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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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Microrheology with Fluorescence Correlation Spectroscopy
Kábrtová, Petra ; Sedláček, Petr (referee) ; Mondek, Jakub (advisor)
A comparison of three passive microrheological techniques was made with the emphasis on FCS. Fluorescently labelled and unlabelled polystyrene particles were used to probe a microrheological response of glycerol solutions, Mili-Q water and sodium hyaluronate solutions. In addition, for FCS technique an approximated equation for calculation of MSD values was derived and verified. It was found that FCS outmatches current microrheological techniques of DLS and video-based particle tracking by its ability to gain a broader data range including the area in which, until now, it was impossible to describe a microrheological behaviour of samples reliably.
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Study on relaxation properties of hydrogels using rheological techniques
Lorenc, Pavel ; Klučáková, Martina (referee) ; Smilek, Jiří (advisor)
Submitted bachelor’s thesis deals with the study of relaxation and time stress of viscoelastic substances (hydrogels) using selected rheological techniques, especially with regard to oscillation tests. Hydrogels with different types of crosslinking were prepared and their properties were compared during the measurement. Agarose hydrogels (physically), alginate hydrogels (physicochemically), polyvinyl alcohol hydrogels (chemically) and hyaluronic acid hydrogels (ionically). In this bachelor's thesis, methods for measuring the relaxation properties of these hydrogels using a rheometer were designed and subsequently applied. We examined the relaxation properties of hydrogels using time-dependent tests that examined changes in amplitude over time. We performed a test that had 3 intervals, in which we monitored the response of hydrogels to the changes by changing the amplitudes of deformation with time. The relaxation properties of these hydrogels were studied, which showed significant differences in the behavior of hydrogels in measurements.By these methods, it was found that the structure of hydrogels and concentrations have an influence on relaxation. From the measured results, it was shown that the best relaxing properties have chemically crosslinked polyvinyl alcohol hydrogel, the lowest ability to relax was shown by physico-chemically crosslinked alginate hydrogel.The results from the measured data were finally compared, assessed and commented on the differences between the individual hydrogels.
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Viscosimetry of hyaluronan-amphiphile systems
Trojan, Martin ; Běťák, Jiří (referee) ; Chytil, Martin (advisor)
This bachelor thesis investigates the interaction between the polysaccharide sodium hyaluronate (HA) and some amphiphilic molecules. As amphiphilic molecules the aminoacids lysine and 6 – aminocaproic acids were chosen. The presence of the carboxylic group on HA and the aminogroup on the aminoacids suggests electrostatic interaction between these two compounds. This supposal offers the possibility to modificate HA and uses it as a new type of a carrier of bioactive compounds, for example medicals. The HA of two molecular weights (1.75MDa and 70kDa) were chosen for the study of the interactions with the aminoacids. Solutions of HA and stock solutions of the aminoacids were prepared for their mutual mixing in the various volume ratios. The solutions and the mixtures were measured in three types of environment: pure water, 0.15M solution of NaCl and in phosphate buffer (PBS) of pH 6. The interactions were investigated by means of reometry. Our results show, the presence of interactions between HA and the aminoacids mainly in the system consisting the 1.75 MDa HA and lysine in water. As a result of the interactions, a decrease in the viscosity of the mixtures comparing to the HA solutions was observed.
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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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Hydrogels with incorporated vesicular systems
Kalendová, Lucie ; Pekař, Miloslav (referee) ; Venerová, Tereza (advisor)
This bachelor thesis deals with the incorporation of vesicular systems into polysaccharide – based hydrogels. The chosen vesicular systems were Ion Pair Amphiphile vesicles (IPA), which consist of cetrimethylammonium bromide, sodium dodecyl sulfate, dioctadecyldimethylammonium chloride and cholesterol. The work is divided into two parts. In the first part, interactions between IPA vesicles and sodium hyaluronan and interactions between IPA vesicles and sodium polystyrene sulfonate were studied. Even though interactions occured, it was not possible to prepare a hydrogel based on these interactions. The second part of this work deals with incorporation of IPA vesicles into an agarose – based gel when heated. Particles of two different concentrations (1 mmol·l1 and 2 mmol·l1) were used. The effect of the IPA particles on the properties of the gel was observed with rheological measurements. The results showed that the gel with incorporated particles has a longer linear viscoelastic region and that there were no interactions between the IPA particles and the gel. The particles only fill the gaps of the gel network. Different behaviour within the gels with different concentrations of IPA vesicles was not observed.
