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Studying the quality of hydrophobic domains in the phase separated hydrogels
Burešová, Natálie ; Pekař, Miloslav (referee) ; Mravec, Filip (advisor)
The thesis deals with the study of hydrophobic domains in phase separated hydrogels. Two types of polyelectrolytes (hyaluronan, dextran) and oppositely charged surfactants (carbethopendecinium bromide, sodium dodecylsulphate ) are used. Phase separated hydrogels are formed by electrostatic interactions. The target of the thesis is to study the polarity of environment in phase separated hydrogeles by fluorescence spectroscopy. Fluorescence measurements is performed by using pyrene, Nile red and laurdane as fluorescence probes. All probes are sensitive to the polarity of the environment. The results showed that probes can be expected in the polar environment of hydrogels.
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Mixed lipids and their interaction with biopolymers
Cigánková, Hana ; Smilek, Jiří (referee) ; Mravec, Filip (advisor)
This thesis focuses on the study of interactions between hyaluronan and lipid DOTAP and its mixtures with DPPC in water. First, the aggregation behavior of the lipid itself and its mixtures was measured, afterwards the influence of hyaluronan in different concentration was studied. Further the fluorescence anisotropy of fluorescence probe DPH in these mixtures and influence of cholesterol was measured. These properties were investigated by fluorescence spectoscopy using pyrene and DPH as fluorescence probes. The measurements revealed that the addition of hyaluronan has observable influence on the aggregation behavior of the lipid DOTAP and its mixtures with DPPC and that cholesterol influences liposomes fluidity.
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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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Fluorescence quenching in colloid research
Srholcová, Barbora ; Ing. Michal Zavadil, Ph.D (referee) ; Pekař, Miloslav (advisor)
Allready known theoretical observations about utilization of the emission quenching of fluorescence probes in the study of colloids were summarized by finding in the literature search and on the internet, especially the determination of the probe´s location, the aggregation numer and finally the effective viscosity. In the experimental part was studied quenching of the pyrene in the medium of the anionogenic and non-ionic surfactants. As a quenchers were used iodobenzene, tetracyanoethylene and cetylpyridiniumchloride. Selected experiments helped by finding out the aggregation number of the micelle.
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Comparative study of interaction between surfactant and hyaluronan and different polyelectrolytes.
Stiborský, Filip ; Pravda,, Martin (referee) ; Pekař, Miloslav (advisor)
In this diploma thesis, the interactions between polyelectrolyte and surfactant at low and also at high concentration were studied. There was used pyrene as fluorescent probe during the fluorescence spectroscopy measurement, a cationic surfactant cetyltrimethylammonium bromide and as a main polyelectolyte has been chosen sodium polystyrene sulfonate at 1 MDa molecular size. In the medium containing 0.15 M NaCl we could observed a creation of the complexes – precipitates in the surrounding of CMC concentration and behind of this concentration. In the mixtures containing sodium polystyrene sulfonate and hyaluronan together, there was stronger tend to keep aggregation properties of sodium polystyrene sulfonate during difference concentration ratios. Beyond CMC concentration, hyaluronan starts to influence the aggregation properties of the system as well.
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Hydrogels with incorporated proteins
Uhrínová, Alžbeta ; Mravec, Filip (referee) ; Venerová, Tereza (advisor)
This work deals with associative peptides. The aim was to characterize a given associative protein and perform experiments leading to the creation of a hydrogel system containing the selected protein. Casein solutions were initially prepared to determine micellar concentration (CMC) using a pyrene fluorescence probe. Subsequently, the micelles were characterized by dynamic light scattering (DLS) and their size, zeta potential and also the effect of temperature on these properties were determined. Based on the characterization of the particles at temperature, suitable gels were selected into which the protein particles were subsequently incorporated.
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Energy transfer and hydrophobic domains in colloidal systems
Kučerová, Petra ; Čeppan, Michal (referee) ; Pekař, Miloslav (advisor)
In this thesis resonance energy transfer between perylene and fluorescein in tetradecyltrimethylammonium bromide (TTAB) solution was studied. The influence of addition of native or modified sodium hyaluronate on resonance energy transfer was also investigated. The addition of native sodium hyaluronate supports energy transfer at lower fluorescein concentrations and the addition of modified sodium hyaluronate influences the maximal value of energy transfer effectivity. Strong influence on the critical micelle concentration (CMC) values with the addition of sodium hyaluronate during CMC of TTAB determination was investigated. The data indicates not only micelles formation, but also formation of aggregates of sodium hyaluronate with TTAB. Aggregation numbers of TTAB with addition of native and modified sodium hyaluronate by the quenching of pyrene by cetylpyridinium chloride (CPC) was investigated. The addition of sodium hyaluronate into the solution of TTAB changes the average mean aggregation number. Solute exchange between micelles of TTAB and the influence of addition of modified sodium hyaluronate on this exchange was also investigated. No solute exchange between micelles in TTAB and in TTAB with added modified sodium hyaluronate was discovered during this experiment.
