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Native hyaluronan as a delivery system for hydrophobic drugs
Černá, Eva ; Mravec, Filip (referee) ; Pekař, Miloslav (advisor)
The aim of this paper is to discover whether it is possible to use the native form of hyaluronic acid as a hydrophobic drug carrier for a targeted distribution in the body. In its structure, hyaluronic acid is a linear high molecular weight biopolysaccharide which is found in most living organisms. Hyaluronan is involved in many physiological processes and therefore is essential for the functionality of the human body. It is in most tissues of the human body, high concentration is in the skin, the vitreous body and is also observed in cancer cells that contain several receptors for hyaluronan. These receptors include CD44 and RHAMM. The interaction of the hyaluronic acid delivery system and the hydrophobic medicinal with these receptors could ensure a free passage for drugs to the affected tissue, where the release of the drug would destroy the affected cells. The drug would directly target the damaged tissue and did not burden the rest of the body like the cytotoxic agents do. In this paper the native form of hyaluronic acid, which we normally find in the human organism, was chosen as the carrier. Its properties do not stand above other carrier systems, but its biocompatibility and biodegradability in the body greatly exceed them. High molecular weight hyaluronic acid was used as a carrier and the hydrophobic dye sudan red G, a substance of similar properties, was used instead of a hydrophobic drug.
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Interactions between hyaluronan and cetrimide
Adamcová, Zuzana ; Sedláček, Petr (referee) ; Mravec, Filip (advisor)
Interactions between hyaluronan and cationic Cetrimide were studied using fluorescence spectroscopy and turbidimetry in model physiological solution (0,15 M NaCl) and water. Results obtained in water showed strong electrostatic interactions before reaching CMC of the surfactant, which were screened in presence of salt due to high ionic strenght of the solution. Behavior of Cetrimide-hyaluronan systems was compared with the pure TTAB in equivalent system. It was found, that changes in Cetrimide-HA system occure at lower concentrations of the surfactant than in TTAB-HA system. This was probably caused by presence of small amount of CTAB in Cetrimide.
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Influence of conditions on hydrogel preparation
Heger, Richard ; Kalina, Michal (referee) ; Mravec, Filip (advisor)
This work is focused on testing the influence of conditions, more precisely pH, during the creation of hydrogels that were prepared through the hydration of powdery mixtures, by another name solid-mixture way. The hydrogel system was made from a polyelectrolyte, more accurately sodium hyaluronate and a cationic surfactant, CTAB. The influence of pH was studied through observation and rheology. For measuring purposes, the hyaluronan with the molecular weight of 600-800 kDa was used. The system was examined via a series of buffers with the pH of 3,5-11. The gels were dyed by Sudan III for better observability and description. It was found that the best way of adding an active substance was by adding a dye directly into a powdery mixture. The observed properties of the gels didn’t change even after the gels were modified to the ionic strength of a physiological saline solution (0,15 M).
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Time-temperature superposition principle used for study of rheological properties of polymer materials
Kadlec, Martin ; Jarábková, Sabína (referee) ; Smilek, Jiří (advisor)
This bachelor thesis provides a study of the “time-temperature superposition” principle and its applicability to hydrogels. According to current research, honey was chosen due to its simple viscoelastic properties (Newtonian liquid) as a suitable material for optimalization procedure. The knowledge obtained during optimization was further applied to real hydrogel materials, namely the agarose gel was selected as an example of thermoreversible hydrogel, as well as the hyaluronan and dextran gels as examples of hydrogels formed by the interaction of the polyelectrolyte with the opposite charged surfactant. By using the “time-temperature superposition” principle, a considerable increase in the range of the observed oscillation frequencies was achieved for all chosen samples, which led to gain of information about storage and loss or complex modulus, which could not be achieved by classical oscillatory test. Although master curves were generated for all the samples examined, curves for storage and loss modulus could be created separately only for hydrogels formed by the interaction of the polyelectrolyte with the opposite charged surfactant. For this reason, this group of hydrogels appears to be more suitable materials for the application of the “time-temperature superposition” principle.
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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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Analytical methods for hyaluronan
Černá, Lucie ; Mravec, Filip (referee) ; Pekař, Miloslav (advisor)
This thesis deals with of simple methods for determining the concentration of hyaluronan in aqueous solution and compares the performance of each selected methods such as UV -VIS spectroscopy, IR spectroscopy and the method of determination by alcian blue. These methods were chosen for their simplicity and low cost of the assay. As the most reliable of these three methods, seems to be the method of IR spectroscopy. Determination of the concentration of HA by this method is relatively accurate at concentrations above 0.4 % in deionized water. The method of UV-VIS spectroscopy is based on a linear calibration curve determination, but no characteristic peak was detected, which means that this determination could be influenced by other components of the aqueous solution. Weak ionic strength and molecular weight of HA does not affect the determination. In the method with alcian blue, the results were quite unsatisfactory and does not correspond with the literature. Therefore, this method is not applicable in this form for determining the concentration of HA. Another option for determining the concentration of the HA is ELISA method. These are commercially produced kits suitable for determining low concentrations in low volumes. Its price is higher than the above- mentioned methods.
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Optimizing the determination of distribution of surfactant at phase separation in polymer-surfactant system
Sátorová, Kateřina ; Pekař, Miloslav (referee) ; Mravec, Filip (advisor)
This diploma thesis is focused on the determination of distribution a surfactant at phase separation in polymer-surfactant system. Sodium hyaluronate of three molecular weights was chosen as a polymer, CTAB was used as a surfactant. The experiments were performed in an environment of 0,15 M NaCl. The measurement of the CTAB concentration in samples was based on the formation of coloured complexes of CTAB and picric acid in chloroform. UV-VIS spectroscopy was used for the detection of these complexes. The content of CTAB was determined in four gels of different composition (2% Hya + 200 mM CTAB, 2 % Hya + 50 mM CTAB, 0,5 % Hya + 200 mM CTAB, 0,5 % Hya + 50 mM CTAB). All gels were gradually washed five times with 0,15 M NaCl. The content of CTAB was determined for newly prepared gels and for gels after 2 and 4 months since the preparation. Initial concentrations of CTAB and hyaluronan have greater influence on the distribution of CTAB after phase separation, than the molecular weight of hyaluronan. The content of CTAB in newly prepared gels and after 4 months since preparation is very similar and demonstrates the stability of the system.
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Interactions between Hyaluronan and Surface Active Substances
Krouská, Jitka ; Burgert,, Ladislav (referee) ; Lehocký,, Marián (referee) ; Vikegard,, Peter (referee) ; Pekař, Miloslav (advisor)
Vliv hyaluronanu na micelizaci tenzidů byl studován různými fyzikálně-chemickými metodami. Byly zvoleny dva kationaktivní tenzidy, a to tetradecyltrimethylammonium bromid (TTAB) a cetyltrimethylammonium bromid (CTAB). Metoda izotermické titrační kalorimetrie byla využita pro stanovení entalpie micelizace, tenziometrie popisuje povrchové vlastnosti daných vzorků. Byl sledován také vliv různé molekulové hmotnosti použitého hyaluronanu a délka alkylového řetězce tenzidu na agregační chování daného systému. Výsledkem jsou hodnoty kritické micelární nebo agregační koncentrace tenzidu. V neposlední řadě se diskutuje využití agregátů hyaluronan-tenzid jako možné nosiče pro cílenou distribuci léčiv.
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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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