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Synthesis of spherical gold nanoparticles for biomedical applications
Gablech, Evelína ; Adam, Vojtěch (referee) ; Drbohlavová, Jana (advisor)
Tato práce se zabývá syntézou sférických zlatých nanočástic pro biomedicínské aplikace. Zlaté nanočástice byly syntetizovány ekologicky nezávadnými metodami, kterým je věnována i značná část rešerše. Cílem bylo nasyntetizovat stabilní koloidní zlaté nanočástice vhodné pro různé biomedicínké aplikace zejména, pro in vivo a in vitro zobrazovací metody, kterých přehled je take obsažen v teoretické části práce. Také byl proveden test cytotoxicity, jelikož částice mají být použitelné pro in vivo aplikace. Částice byly dale charakterizovány metodami SEM, DLS a UV-VIS.
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Ion-selective membrane with antimicrobial effect
Gablech, Evelína ; Farka,, Zdeněk (referee) ; Soukupová,, Jan (referee) ; Drbohlavová, Jana (advisor)
Tato práce se zabývá zlepšením antimikrobiálních vlastností komerčních polymerních membrán používaných v mlékárenském průmyslu pro filtraci mléčných výrobků jako je syrovátka. Tyto membrány trpí kontaminací způsobenou mikroorganismy přítomnými v mléce během procesu filtrace. Toto biologické znečištění ovlivňuje životnost membrány i samotný filtrační proces. Pro ochranu membrány před vytvořením biofilmu z mikroorganismů byly tyto membrány modifikovány pomocí plazmových metod tak, aby došlo k funkcionalizaci aminovými skupinami pro následnou imobilizaci nanočástic stříbra a selenu. Antimikrobiální aktivita obou typů nanočástic a membrán s imobilizovanými nanočásticemi byla stanovena proti buněčným kmenům Staphylococcus aureus a Escherichia coli.
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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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Functionalization of gold nanoparticles for imaging
Jakubechová, Jana ; Pekárková, Jana (referee) ; Drbohlavová, Jana (advisor)
The diploma thesis deals with synthesis of gold nanoparticles and their surface functionalization suitable for in vitro imaging. In this view there are requirements for optical properties such as stability, monodispersity and no presence of cytotocxicity. In order to fulfill these demands the synthesis by Turkevich method with surface modification by glutathione and polytethylenglycol was performed. Analytical methods such as DLS, SEM and Zeta potential measurement were utilized to characterize the physical and chemical properties of synthesized gold nanoparticles. Finally, MTT assay was performed to evaluate toxicity of gold nanoparticles using HEK 293 cell line.
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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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Biocomposite material for 3D print in the field of regenerative medicine
Chaloupková, Kateřina ; Obruča, Stanislav (referee) ; Přikryl, Radek (advisor)
The presented thesis deals with preparation of material for use in regenerative medicine based on poly(3-hydroxybutyrate) and its characterization. In addition to poly (3-hydroxybutyrate), there were used other materials lactic acid (PLA), tricalcium phosphate (TCP) and two types of plasticizers Citroflex®B-6 (CB6) and Syncroflex3114 (S3114). These materials were selected based on their biocompatibility and, in the case of TCP, also bioactivity. TCP allows new bone to grow on the surface of the scaffold. PLA was used to improve the mechanical properties of the material. Both plasticizers have been used to improve the processability of the material. Theoretical part of this work contains a literature review describing basic information about used materials. Aim of the experimental part is to prepare the material, characterization of properties and determination of printability on a 3D printer. The material is examined for thermal properties by thermogravimetric analysis and differential scanning calorimetry. This work also deals with the matter of 3D printing, especially FDM technology. It has been found that materials containing the syncroflex plasticizer are better processed and therefore printed on a 3D printer. The printability tests performed are temperature towers and filling studies. Printed samples were subjected to mechanical tests of tensile and bending tests. Experiments of cytotoxicity and biocompatibility of the material were also performed. Within the work, TCP particles were characterized using a particle size analyzer. The average TCP particle size is 10,76 µm. Using SEM-EDX, the distribution of TCP in sample filaments was subsequently observed, where it was found that by mixing TCP particles with the remaining components of materials, TCP particles agglomerate into formations up to 20 µm in size. Roughness of materials was determined by confocal microscopy. Cytotoxicity was also tested in the extracts of samples on mouse fibroblasts. Cytotoxicity was determined by metabolic activity assay and light microscopy. The metabolic activity test proved the biocompatibility of the observed materials; therefore, it was possible to perform cell proliferation and biocompatibility tests directly on the samples. Assays were performed using human mesenchymal stem cells. DNA quantification was used to determine cell proliferation. Shape of cells was subsequently observed by confocal microscopy. Tests confirmed growth of cells and their appropriate shape. Stem cell differentiation into bone was performed by measuring alkaline phosphatase activity.