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Nanocarriers based on hydrophobized biopolymer
Velcer, Tomáš ; Hnyluchová, Zuzana (referee) ; Venerová, Tereza (advisor)
This work studies the properties of hydrophobically modified biopolymers for potential use in targeted drug delivery. Two samples of hydrophobically modified hyaluronan were selected. The critical micelle concentration (CMC) of these samples was measured using fluorescence spectroscopy, wherein it was found that micelle formation occurs over a wide concentration region. Environment of 0,15 M NaCl causes the decrease of CMC only slightly or not at all. Using the method of dynamic light scattering, particle size was measured. One of the samples exhibited a double size distribution. DLS results are consistent with the results of fluorescence spectroscopy at issue in aggregates forming near the CMC point. Zeta potential measurements provide information about stability of the samples. The results of this study confirm that one of the studied samples seems to be a suitable candidate for the post of drug carrier for targeted delivery.
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Hyaluronane interactions with hydrophobic solutes
Slezáková, Dagmar ; Knotková,, Kateřina (referee) ; Pekař, Miloslav (advisor)
The diploma thesis is based on the study of hydrophobic interactions of the native hyaluronan with selected solutes. On the basis of a literature search were chosen fluorescent probes and fluorescing biologically active substances, which are useful for investigation of colloids as 6-(p-toluidino)-2-naphthalenesulfonic acid (polarity probe), lipophilic vitamin (±)-alpha-tocopherol, pyrene (polarity probe) and finally hydrophilic vitamin riboflavin. In the experimental part of this thesis was studied the influence of solvents with different polarities, or more precisely dielectric constant, on the emission spectra, as well. There were investigated interactions of native hyaluronan with TNS and then interactions, which were influenced by the ionic strength. Such influenced interactions were not observed, that was probably due to the strong solvation´s wrapping of the hyaluronan. Interactions were observed after the process of lyophilisation followed-up by the rehydratation of the samples. For the next study of interactions the riboflavin was chosen and was investigated the REES effect in the native hyaluronan in different concentrations of its different molecular weights. In this case were not observed any shifts in the emission maximum with the excitation wavelenght shift and that is why the interactions of hyaluronan with riboflavin were not demonstrated in the field of chosen concentrations. By using another probe alpha-tocopherol was investigated the associative behaviour of hyaluronan and moreover was observed anisotropy of alpha-tocopherol in different concentrations of different molecular weights of native hyaluronan. The anisotropy reached high values in contrast to the reference solute that was the mixture of glycerol and ethanol. The anisotropy depended more on the molecular weight than on the concentration of hyaluronan. Interactions of hyaluronan were also studied by using the polarity probe pyrene in different concentrations of different molecular weights of the hyaluronan. The pyrene 1:3 ratio did not show the concentration dependence within the chosen concentrations except for the molecular weight 253.9 kg mol–1. Both probes alpha-tocopherol and pyrene were performed by the process of lyophilisation followed-up by the rehydratation, which improved interactions of these probes with hyaluronan.
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Micellar interaction in conditions of phase-separated hydrogel
Lokajová, Aneta ; Sedláček, Petr (referee) ; Mravec, Filip (advisor)
The work aims to study the interaction between hyaluronic acid and the surfactant cetyltrimethylammoniumbromide (CTAB) with proper fluorescent method. Hyaluronan is the strong hydrophilic natur polymer which product viscose solutions with water. It can be found in the joints, skin, eyeball in the live organism. It takes places at cancer growth. It is highly used in cosmetics and medicine because it is nontoxic and biodegradabile. It is used as a aqueous agent or the drug delivery. The work aims to study the interaction with the hydrophobic surfactant with attemption to show or disprove the mutual effect. The emission spectra of pyrene were observed by fluorescence method. The pyren was used as a fluorescent probe.
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