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Study of interaction of charged catanionic vesicles with oppositely charged polyelectrolytes
Havlíková, Martina ; Lehocký, Marián (referee) ; Táborský, Petr (referee) ; Mravec, Filip (advisor)
The presented dissertation is focused on positively charged catanionic vesicles and their interactions with oppositely charged hyaluronan. The catanionic vesicles were composed of ion pair amphiphile of hexadecyltrimethylammonium-dodecyl sulfate (HTMA-DS), a positively charged double-chain surfactant that electrostatically stabilized the vesicles and cholesterol. Cholesterol may strongly affect the physico-chemical properties of the vesicles, therefore it was necessary to determine its optimal amount in the membrane. Especially, the size and stability of vesicles were studied by dynamic and electrophoretic light scattering, membrane microviscosity by fluorescence anisotropy and hydration of the outer part of the membrane by generalized polarization. It was found that vesicles show ideal properties at a cholesterol content about 40 mol.%. These catanionic vesicles interacted with hyaluronan regardless of its molecular weight. Visible aggregates were observed around their isoelectric point, but they disintegrated with increasing polymer concentration. However, this decay was different for different hyaluronan molecular weights. An increase in ionic strength, or more precisely concentration of sodium chloride above 25 mM had a destabilizing effect on both uncoated and hyaluronan coated vesicles. It was also interesting that increasing the ionic strength affected the microviscosity of the membrane, which became more fluid, but there were no significant changes in the hydration of its outer part. The presented work is also focused on the preparation and characterization of a new type of catanionic vesicles, where the cationic surfactants HTMAB was replaced by Septonex. The same methods were chosen for their characterization as in the case of the HTMA-DS vesicles. This exchange of surfactants had a major effect on the resulting properties of the vesicles, which are stable even at very low cholesterol content. In accordance with long-term stability, a suitable concentration of cholesterol was chosen from 15 mol.% and higher.
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Hydrogels with incorporated enzymes
Geistová, Karolína ; Krouská, Jitka (referee) ; Mravec, Filip (advisor)
This bachelor thesis deals with the study of incorporation of enzymes into phase separated hydrogels. The aim of this work is to determine the enzyme activity in phase separated gels. Gels were prepared by the dry-way based on the interaction of negatively charged polyelectrolyte (hyaluronan) with positively charged surfactant (Septonex). Two enzymes, bromelain and collagenase, were incorporated into the hydrogels. To determine enzyme activity, the modified albumin protein with bound sulfanilamide group (azoalbumin) was used as a substrate. The enzyme activity of the enzyme itself, the enzyme activity affected by one of the two components of the system as well as the activity of the enzyme directly in the hydrogel was determined on UV-VIS spectrophotometry. The enzyme was found to be incorporated in the hydrogel. Furthermore, a significant effect of the positively charged surfactant on the enzyme activity was detected in phase-separated hydrogels.
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Advanced Fluorescence Techniques in Research on Micellar Systems and Their Interactions with Biopolymers
Holínková, Petra ; Burgert, Ladislav (referee) ; Táborský, Petr (referee) ; Pekař, Miloslav (advisor)
The dissertation thesis deals with study of advanced steady-state and time-resolved fluorescence techniques, which can be used for study of micellar systems properties. Selected fluorescence techniques were used for characterization of Septonex and CTAB cationic micellar systems and theirs interactions with hyaluronan. Fluorescent probe pyrene was used for determination of critical micelle concentration (CMC) and micellar aggregation number of these surfactants. The changes of fluorescence behaviour of fluorescein and prodan were studied in wide concentration range of Septonex. Next chapter of thesis deals with study of Förster resonance energy transfer between perylene and fluorescein in Septonex and CTAB micellar solutions and the effect of hyaluronan addition to these systems. Also steady-state and time-resolved fluorescence anisotropy studies were used for research of the effect of hyaluronan addition to micellar solutions. The last chapter of this thesis is focused on photophysical behaviour of Prodan in different solutions (water, Septonex solutions below CMC, hyaluronan solution, Septonex micellar solution and Septonex micellar solution with hyaluronan), which was discussed on the basis of time-resolved emission spectra.