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New possibilities in wound healing
Nováková, Laura ; Szotkowski, Martin (referee) ; Skoumalová, Petra (advisor)
The diploma thesis is focused on the study of fibrous wound dressings prepared by electrospinning method from natural biopolymers. Three active ingredients were added to the dressings: ampicillin, ibuprofen and collagenase, which are responsible for relieving pain, reducing the risk of infection and selectively removing necrotic tissue in the wound. The theoretical part describes the therapeutic dressings currently available on the market and the most common methods of nanofiber production. The experimental part evaluates the optimization of the preparation of gelatin, alginate and chitosan fibrous wound dressings, which were subsequently enriched with active substances and their gradual release into the model environment was determined spectrophotometrically. Antimicrobial effects against E.coli and S. epidermidis strains andantifungal activity against C. glabrata yeast were monitored. Finally, two cytotoxicity tests on the human keratinocyte cell line HaCaT confirmed the safety of the prepared products, which can serve as bioactive skin dressings in the future.
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Analysis of active substances and biological effects of some non-tradizional cereals
Pecháček, Michal ; Vysoká, Marie (referee) ; Márová, Ivana (advisor)
This thesis deals with the characterization of active substances and biological effects of non-traditional cereals. Basic substances of these non-traditional cereals such as carbohydrates or proteins are characterized in this work. Active compounds such as antioxidants, polyphenols or -glucans are also characterized. These substances were mainly analysed by spectrophotometric methods. The theoretical part describes cereals in general. It focuses on grain morphology, cereal products and mostly on chemical composition. The summary of substances appearing in the cereal grain such as carbohydrates, proteins, vitamins, minerals etc. was processed in this part. In the practical part, the samples of the non-traditional cereals such as amaranth, sorghum, millet, kamut, buckwheat, quinoa, Job’s tears and teff were analysed. Amaranth, millet and buckwheat were also analysed in the form of flakes. Teff was analysed only in the form of flakes. Sorghum and quinoa were analysed also in their coloured variations. Firstly, the water extracts were prepared, then they were used for determination of antioxidant activity, the content of polyphenols compounds and flavonoids. The hydrolysed samples were used for determination of the carbohydrates. For other basic analysis, samples were used in the form of powder. Best results were measured for quinoa, amaranth and buckwheat. Besides basic analysis, the content of -glucans was measured. However, the content of -glucans in these non-traditional cereals was very low. Selected cereals were tested for cytotoxicity on human cells. Cytotoxicity was evaluated by using the MTT cytotoxicity test on human keratinocytes HaCaT and human caucasian colon adenocarcinoma CaCO-2. The sensory analysis was carried out in the last part of the thesis. There were tested 6 samples. From these samples were made muffins that were analysed and sensorically evaluated.
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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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Stability of PHA-based particles and fibres in different environments
Tarageľ, Matej ; Skoumalová, Petra (referee) ; Márová, Ivana (advisor)
The aim of this diploma thesis is the preparation of liposome nanoparticles enriched with PHA and PHA nanofibers. The nanostructures served to encapsulate extracts of lipophilic and hydrophilic nature. The characterization of the properties of nanostructures such as polydispersity, size, colloidal stability, long-term stability after exposure to various environments such as seawater, water from the Brno dam and tap water, and finally the cytotoxicity of fibers with extracts was addressed. The theoretical part is focused on different types of water, human skin, coffee and subsequently carotenoids. It continues by describing of possibilities of extraction and preparation of lipophilic and hydrophilic extracts and possibilities of their determination is discussed. Finally, it describes the possibilities of preparation and characterization of PHA based nanomaterials. The practical part deals with the preparation of liposome particles and fibers enriched with PHA with encapsulated extracts, their characterization, and their subsequent exposure to various environments. Monitoring of their long-term stability was carried out, but the release of the encapsulated extracts into the environment to which the nanoparticles and nanofibers were exposed was also measured. Finally, the interaction of nanofibers with live HaCaT cells was monitored, and cytotoxicity assays determined the viability of the cells after interaction with the nanofibers.
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