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Use of cell cultures to testing of natural substances of plant origin
Ručková, Michaela ; Kostovová, Iveta (referee) ; Márová, Ivana (advisor)
The presented bachelor thesis is focused on preparing aqueous and oil extracts of plant origin, their characterization and testing of their cytotoxicity and genotoxicity. The theoretical part contains basic information about plants and kinds of spices or tea that were used. Description of used methods of liposome preparation and encapsulation of active substances, measurement of antioxidant activity, particle characterization, cultivation of cell cultures and the principle of cytotoxicity and genotoxicity tests used in this thesis is included. Phenolics and flavonoids content was determined using the spectrophotometry. Antioxidant activity was evaluated spectrophotometrically as well. By ultrasonic homogenization method, liposomes were prepared and active agents were encapsulated into these vesicles. Encapsulation efficiency and stability of active substances were measured by spectrophotometry. By DLS method, size of particles was determined. Particle stability was derived from the zeta potential value. Important part of the bachelor thesis was cultivation of human keratinocytes. The cytotoxicity of extracts and liposomes was assessed by MTT assay on human cell cultures. The genotoxicity of extracts and liposomes was defined using SOS Chromotest on model organism, genetically modified E. coli. In conclusion, the potential use of prepared liposomes in the cosmetic or pharmaceutical industry has been proposed.
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Preparation and use of selected nanoparticles in cosmetics
Hoová, Julie ; Němcová, Andrea (referee) ; Márová, Ivana (advisor)
The Bachelor thesis is focused on preparation and use of selected nanoparticles in cosmetics. The theoretical part deals with general characterization and use of caffeine, phenolics and vitamin C in cosmetics. Further, characterization of liposomes and possibilities of encapsulation of active substances gained from natural sources were introduced. In experimental part liposomes with encapsulated active substances were prepared by three different techniques. Encapsulation efficiency was determined by HPLC/PDA, phenolics spectrophotometrically and vitamin C by titration. Prepared particles were monitored for size and stability by dynamic light scattering. Furthermore, long-term stability of liposomes in model conditions was evaluated as the amount of the active substances released during incubation. In this work was found out that from all tested samples co-encapsulated liposomes are probably the best application form which can be used in face lotion or gel. Co-encapsulated liposomes exhibited relatively high encapsulation efficiency, good stability and thanks to quality of active substances they meet customer’s requirements of cosmetic industry.
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Preparation of encapsulated enzymes for cosmetics application
Bokrová, Jitka ; Čarnecká, Martina (referee) ; Márová, Ivana (advisor)
Presented diploma thesis is focused on testing of an appropriate form of encapsulated enzymes intended for application in cosmetic and pharmaceutical industry. For encapsulation, proteolytic enzymes bromelain, papain and collagenase were used. These enzymes were encapsulated into alginate and chitosan microparticles prepared by an encapsulator and packed into liposomes. Encapsulation effectiveness was evaluated by analysis of total proteins. Particles stability was evaluated in model and real conditions by photometrical analysis of released proteins. Proteolytic activity of released enzymes in model and real conditions were observed too. Alginate and chitosan microparticles prepared by the encapsulator were found as an appropriate form of encapsulated enzymes designed to wound healing. Encapsulation effectiveness of these particles and stability in model conditions were good in comparison with liposomes. Hydrogel and water-oil emulsion were used for analysis of particles stability at real conditions. Hydrogel was found as a good option for preservation of particles as well as proteolytic enzyme activity. Emulsion made particles less stable and proteolytic activity of enzymes decreased rapidly. Encapsulation enables long-term stabilization of biologically active compounds as well as possibility of targeted transport and controlled releasing. Presented diploma thesis suggests possibilities of application encapsulated enzymes in designing more effective formulations for wound healing.
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Use and application of natural antimicrobial substances
Dzurická, Lucia ; Byrtusová, Dana (referee) ; Skoumalová, Petra (advisor)
The presented bachelor thesis is focused on the study of antimicrobial activity of plant substances in combination with bacteriocins and the possibility of creating new encapsulated form of supplement which could be used in food and cosmetic products. 5 plant materials were selected: acerola, cinnamon, cloves, orange peel and yacon syrup. Nisin was chosen as the most suitable bacteriocin. In the theoretical part the components with antimicrobial and antioxidant activity are described, such as the materials of plants and animal origin used in this work. Also the possibility of increasing the antimicrobial effect and possible use of natural antimicrobial substances were described. In experimental part, the aqueous extracts of selected plant materials were firstly spectrophotometrically characterized for the content of total polyphenols, flavonoids and antioxidants. Selected substances were encapsulated into liposomes and subsequently preparation of liposomes coated with chitosan was optimized. The particles were tested for size, short and long term stability, encapsulation efficiency and antioxidant activity. Furthermore, the antimicrobial activities of selected components in nonencapsulated and encapsulated form were observed against four strains of microorganisms (Escherichia coli, Micrococcus luteus, Serratia marcescens and Candida glabrata). Also the expected synergisms of the antimicrobial effects of extracts in combination with nisin and prepared particles were observed.
