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Preparation and characterization of enzyme-containing wound dressings
Jurová, Bronislava ; Szotkowski, Martin (referee) ; Skoumalová, Petra (advisor)
This diploma thesis is focused on the study and characterization of nanofiber wound dressings from natural biopolymers. Proteolytic enzymes, specifically collagenase and bromelain, were incorporated into these nanofiber covers. The theoretical part deals with a brief description of skin wounds, their healing and skin covers. There are also methods for preparing these covers and the methods used to characterize them. The practical part deals with the optimization of the preparation of nanofiber coatings based on polyhydroxybutyrate, gelatin, alginate and chitosan. These nanofibers were then enriched with active substances and their gradual release into the model environment was monitored. Finally, their proteolytic activity was determined for these substances.
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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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Preparation of nanoparticles and nanofibers with antimicrobial components
Sosková, Simona ; Veselá, Mária (referee) ; Skoumalová, Petra (advisor)
The presented diploma thesis is focused on the preparation of new materials with antimicrobial effect. Liposomes and nanofibers from polyhydroxybutyrate containing clotrimazole and natural extracts with good antifungal and antioxidant effects were prepared. The theoretical part contains examples and short description of using nanoparticles and nanofibers in cosmetics and medicine and the description of plants which have positive and potential antimycotic effects. Moreover, methods for particles and fibers characterisation were shortly described. In the experimental part, natural water and lipid extracts were prepared and spectrophotometrically characterised for the content of polyphenols, flavonoids and the antioxidant activity. Liposomes and liposomes containtng PHB were prepared from selected extracts and the encapsulation effectivity, shortterm and longterm stability via determination of polyphenols were determined. Prepared particles were characterized with DLS method (size) and zeta- potential (stability). PHB nanofibers functionalised with selected lipid extracts and clotrimazole were prepared via electrospinning and forcespinning, and examined via FLIM and FTIR-ATR methods and spectrophotometry was used for antioxidant activity and release of active substances determination. Antifungal properties of prepared particles, extracts and fibers using the test system Candida glabrata were studied. Finally, cytotoxicity of selected samples was tested with MTT assay using human keratinocytes.
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Polyhydroxyalkanoates and their role in bacterial biofilms
Rucká, Markéta ; Samek,, Ota (referee) ; Obruča, Stanislav (advisor)
This master thesis deals with polyhydroxyalkanoates (PHA) and their role in bacterial biofilms. In the theoretical part the polyhydroxyalkanoates, bacterial biofilm and the relationship between them were reviewed. The experimental part focused on differences in PHA production by planktonic and biofilm cells. In order to study selected topic, bacterial strains of Burkholderia cepacia and Burkholderia sacchari were cultivated using a CDC biofilm reactor. The attention was paid to quantity and especially to the form in which PHA occurs in planktonic and biofilm cells. Results of Raman spectroscopy have shown that PHA exists exclusively in native amorphous form in planktonic bacterial cells. On the other hand, in biofilm PHA occurs also in a partially crystalline form. In addition, the resistance of planktonic and biofilm cells against various stress factors and the effect of osmotic stress on PHA production was tested too. According to the results of the experiment, when the bacteria were exposed to different stress factors (high temperature, low temperature, presence of detergent and so forth) biofilm cells showed a higher stress resistance than planktonic cells. Apart from slowing cell growth and reproduction, increased osmotic pressure in the culture medium also caused decrease of PHA production. In addition, planktonic cells responded to external stimuli more sensitively than biofilm ones.
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Modification of polyurethane by biodegradable poly(hydroxybutyrate)
Kupka, Vojtěch ; Žídek, Jan (referee) ; Vojtová, Lucy (advisor)
Cílem předložené bakalářské práce byla syntéza elastomerního polyuretanu plněného biodegradabilním polyhydroxybutyrátem. Vzorky byly podrobeny analýze morfologie pomocí rastrovací elektronové mikroskopie, mechanické vlastnosti byly testovány jednoosou tahovou zkouškou. Práce se taktéž zabývá předpovědí mechanických vlastností na základě teorií kaučukové elasticity. Bylo zjištěno, že výsledný materiál je částicový kompozit, kde se vzrůstajícím hmotnostním obsahem plniva v materiálu vzrůstá modul pružnosti. Predikce tahových křivek pomocí softwaru SEC.exe ukazovala pro reálný průběh křivky nereálné hodnoty hustot polymerních řetězců. U modelů, které vykazovaly reálné hodnoty hustot, byly naopak hodnoty napětí vyšší, než ve skutečnosti naměřené.
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Utilization of spent wood chips for biotechnological production of PHA
Ladický, Peter ; Sedláček, Petr (referee) ; Obruča, Stanislav (advisor)
The aim of this work was to study the suitability of wood shavings and sawdust as a substrate for microbial production of PHA by bacteria Burkholderia cepacia and Burkholderia sacchari. In the experimental part of the work the most appropriate approach of hydrolysis of wood shaving and sawdust and the effect of polyphenol and furfural concentration on ability to accumulate PHA was studied. Burkholderia sacchari had greater ability to accumulate PHA compared to Burkholderia cepacia. PHB values 87–89 % were achieved when Bulkholderia sacchari was cultivated on medium that contained detoxified hydrolysate of sawdust. Sawdust is therefore a promising substrate for microbial production of PHA in terms of reducing production costs and high content of PHB in biomass.
