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Study of stability of antimicrobial nanomaterials in different environment by HPLC
Janderová, Šárka ; Trachtová, Štěpánka (referee) ; Márová, Ivana (advisor)
This thesis deals with the preparation of nanomaterials (nanoparticles, nanofibers) with encapsulated active substances and the stability of these nanomaterials in different environments using liquid chromatography. The development and optimization of methods for monitoring the kinetics of release of encapsulated substances from nanomaterials into various environments play a key role in research aimed at their utilization in the pharmaceutical, food, and cosmetic industries, and also contribute to the development of new antimicrobial nanomaterials limiting antibiotic resistance. The theoretical part focuses on the characterization of nanomaterials (nanoparticles, liposomes, nanofibers, and combined nanomaterials). Another section of the literature review deals with natural (eugenol, carvacrol, curcumin, thymol, vitamin E) and synthetic (ampicillin, streptomycin) antimicrobial substances. The final chapter discusses the instrumental methods used for the characterization of nanomaterials and the evaluation of their stability, released, and degradation products. In the practical part, the antioxidant activity of selected substances was first monitored. Liposomes were prepared, and their stability and encapsulation efficiency were examined. An HPLC method for determining active substances was optimized. Subsequently, the gradual release of the mentioned active substances from prepared liposomes was monitored. Nanofibers were prepared using forcespinning, and their release into selected model environments was also analyzed. Distilled water with minimal ion and impurity content served as a model environment, as well as a physiological solution at 25°C, simulating the temperature of the human body surface, and a physiological solution at 37°C, characteristic of human body temperature. This analysis provides a comprehensive view of the behavior of prepared materials and their potential practical applications. In the final section, combined nanomaterials (based on nanofibers and nanoparticles) with encapsulated active substances were prepared. The gradual release of active substances and any potential synergistic effects were examined for these nanomaterials.
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Characterization of cytotoxic effect of combined antimicrobial nanomaterials
Kozlíčková, Hana ; Fialová, Lenka (referee) ; Márová, Ivana (advisor)
This thesis deals with the study of the effects of combined nanomaterials on human skin cells. Pure antimicrobial substances, two types of liposomes enriched with antimicrobial substances, nanofibers with antimicrobial substances and, finally, four types of combined nanomaterials were analyzed from the point of view of cytotoxicity. The analysed active substances were eugenol, thymol, cavarcrol, curcumin, vitamin E and the antibiotics streptomycin and ampicillin. In the theoretical part of the work, the cell line of human keratinocytes, used in the experimental part of the work for cell tests, was characterized. Furthermore, individual active substances with an antimicrobial effect were described and the principles of biological effects were described, which include antimicrobial, antioxidant, cytotoxic and synergistic effects. Additionally, the theoretical part described individual nanomaterials, their preparation and usage in cosmetics and medicine. The experimental part was based on the characterization of prepared nanomaterials and on testing the influence of individual antimicrobial substances on the proliferation and viability of human HaCaT cells. Using the DLS method, the size of the prepared liposomes was measured and the effect of PHB and the type of active substance on their size was studied. MTT and LDH tests were chosen to test the cytotoxicity of individual substances. Furthermore, a scratch test was performed to monitor the effect of the investigated substances on proliferation and the rate of wound healing by cells. The last performed tests were immune response assays, in which were tested the samples for production of the human anti-inflammatory cytokines IL-6 and IL-8. In experimental part of this thesis, it was found that the prepared nanofibrous materials are safe for use in healthcare or cosmetics and, in the future, suitable to produce nanofibrous wound coverings enriched with antimicrobial substances, which can give them exceptional properties.