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Preparation and characterization of magnetic nanoparticles modified by surfactants
Solnička, Ondřej ; Másilko, Jiří (referee) ; Enev, Vojtěch (advisor)
Theme of this bachelors thesis was preparation and characterization of iron(II, III) oxide nanoparticles. Those nanoparticles were modified by surfactants. The surfactants which were used for modifiation were CTAB, TTAB and Septonex. Prepared and modified nanoparticles were taken for characterization by using methods of elemental analysis, thermogravimetry, dynamic light scattering and transmission electron microscopy (TEM). Results from each measurement shows that nanoparticles modified by CTAB had on average 85.9 nm in diameter, TTAB size was on average 108.3 nm and the last nanoparticles with Septonex had 246.6 nm in diameter on average. Prepared modified nanoparticles except the Septonex ones were stable enough to not aggregate. Septonex modified nanoparticles had greater tendenty to aggregate.
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New amphiphilic ion pairs for the preparation of vesicular systems
Strnadová, Martina ; Kalina, Michal (referee) ; Mravec, Filip (advisor)
The bachelor thesis is focused on the preparation of the new vesicular systems. The aim of this work is to create the most stable system and find out the most optimal working method for its production. The compared parameters were the energy used for dispersion, the molar percentage of cholesterol added, and the type of double-stranded positively charged surfactant. The whole system consisted of negatively charged surfactant sodium dodecyl sulfate and positively charged surfactant septonex. Each system was compared over time to determine stability based on the size of the particle, zeta potential, and turbidity of the solution. By the method of dynamic light scattering, particle size and polydispersity coefficient were measured. On the other hand, the zeta potential was determined by electrophoretic light scattering. The result of the thesis presents the most suitable parameters of ultrasonic dispersion, the molar proportion of cholesterol, and the use of cationic surfactant. Based on these parameters, it is possible to create a physically stable system for a long period of time, thanks to which such a system can be used in the future for other applications.
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Diffusion of dyes in biopolymeric hydrogels
Vyroubal, David ; Mravec, Filip (referee) ; Klučáková, Martina (advisor)
This thesis is focused on diffusion of dyes in biopolymer-based hydrogels. These hydrogels are based on interaction between biopolymer-like electrolytes with oppositely charged surfactants. When polyelectrolytes interact with oppositely charged surfactants, micelle-like nano-containers can be formed. These nano-containers are able of binding hydrophobic compounds. In this study, combination of modified dextran (diethylaminoethyl dextran) with positive charge and oppositely charged sodium dodecylsulphate as surfactant was used for preparation of hydrogels. Next type of hydrogel was based on hyaluronan and positive charged surfactant Septonex (carbethoxypendecinium bromide). As a diffusion probes in hydrogels dyes Nile red and ATTO 488 were used. The diffusion of these dyes from aqeous solutions of NaCl or surfactants into hydrogels was monitored in time. Transport of dyes into structure of hydrogels was characterized by diffusion coefficients and structural parameters of hydrogels.
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Characterization of coloid particles by excited-state proton transfer with advanced fluorescence techniques
Kotouček, Jan ; Mravec, Filip (referee) ; Pekař, Miloslav (advisor)
The deprotonation characteristics of fluorescent probes -naphthol and 8-hydroxypyrene-1,3,6-trisulphonic acid (HPTS) were studied in this diploma thesis, using steady-state and time-resolved fluorescence spectroscopy. Two cationic surfactants, Septonex and cetyltrimethylammonium bromide (CTAB), were studied. These surfactants were measured in the complex with hyaluronan (1.75 MDa, 1 MDa and 300 kDa). Steady-state fluorescence was used for determination of critical aggregation concentration of each surfactant and pKa*. Time-resolved fluorescence decays were used to calculate the average lifetimes and the deprotonation constants of naphthol and HPTS. The measurement with hyaluronan were compared with the polystyrenesulfonate (PSS) – surfactant system. The effect of hydration shell of hyaluronan on hyaluronan – surfactant complex formation results from the comparison of above mentioned systems. Large differences were found in the deprotonation characteristic between surfactants and even between individual molecular weights of hyaluronan. The measurement shows that the hydration shell is located near to the dissociated carboxyl groups of hyaluronan chain, where the interaction with the positively charged surfactants occurs. Furthermore, the aggregation number of Septonex was determined by quenching of pyrene using cetylpyridinium chloride (CPC) as a quencher. The aggregation number for 20 mM Septonex solution was determined as a value of 104 molecules. CPC was used for confirmation of the localization of -naphthol in the micelles of CTAB and polymer – CTAB, respectively.
