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Application of nanoparticles and nanofibers with active compound in cosmetics
Pavelková, Renata ; Obruča, Stanislav (referee) ; Márová, Ivana (advisor)
Presented bachelor thesis is focused on application of nanoparticles and nanofibers with active compound in cosmetics. The theoretical part is aimed at effect of UV radiation on the skin and compounds that are used in photo protection of the skin. The materials and methods used in nanoparticle and nanofiber preparing were characterized. The experimental part deals with preparing of liposomes and marinosomes with active compound. Particles size, stability measured using zeta potential and stability in model environment were observed. Finally, pilot experiments of optimization of nanofibers preparation using electrospinning were carried out.
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Preparation and Characterization of Complex Nanoparticles by Field-Flow Fractionation and Advanced Spectroscopic Methods
Kotouček, Jan ; Krejsek,, Jan (referee) ; Skládal, Petr (referee) ; Turánek,, Jaroslav (advisor)
Liposomes are versatile biocompatible and biodegradable carriers for a variety of medical applications. As the first nanoparticles, they have been approved for pharmaceutical use so far, and many liposome-based preparations are in clinical trials. Classical methods of liposome preparation represent potential limitations in technology transfer from laboratory to industrial scale. New, microfluidic techniques overcome these limitations and offer new possibilities for controlled, continuous preparation of liposomal particles in a laboratory and industrial scale. An important element in the development of new nanoparticle systems is their complex characterization and purification. In addition to the established chromatographic techniques, the Field flow fractionation technique, in particular the Asymmetrical flow Field-flow fractionation, is described. This relatively new technique in conjunction with the MALS/DLS/DAD-UV/dRI online detectors enables the purification and characterization of complex samples. The main advantage of this technique lies in the possibility of separation under native conditions, which plays an important role in the separation of biopolymers in particular. Separation in the “empty” channel then eliminates sample degradation due to unwanted interactions at the stationary phase-sample interface. The theoretical part of this thesis describes the possibilities of preparation, modification, and characterization of liposomal nanoparticles. For this purpose, optical methods based on dynamic light scattering, multi-angle dynamic light scattering and nanoparticle tracking analysis techniques are described, as well as a non-optical method using "particle by the particle" analysis, tunable resistive pulse sensing method. A separate chapter of the theoretical part is dedicated to the technique Asymmetrical flow Field-flow fractionation in connection with the above-mentioned detectors. Important results associated with this work are summarized in the attached scientific paper, together with the result summaries and the author's contributions.
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Encapsulation of vitamins and provitamins into nanoparticles
Martinusíková, Katarína ; Wikarská, Monika (referee) ; Skoumalová, Petra (advisor)
This bachelor thesis is focused on the encapsulation of vitamins in liposomes and the subsequent observation of the properties of this form. The theoretical part is focused on the history of vitamins, their classification into groups, but mainly the general characterization of all vitamins. It describes their function in the human body, the consequences of their deficiencies and also the sources of their income. It also describes the technique of encapsulation in liposomes and their characterization and use in the food industry. The experimental part is focused on the preparation of specific liposomes of vitamins and their mixtures. These prepared liposomes are subsequently characterized by properties such as size, stability and encapsulation efficiency. Long-term stability of prepared liposome particles over 2 and 5 weeks were also investigated. In the conclusion of this thesis are summarized all the results of observations and the subsequent suggestion of the possible use of this technique in food products.
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Use of cell cultures to test cytotoxicity of natural substances
Blažková, Veronika ; Skoumalová, Petra (referee) ; Márová, Ivana (advisor)
This thesis is aimed on study of cell cultures, primarily on introduction of techniques for cultivation of human cells and optimization of their usage in cytotoxicity tests. The thesis is aimed preferably on natural substances and liposomes that are suitable to be used in cosmetics. Theoretical part contains literature review focused on overview of cell cultures, appropriate cultivation conditions for in vitro testing and summary of cytotoxicity tests. Experimental part contains characteristics of natural extracts with antimicrobial effect in terms of content of polyphenols, flavonoids and antioxidant activity. Further, these extracts were encapsulated into the liposomes. Encapsulation effectivity was determined. Distribution of particles size by dynamic light scattering and finally also stability of prepared particles were also determined. Long-term stability in water and gel environment was tested too. This thesis also contains study of optimization of cell culture of human skin keratinocytes cultivation and use of those cells for testing cytotoxicity of selected natural substances by MTT method.
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Influence of liposomal platinum cytostatics on cancer cells – voltammetric study
Laníková, Petra ; Prášek, Jan (referee) ; Hynek, David (advisor)
Aim of this thesis is voltammetric study influence of liposomal platinum cytostatics on cancer cells. One of the goals is summarize available informations about influence of cisplatine on cancer cells, its encapsulation into liposome and affection of this cytostatic cisplatin encapsulated in liposome on cancer cell lines. In literary recherche is detail description of these issues. Than is there specification of voltammetric methods, which serve to electrochemical detection of cisplatin. Based on literary recherche was chosen the best method for detection and subsequently the method was optimalized and than was applied to measuring itself.