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Preparation and characterization of nanofiber wound dressings
Jiroušková, Pavla ; Uhlířová, Renata (referee) ; Skoumalová, Petra (advisor)
The presented bachelor thesis deals with the preparation and characterization of nanofiber wound dressings. The materials used for the preparation of nanofibers were poly(3-hydroxybutyrate) and gelatin. Active substances, such as ampicillin and ibuprofen, were incorporated into the wound dressings. In the theoretical part of the work skin, wound healing, and nanofibers and their properties were described. This part also deals with the use of nanofibers as wound dressings. Additionally, it depicts materials suitable for the preparation of nanofiber wound dressings. Furthermore, this section contains a review focused on various methods of preparation of nanofibers, drugs that can be incorporated into the fibers and the subsequent determination of the safety of the prepared dressings. In the experimental part, nanofiber wound dressings containing ampicillin and ibuprofen were produced. The maximum possible amount of active substance feasible to be incorporated into the nanofibers has been determined. Subsequently, combined wound dressings were prepared to study the release rate of active substances into the model environments. The release in the time intervals was determined spectrophotometrically. Finally, antimicrobial activity assays were performed to test the effect of dressings containing ampicillin on selected strains of microbiirganism: S. Epidermidis, E. Coli and M. Luteus
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Carbon nanoparticles from South Moravian lignite
Drozdová, Miroslava ; Enev, Vojtěch (referee) ; Pekař, Miloslav (advisor)
This bachelor thesis deals with the issue of isolation of carbon nanoparticles from lignite, which seems to be their cheap source. The aim of this work is to elaborate a research on the use of coal matrices as a source of carbon nanoparticles and also to design and perform further experiments testing the usability of South Moravian lignite. Based on the literature search, a simple mechanochemical procedure was proposed to obtain carbon nanoparticles from lignite. The lignite was mechanically stressed by the ultrasonic disintegrator with simultaneous action of water or hydrogen peroxide. Lignite has been able to isolate particles fluorescing in the blue visible light, but the process will need to be further modified. However, South Moravian lignite could be a source of carbon nanoparticles.
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Preparation and application of combined functionalized nanomaterials with antimicrobial effect in food and cosmetics
Vojteková, Vanesa ; Skoumalová, Petra (referee) ; Márová, Ivana (advisor)
The presented diploma thesis is focused on the preparation of combined nanomaterials that were functionalized with an antimicrobial ingredient. Liposomes were prepared without the addition of polyhydroxybutyrate but also with its addition. The nanofibers were obtained from gelatin solution by electrospinning. Thymol, eugenol, curcumin and vitamin E were used for functionalization. In the theoretical part, individual bioactive components, materials for the preparation of nanoparticles and nanofibers as well as methods of synthesis of these materials were characterized. The applications of the prepared nanoparticles as preservatives in the food industry and applications of nanofibers in wound healing and in the cosmetics industry have also been described. In the experimental part, the antioxidant activity of the active substances was determined. Two types of liposomes were prepared and evaluated for size, stability and encapsulation efficiency. Subsequently, the release rate of active substances from the prepared liposomes was monitored. Nanofibers with combinations of substances that were encapsulated in liposomes were prepared by electrospinning. The antimicrobial activity of the active ingredients, nanofibers and nanomaterials was monitored on three strains of microorganisms, namely Micrococcus luteus, Staphylococcus epidermidis and Escherichia coli. The individual substances in pure as well as encapsulated form were combined with each other to observe the possible synergistic effect. The prepared nanoparticles and nanofibers with antimicrobial components were tested for cytotoxicity on human keratinocytes. The nanofibers were also subjected to a scratch test, which simulates the wound healing process.
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Development and preparation of antimcrobial nanostructure biomaterials
Drabíková, Nela ; Skoumalová, Petra (referee) ; Márová, Ivana (advisor)
The presented diploma thesis deals with the optimalisation of preparation and the preparation of combined nanostructured antimicrobial biomaterials itself. In the theoretical part, a review focused on used materials and consequently preparation of nanoparticles and nanofibers was elaborated. Furthermore, the used antimicrobial substances – curcumin and ampicillin, and the principle of cytotoxicity assay were described.In practical part the optimalisation process is described. Furthermore the safety of prepared materials and used antimicrobial substances on HaCaT cell line was tested, in order to confirm their possible further use in cosmetic and pharmaceutical industry. Great part of the thesis deals with evaluation of the antimicrobial activity of used substances and prepared combined nanomaterials on multiple microorganisms from grampositive bacteria, gramnegative bacteria and yeasts. Also the release speed of antimicrobial substances from prepared nanomaterials was determined by spectrophotometer. The amount of released ampicillin from prepared nanomaterials was determined by liquid chromatography.