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Study of the medium influence on the pollutants sorption on microplastics
Hamplová, Marie ; Kučerík, Jiří (referee) ; Mravcová, Ludmila (advisor)
With the increasing production of plastic waste, the pollution of the environment by persistent microplastics is increasing simultaneously. Other environmental pollutants, such as various pharmaceuticals, can be sorbed on microplastics. This increases the toxicity of microplastics and their risk to living organisms. Nowadays, the use of so-called biodegradable polymers, which are supposed to degrade spontaneously, is expanding. However, their degradation can also produce microplastics. Therefore, this bachelor thesis deals with the sorption of drugs onto microplastics from biodegradable polyhydroxybutyrate (PHB) and with the influence of the surrounding aqueous environment on the rate and mechanism of sorption. Sorption of drugs onto microplastics made of synthetic polyethylene terephthalate (PET) was also carried out for comparison. The quantity of drugs was analysed by UPLC-ESI-TQ. Sorption was observed from the drug mixture for antibiotics tetracyclines. The pH value of the surrounding environment influenced the sorption rate of tetracyclines, the highest sorption occurred at acidic pH 4. According to the results of evaluation of sorption by Langmuir and Freundlich adsorption isotherms, PHB microplastics have a higher sorption capacity than PET microplastics. Therefore, PHB microplastics could be a risk for living organisms.
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Encapsulation of active substances into nanofibers and possibilities of their application
Procházková, Lucie ; Pernicová, Iva (referee) ; Skoumalová, Petra (advisor)
The master thesis was based on the optimization of the production of nanofiber covers and to gaine the product for subsequent functional use. The production of nanofiber covers was made by electrospinning and forcespinning from selected materials. Polyhydroxybutyrate, gelatin, chitosan and alginate were used as starting materials. After successful optimization, these materials were enriched with active ingredients ampicillin and ibuprofen for the functionalized use of covers for more effective wound healing. The theoretical part was focused on the issue of skin, healing processes, types of wounds and nanofibers, the characterization of selected starting materials for the formation of nanofibers was also mentioned. The practical part was based on the lengthy optimization of the preparation of fiber covers and later enriched with active ingredients. Furthermore, combined covers made of different materials with contents of both active ingredients were designed. This was followed by the characterization of all prepared covers from the point of view of stability in the short and long term. The gradual release of active ingredients was determined spectrophotometrically and by hifh performance liquid chromatography. It was also important to determine the antimicrobial activity of selected active substances. At the end of all testing, combined coatings containing both active ingredients were used for safety testing with human keratinocyte cells (HaCaT). Safety testing was based on determining the viability of human cells using the MTT test, to verify the LDH test. A scratch test was also performed, a wound healing test after the application of devised combined covers.
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Utilization of physico-chemical and spectroscopical methods in study on microbial extremofiles
Dobešová, Kateřina ; Enev, Vojtěch (referee) ; Sedláček, Petr (advisor)
The aim of this bachelor thesis is the determination od polyhydroxyalkanoates in the cells of extremophille bacterie, preformed by several methods. The chosen bacterium was Schlegelella thermodepolymerans DSM 15 344 Thermogravimetric analysis, infrared spectroscopy with Fourier transformation, and Raman spectroscopy were used in this thesis. The TGA method was used for the determination of extracellular and intracellular water in samples. The FTIR method was used in order to observe the amount od PHA, but aslo to observe the physical properties of the polymer in the sample, specifically to determine either amorphous or crystalline featuers of this polymer. Raman spectroscopy was used in oder to confirm the presence of PHA in the samples. All results e´were compared eith the results accomplished by gas chromatografy.
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Study of the interaction of selected pharmaceuticals with microplastics
Ioani, Aldina ; Řezáčová, Veronika (referee) ; Doležalová Weissmannová, Helena (advisor)
The bachelor thesis aims to study the interaction of selected pharmaceuticals from the group of analgesics and antibiotics with microplastics. The analysis was conducted in the mid-infrared region, which is used to identify chemical structure of the substances. Fourier transform infrared spectroscopy was used as an analytical method. The samples were subjected to technique attenuated total reflection. Ibuprofen and trimethoprim were chosen as analytes due to their common usage in the population. Sorption of pharmaceuticals was undergoing on polyhydroxybutyrate, which is a kind of biodegradable plastics and polyethene terephthalate, which is synthetic. Measured results showed that for the study of interaction with pharmaceuticals, it is more suitable to use biodegradable plastic polyhydroxybutyrate than polyethene terephthalate because of its better results. Infrared spectra output is very influenced by a selection of a solvent.
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