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Hyaluronan ion complexes
Cimalová, Jana ; Sedlařík, Vladimír (referee) ; Pekař, Miloslav (advisor)
This diploma thesis is focused on the study of physical and chemical properties of hyaluronan and cationic surfactant. As the cationic surfactant Septonex was used. The influence of the environment on the system, the effect of molecular weight of hyaluronan, and its concentration was studied. Then, the study of the influence and the effects of concentration of Septonex on the interaction of hyaluronan-surfactant followed. Different methods of measurement were chosen to characterize these ionokomplexes. The critical micelle concentration of the surfactant itself was measured, and then also with the addition of hyaluronan by spectrofluorimetry with fluorescent probe pyren. It was found, that hyaluronan forms gel with Septonex. On this basis, gels were prepared for three different molecular weights of hyaluronan – 300 kDa, 806 kDa and 1697 kDa. Gels were prepared in a ratio of hyaluronan – surfactant 1:1. In gels prepared in this way, the influence of environmental water and 0.15 M NaCl was studied and it was found that at 0.15 M NaCl clear gels are formed. Selected samples of the gels were then measured with oscillatory testing and the rheological behavior of gels of Septonex was studied. As the last method the turbidimetric measurement was chosen, which characterized the turbidity point in the gradual addition of Septonex to sodium hyaluronate solution. Again, the effect of the molecular weight of hyaluronan and its concentration in two environments - water and 0.15 M NaCl was evaluated. It was found that 0,15 M NaCl suppresses formation of turbidity and formation of precipitates.
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Structural aspects of interaction between humic substances and charged organic xenobiotics
Prisažný, Adam ; Smilek, Jiří (referee) ; Sedláček, Petr (advisor)
This diploma thesis was focused on studying the interaction of humic substances with ionic organic xenobiotics and its structural aspects. The method was chosen from my bachelor thesis, steady-state fluorescence spectroscopy, which is suitable for substances with weak fluorescence. The results showed that the interaction between humic acids and representatives of ionic organic xenobiotics (Septonex) was reflected in fluorescence quenching of humic acids and the shift of emission maximum to lower wavelength, hypsochromic (blue) shift. From the measurement results, we can assume that the interaction that is formed between the aromatic structures in humic acids and Septonex could be -cation interaction.
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Influence of molecular weight of polymers on phase-separated hydrogels
Kolomá, Nikola ; Smilek, Jiří (referee) ; Mravec, Filip (advisor)
This work is focused on study of individual systems of hyaluronan mixtures. All mixtures of hyaluronans they were ready at molecular weights of 15-30 kDa, 80-150 kDa, or 1500-1750 kDa. These individual molecular weights were combined in different ratios most often in ratios of 70:30, 50:50 and 30:70. Septonex surfactant was used for all mixed samples and CTAB was used for last experiment too. At first the gel samples were prepared by a dry route of individual powders with accurately calculated batches. All solutions were prepared wet route with stock of exactly calculated concentration. Physical gels were prepared by combinations of Septonex and hyaluronan with the different molecular weights. In solution formed irregular shapes which we could describe as "marbles" or "beads". After the sample was to stand, the individual „marbles“ of the gel were formed whole mass. Was done too many experiments with physical gels, which has been carried out behavior of mixtures hyaluronans with different molecular weights. All physical and chemical properties were observed by the eye, described and in the and were photographed. Was used hydrophil dye in experiments and served for better visualize comparison samples between themselves. The last experiment was rheological measurement with which is possible characterize viscosity od individual samples.
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