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Hyaluronan effect on transdermal penetration of selected pharmaceutical substances
Ureš, Tomáš ; Sedláček, Petr (referee) ; Pekař, Miloslav (advisor)
This work deals with the transmittance of a family of non-steroidal anti-inflammatory drugs across biological membranes in combination with the use of hyaluronan. Hyaluronic acid (hyaluronan, HYA) is a linear polysaccharide formed from disacharide units containing N-acetyl-D-glucosamine and glucuronic acid. HYA is present in almost all biological fluids and tissues, so there is an assumption that could affect the penetration of substances through the skin. Standards were prepared by anti-inflammatory drugs in admixture with various concentrations hyaluronan and subsequently measured transmittance of such substances through the skin. Standards were applied to the skin obtained from pig auricle. The drug content was determined by HPLC.
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Wound dressing nanofibers mats fabricated from nanocomposite material
Čileková, Marta ; Pavliňák,, David (referee) ; Abdellatif, Abdelmohsan (advisor)
Boli pripravené kryty rán na bázi prírodných látok polyvinyl alcohol/ hyaluronan/ strieborné nanočastice (PVA/ HA/ Ag-NPs). Hyaluronan bol použitý ako redukčné a stabilizačné činidlo pre syntézu nanočastíc striebra. Pri príprave Ag-NPs boli testované viaceré parametre ako koncentrácia dusičnanu strieborného ako zdroja Ag-NPs (0,01; 0,1;0,5;1 M), koncentrácia kyseliny hyalurónovej (1,2 %) a jej rozdielna molekulová hmotnosť. Kryty rán z nanovlákien boli pripravené pomocou techniky electro-spinning z roztokov líšiacich sa pomerom PVA a HA/Ag-NPs (100; 90/10; 80/20; 60/40; 50/50). Vlastnosti nanokompozitu HA/Ag-NPs boli hodnotené pomocou TEM, reológie, DLS, XRD, UV/Vis spektroskopie a kryty rán boli charakterizované pomocou SEM, TGA, FTIR a ťahovej skúšky.
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SEC-MALLS study on hyaluronan-albumin interactions
Sereda, Alena ; Kalina, Michal (referee) ; Chytil, Martin (advisor)
This work is focused on interactions of hyaluronan (HA) and bovine serum albumin (BSA). These interactions were studied by SEC-MALLS method, where molar mass, root mean square (rms) radii, hydronamic radius and intrinsic viscosity of particles of the system were defined. Additionally, interactions were examined by measurement of DLS and zeta potential, where hydrodynamic radius and a value of the zeta potential were defined. The interactions were confirmed at any of the used media, but with different efficiency. Furthermore it was found out, that the high ionic strength minimizes the range of the interactions, magnifies the radii of the complexes, their intrinsic viscosity and zeta potential. It was proved that complexes become smaller in their radii and viscosity with growing BSA concentraton.
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Interactions and aggregation in hyaluronan-aminoacid-surfactant systems
Venerová, Tereza ; Velebný, Vladimír (referee) ; Pekař, Miloslav (advisor)
The effect of native hyaluronan addition on CMC of excluded surfactants (Tween 20, BETADET THC 2, SDS, CTAT a CTAB) in physiological solution (0,15 M NaCl) has been investigated by fluoresence spectroscopy with Pyrene and Nile red as probes. The greatest influence on CMC has been observed in systems with cationic surfactant. System of cetyltrimethylamonnium bromid has been closely studied. Solubilization experiments with a hydrophobic dye Sudan red has been realized and aggregation number of this system has been determinated via fluorescence quenching with cetylpyridinium chloride as quencher and Pyrene as fluorescence probe. Addition of hyaluronan (native or hydrophobicaly modified) reduces aggregation number of system.
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