroform and after addition of hyaluronan, it precipitated. The results so far, have confirmed that in terms of research of aggregation by fluorescence spectroscopy the new cationic surfactants TPC and DPTAP are not suitable for complexation with hyaluronan.
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Interaction in ionic surfactant/non-ionic surfactant/polyelectrolyte system
Pihíková, Dominika ; Sedláček, Petr (referee) ; Mravec, Filip (advisor)
The interactions in an ionic surfactant/non-ionic surfactant/polyelectrolyte system have been studied in this bachelor thesis. The value of critical micelle concentration of cationic didecyldimethylammonium chloride (DDAC) has been determined in three different mediums by fluorescence spectroscopy using Pyrene, Perylene and Nile red as probes. The CMC value of non-ionic octylphenol ethoxylate (Triton X-100) has been determined using Pyrene and Perylene as probes. The aggregation behavior of non-ionic and ionic surfactant with addition of polyelectrolyte poly(sodium4-styrensulfonate) (PSS) and native hyaluronan in aqueous solution has been investigated using Pyrene as a probe. Aggregation number in system of ionic/non-ionic surfactant has been determined by a fluorescence quenching with cetylpyridium (CPC) as a quencher and Pyrene as a fluorescence probe. Interactions in ionic surfactant/non-ionic surfactant/polyelectrolyte system were observed. This result was proved by double sigmoid Boltzman´s type curve. The addition of low molecular as well as high molecular hyaluronan had no significant effect on CMC value of the system.
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Study of aggregation in a biopolymer-surfactant system at low surfactant concentration.
Stiborský, Filip ; Vojtová, Lucy (referee) ; Mravec, Filip (advisor)
In this bachelor thesis, the interaction between hyaluronate and surfactant using fluorescent spectroscopy and pyrene as fluorescent probe was studied. A cationic surfactant cetyltrimethylammonium bromide and a native hyaluronate with different molecular size was used. Before a detected critical micelle concentracion, there were no interactions observed in phosphate buffer. However, we could observe electrostatical interactions in the water systém measuring. At low concentration of surfactant (between critical aggregation concentration and critical micelle concentration), the production of gel phase appeared.
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Influence of the temperature on hydrophobic domain quality in a biopolymer-surfactant system.
Hnyluchová, Zuzana ; Kopuletá, Ema (referee) ; Mravec, Filip (advisor)
The temperature dependence on polarity has been investigated by fluorescence spectroscopy with pyrene as a probe. Pyrene was chosen thanks to its unique sensitivity to polarity of the medium. As a biopolymer was chosen 73 kDa and 1,4 MDa hyaluronic acid. Emission spectra of pyrene were measured as a function of temperature in aqous solution of CTAB in a presence and absence of sodium chloride. The results show, that the addition of specific concentration of hyaluronic acid to the sample with CTAB, mostly doesn´t have effect to change increasing/decreasing slope of polarity with the temperature.
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Influence of the preparing of a biopolymer-surfactant system on its aggregation behavior.
Pilgrová, Tereza ; Sedláček, Petr (referee) ; Pekař, Miloslav (advisor)
To study the effect of the method of preparation of biopolymer-surfactant on the aggregation properties several experiments were designed. Closer investigation was made on native hyaluronan-cetyltrimethylammonium bromide (CTAB) system. The system has been studied in the phosphate buffer in aqueous and saline solutions. The effect of the order of substances addition in the samples, the influence of different methods of preparation of certain concentrations of the solutions or the effect of dissolving powdered biopolymer and surfactant on the contrary, the resulting system were investigated. Fluorescence intensity was measured for all prepared samples and its changes were investigated. Pyrene and Nile red were used as fluorescent probes. Some samples were also measured by tensiometer. Measurement of surface tension was based on the method of divorcing the ring from the surface. It was found that the order of the mixing of solutions has a significant effect on the aggregation properties of the system and its appearance. Further effects were detected on the prepared samples by dissolving powdered biopolymer surfactants and vice versa, and environmental influences of the polarity of the system. It was found that sodium chloride abolished the effects of interactions in the system. Effect of method of preparation of certain concentration of CTAB in phosphate buffer is reflected only slightly.
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