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Automatisation of an experimental setup for optical spectroscopy
Hrabina, Jakub ; Gallina, Pavel (referee) ; Dvořák, Petr (advisor)
This bachelor thesis deals with the development and description of an application that enabled full automation of an experimental assembly for spectral mapping using confocal optical microscopy. It presents communication with all individual devices and explains the principle of data collection from the examined sample and their subsequent mathematical processing. The output of this application is the visualization of 2D spectral maps evaluating the intensity and position of spectral peaks, with the possibility of exporting user-friendly point spectra. In addition, the work further describes a research study of optical confocal spectroscopy with applications to nanophotonics. In addition, the functionality of the whole application is demonstrated on the spectral response of a 2D metasurface, which acts as a Bayer mask.
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Nanotechnology utilization in nuclear industry and research
Skalička, Jiří ; Štefánik, Milan (referee) ; Katovský, Karel (advisor)
This thesis introduces reader to current knowledge of nanomaterials and their usage. It summarises production methods and usage of different materials in nuclear power plants, nuclear research and nuclear medicine. Theoretical part of this thesis is dedicated to possible usage of carbon nanotubes for neutron beam collimation and guides. In experimental part different materials were tested in measuring box connected to horizontal radial channel of VR-1 nuclear reactor and their influence on neutron flux was measured. Tested samples were non-oriented carbon nanotubes, carbon nanofibers, alumina nanowires, oriented carbon nanotubes with several angles of rotation and these samples were compared with results of graphite.
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Diagnostics of Diaphragm Discharge in Water Solutions and its Application for the Nanomaterials Surface Treatment
Dřímalková, Lucie ; Brablec, Antonín (referee) ; Janda,, Mário (referee) ; Krčma, František (advisor)
The exact mechanism of the discharge in liquids ignition is not sufficiently known up to now. Although during the last years was achieved the great progress and overloading which some of them are written in this theoretical part of thesis. This thesis is divided into two experimental parts. When the first part deals with diagnostics of diaphragm discharge in electrolyte solutions and the second part is focused on its use for uncoiling (higher homogenization) of carbon nanotubes in solutions. In experiment 1, three different sized (4 l, 100 ml, 50 ml) diaphragm discharge configurations were used to diagnose diaphragm discharge in electrolyte solutions. Diagnostics is done through current and voltage waveforms with the addition of synchronized ICCD camera images that have been connected to a four-channel oscilloscope. The V-A characteristic can be described by three events occurring in the electrolyte solution with a gradual increase in voltage. Slowly increasing of the voltage in the solution leads first to electrolysis. The next phase is the formation of microbubbles or bubbles, which is characteristic of the curve by a slight decrease in the increase of the current passing between electrodes. The sudden increase in the current flow is characteristic of the last phase, namely the discharge phase. The distance of the electrodes from the diaphragm does not significantly affect the V-A characteristic. The higher diameter of the pin hole, therefore, has a higher voltage, but this does not affect the origin of bubble generation or breakdown. The higher thickness of diaphragm, the higher voltage is needed to the beginning of the bubbles generation, and consequently the discharge breakdown. Comparison of the voltage of the start generation of the bubbles and breakdown for PET diaphragms and diaphragms from the ceramic there was no mark able difference. One of the most important parameters is the conductivity of the electrolyte solution. The lower voltage is needed for the start generation of the bubbles at the higher solution conductivity, and also the discharge generation is observed at a lower breakdown voltage. The second experimental part is focused on the study of the diaphragm discharge effect on carbon nanotubes. A specially designed U-shaped reactor is used to modify carbon nanoparticles. Tap water and aqueous solutions of organic compounds are used as the electrolytic solutions. The discharge is generated by a non-pulsed DC high source with a voltage in the range of 0-2.8 kV supplied to platinum electrodes located in the electrolyte solution. The experimental results have shown that the diaphragm discharge has positive effects on the disintegration of clusters and agglomerates of carbon nanotubes. The primary effect on disintegration is probably the shock waves generated by the discharge. It turned out that it depends on the electrode configuration, where the treatment in anode space has far greater effects than the treatment in cathode half of the reactor. Effects of carbon nanotubes disintegration in solution are long-lasting and the treatment effect is not loosed after several months. There were detected no significant changes in the structure of plasma-treated nanotubes by Infra-red spectroscopy.
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