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Preparation and stability of core-shell type organic micro- and nanoparticles
Byrtusová, Dana ; Benešová, Pavla (referee) ; Márová, Ivana (advisor)
This bachelor thesis is focused on possibilities of encapsulation polyphenolic compounds in core-shell type organic micro- and nanoparticles. The theoretical part is aimed at characterization of polyphenolic compounds, their biological effects and their possibilities of encapsulation into various types of particles. The experimental part deals with the content of polyphenolic compounds, flavonoids and anthocyanins in guarana, ginseng ang goji. Extracts from these samples were packed into liposomes and polysaccharide particles. Encapsulation´s effectiveness was determined spectrophotometrically. The stability of particles was monitored for size and stability by dynamic light scattering. The particles were exposed to the model body fluids and model food.
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Antimicrobial peptides and their application into foods
Janhuba, Filip ; Petrik, Siniša (referee) ; Márová, Ivana (advisor)
The presented thesis is focused on a study of antimicrobial peptides and their potential applications into foods. Two types of antimicrobial peptides were studied in detail - bacteriocin nisin and antimicrobial enzyme lysozyme. In theoretical part an overview of antimicrobial peptides used in food industry was introduced. The experimental part of this work was focused on optimization of methods for the determination of antimicrobial peptides concentration. Commonly used spectrophotometric methods for quantitative analysis of total proteins as well as high performance liquid chromatography were tested. To determine the molecular weight and purity a modified SDS-PAGE with the addition of urea and tricine buffer was performed. Further, encapsulation of nisin into liposome was tested. Encapsulated particles were prepared by three different procedures: by ultrasonication, ethanol injection and TLE. Particles stability, distribution and size were studied by dynamic light scattering. Stability of encapsulated peptides in model conditions and model foods was studied too. Finally, antibacterial properties of the selected peptides using test system Bacillus subtilis were studied. Nisin already showed antimicrobial effect at low concentrations - about 1 µg/ml. The inhibitory effect of nisin was more evident in liquid cultures.
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Incorporation of low molecular weight and high molecular weight substances into vesicular systems
Geistová, Karolína ; Smilek, Jiří (referee) ; Mravec, Filip (advisor)
This master´s thesis deals with the study of the incorporation of low and high molecular weight substances into liposomal systems. The aim of the work was to determine the encapsulation efficiency (EE) of the active substance and the influence of individual components of the liposomal system on EE. Liposomes were prepared from dipalmitoylphosphatidylcholine. They were stabilized by cholesteroland and phosphatidic acid was added to give a negative charge. Stealth properties gain the binding of polyethylene glycol and other trimethyl chitosan we enabled the entry of liposomes into the bloodstream by the paracellular pathway. Vitamin C and the enzyme bromelain were used for incorporation into liposomes. UV-VIS spectrophotometry was used to determine the encapsulation efficiency of liposomes prepared by combining the individual components. It has been suggested that vitamin C and the enzyme can be incorporated into liposomes, but an enzyme with a higher EE. Furthermore, phosphatidic acid and trimethyl chitosan have been found to affect EE, which increases the EE of vitamin C and decreases the EE of the enzyme.
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Preparation and characterization of nanoliposomal carriers of hydrophobic cytostatics using nanofluidic mixing
Zelníčková, Jaroslava ; Mašek,, Josef (referee) ; Turánek, Jaroslav (advisor)
This diploma thesis is focused on preparation of liposome by relatively new method called nanofluidisation. This method allows the controlled preparation of small unilamellar liposomes in one step. In my thesis I was dealing with optimalization of liposomes preparation which carry hydrophobic cytostatics using this method. Cytotoxic effect of liposomes carrying hydrophobic cytostatics in vitro on cell lines A549 and MCF-7 was determined. In cytotoxicyty test I compared the effect of hydrophobic cytostatics (paclitaxel and derivates of vitamin E specifically alfa-Tos, alfa-TEA) that were incorporated into liposomes prepared via nanofluidisation method and lipid film hydration method. Moreover, a technology of lyophilisation in the presence of cryoprotectants for preparation of liposomes using the method of nanofluidisation was developed.
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Preparation and characterization of food and cosmetic products containing selected vitamins
Kubošová, Jana ; Pavelková, Renata (referee) ; Skoumalová, Petra (advisor)
This bachelor thesis deals with the preparation and characterization of encapsulated forms of lipophilic vitamins in model conditions, focusing on cosmetic and food applications. The theoretical part describes the individual vitamins, their physiological effects, physical and chemical properties, sources and sequels of hypo/hypervitaminoses. Furthermore, their contribution in medicine, food industry and cosmetics, methods of determination and in the last part of the encapsulation method are discussed. Special attention is dedicated to the technique of particles encapsulation using liposomes, which was used in the practical part. The experimental part is focused on the study of liposomes of selected substances of vitamin nature. Namely, the issue of cholecalciferol, tocopherol and -carotene was addressed, for which a suitable method of determination was introduced. Subsequently, encapsulation of these substances and characterization of the prepared liposomes took place. Their size, stability and encapsulation efficiency, which reached high percentages, were determined. Liposomes were further put to the stability tests in digestive juices and model medium simulating cosmetic products. During the measured time, the release of encapsulated substances into the surroundings and their gradual degradation was monitored. At the end of the work, analytes were extracted from real foods and cosmetic products and the measured concentrations were compared with the declared values.
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