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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 of organic fibers with the addition of algae extracts
Tuhrinská, Terézia ; Skoumalová, Petra (referee) ; Němcová, Andrea (advisor)
The presented bachelor thesis is focused on the preparation and characterization of organic fibers with an admixture of pre-prepared extracts of microalgae and cyanobacteria containing valuable active compounds. The theoretical part describes selected metabolites of microalgae and cyanobacteria, methods of their analysis, the issue of encapsulation and incorporation of active substances into nanofibers and presents some methods of their preparation. In the experimental part of this thesis, selected strains from the CCALA collection were first cultured. Subsequently, aqueous, ethanol and hexane extracts of biomass from cultured and commercial microalgae and cyanobacteria were prepared. The prepared extracts were spectrophotometrically evaluated in terms of the concentration of chlorophylls, carotenoids, antioxidants, polyphenols, and total proteins present. In addition, 96 % ethanol extracts were subjected to more accurate analysis of chlorophylls and carotenoids by HPLC. The highest antioxidant activity was observed for aqueous extracts. Gelatin nanofibers containing selected extracts were formed by electrospinning method. The formed fibers were examined afterwards to identify a mass fraction of incorporated microalgal and cyanobacterial substances. The fibers were further tested for antioxidant activity with the TEAC assay. The most pronounced antioxidant effect was detected for the fiber with incorporated aqueous extract of the cyanobacterium Arthrospira maxima. Finally, the fibers were tested for their safety in contact with human cells. The amount of fibers used did not show any cytotoxic effect on human keratinocytes and the tested materials can thus be considered safe for application in cosmetics.
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Preparation of food supplements for children containing probiotic bacteria and fruit components
Vetchá, Vendula ; Bendová, Agáta (referee) ; Skoumalová, Petra (advisor)
This bachelor thesis is focused on the preparation of supplementary food with probiotic culture and fruit components. The aim of this work was to determine which of the selected extracts will affect the growth of probiotics and also will contain a certain proportion of phenolic compounds, flavonoids, vitamin C and show antioxidant effects. The theoretical part of the work is focused on the intestinal microbiota, the issue of probiotics and the characterization of encapsulation methods of probiotics. We used extracts of teas and fruit syrups in the experimental part. The antimicrobial activity against model microorganisms Escherichia coli, Micrococcus luteus, Staphylococcus epidermidis and Serratia marcescens was analyzed. Interaction of probiotic bacteria Lactobacillus acidophilus, Lactobacillus casei and Bifidobacterium breve in extracts were tested. After model digestion, cell viability of probiotic bacteria in combination with extracts was examined during and after digestion. In the last part, encapsulated particles containing probiotics and a mixture of tea and syrup were prepared.
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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 encapsulation techniques for production of food for infants
Hoová, Julie ; Čarnecká, Martina (referee) ; Skoumalová, Petra (advisor)
The Diploma thesis deals with use of selected probiotic strains Lactobacillus acidophilus and Bifidobacterium breve in different forms in food for infants. The theoretical part is focused on describing probiotics, encapsulation methods and intestinal gut microbiota of infants. Further, characterization of individual periods of infant feeding and food for infants were introduced. In experimental part the possibilities of encapsulation and lyophilisation of probiotic cells were observed. Probiotic cells were encapsulated into alginate particles. The encapsulator was used for preparation of particles and the most appropriate particles were prepared by encapsulation nozzle with size of 300 µm. Moreover, probiotics viability was monitored by Flow Cytometry, Fluorescence Microscopy and by cultivation (CFU method). Viability of probiotics was monitored during long-term storage in selected food for infants. The appropriate shelf life of non-lyophilized alginate particles in real food have been set at 1 to 2 months. Lyophilized alginate particles could be stored for more than 3 months. Finally, the stability of the particles and viability of encapsulated and non-encapsulated cells in the gastrointestinal tract conditions were also examined. The viabilities of lyophilized cells and cells encapsulated in lyophilized particles were also compared. From the results obtained, non-encapsulated probiotic bacteria cells are more susceptible to negative effects of digestive juices, the percentage of dead probiotic cells after digestion was approximately 80 %. On the other hand, alginate particles showed cell protection from digestive juices, after incomplete cell releasing from particles the percentage of dead probiotic cells did not exceed 20 %. After adequate rehydration, similar results were gained with lyophilized alginate particles. Lyophilized alginate particles have been determined to be the most suitable application form for infants’ food.
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Study of properties and effect of some natural antimicrobial substances
Horňáková, Nikola ; Kostovová, Iveta (referee) ; Márová, Ivana (advisor)
This bachelor thesis is focused on the testing of natural antimicrobial substances of plant origin, dealing with their characterisation, their effect against the microorganisms, optimization of antimicrobial testing and increasing of efficiency of their active compounds. Theoretical part of this thesis describes a characterisation of used natural species, their important properties, spectrophotometrical methods applied in determination of active compounds, opportunities of stabilization of natural substances and various techniques of antimicrobial testing. In experimental part 8 kinds of tea and non-tea plants (Pai Mu Tan, Formosa Gunpowder, Gunpowder Black, Honeybush, Lapacho, Aspalathus linearis and Paullinia cupana) were used and combined with 6 herbal species (Syzygium aromaticum, Urtica dioica, Hippophae rhamnoides, Matricaria recutita, Plantago lanceolata and cinnamon), extracted by different solvents (water, ethanol and dimethyl sulfoxide). In these extracts amount of antioxidants and effect to three bacterial cultures (Escherichia coli, Micrococcus luteus and Serratia marcescens) were determined. To improve antimicrobial, antioxidant activity and stability aqueous tea extracts were encapsulated into liposomal particles.
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