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Biocrystallization as a new concept in development of piezoelectronics for use in biomaterials
Dobešová, Kateřina ; Plichta, Tomáš (referee) ; Sedláček, Petr (advisor)
This thesis focuses on biofilament methods and biocrystallization of bacterial cells producing polyhydroxyalkanoates, specifically the bacterium Cupriavidus necator H16. The main techniques used for cell orientation include spin-coating and acoustic waves. Biocrystallization was induced by temperature, osmotic, and acid stress. The degree of cell orientation was monitored by an atomic force microscope, and crystallization was determined by FTIR. Among other parameters, the thickness of the layers was evaluated using a profilometer. The amount of PHA in the bacterial cells was determined by gas chromatography. The preparation of crystalline ordered layers of biomaterials represents a route to piezoelectric biomaterials.
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Polymeric blends of thermoplastic starch with other biodegradable polymers
Nováková, Pavlína ; Petruš, Josef (referee) ; Kučera, František (advisor)
The master's thesis deals with the preparation and properties of biodegradable polymer blends of thermoplastic starch (TPS) with polyhydroxybutyrate (PHB) with emphasis on the compatibilization of the immiscible polymer blend. The theoretical part focuses on the characterization of thermoplastic starch, thermodynamics of polymer blends, possible ways of compatibilization of polymer blends and a summary of the current knowledge on polymer blends of thermoplastic starch with biodegradable polymers. The experimental part deals with the modification of starch and PHB with maleic anhydride in order to compatibilize the polymer blend. The binding of the monomer to the polymers was confirmed by infrared spectroscopy and acid-base titration. Subsequently, using the prepared modified polymers, TPS/PHB blends were prepared and the effect of compatibilization and blend composition on the properties was evaluated. Differential scanning calorimetry (DSC) was used to observe the melting and crystallization process, the changes in molecular weight of the polymers were assessed by determining the melt flow index. Tensile strength, elongation at break and the effect of ageing on the mentioned mechanical properties were determined by tensile testing, morphology was observed by electron microscopy (SEM).
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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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Preparation and characterization of nanoparticles with encapsulated antimicrobial component
Dohnalová, Marie ; Strečanská, Paulína (referee) ; Márová, Ivana (advisor)
This bachelor thesis is focused on the preparation and characterization of particles with encapsulated natural substances with antioxidant and antimicrobial properties. Selected antioxidants were anthocyanins, eugenol and vitamin E, as one of the most powerful antioxidants. At the same time, antimicrobial agents were chosen, in this case eugenol. In the theoretical part, individual active substances, materials for the preparation of particles, methods of their preparation were characterized and the principle of determining the safety of used materials. In the experimental part, selected active substances were characterized and the preparation of particles was optimized. Two types of liposomes were prepared, for which the encapsulation efficiency, size and stability were determined. Also prepared aqueous extracts of blueberries and blackberries were prepared, for which total anthocyanin content has been determined. The extracted substances from the plant material were encapsulated into alginate particles. Furthermore, the gradual release of active substances from particles into model environments using a spectrophotometer was determined. During the gradual release of active substances from the particles, the antioxidant effect was also monitored, in which it was possible to observe a decrease with the passage of time. The antimicrobial effect of active substances and liposomes against model microorganisms Escherichia coli and Micrococcus luteus was also tested. Cytotoxicity tests on human keratinocytes were performed on selected active substances, extracts and prepared nanoparticles, where it was proven that the selected materials are non-cytotoxic. Finally, a proposal for a possible application form in the form of an alginate hydrogel was created.
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Preparation of skin masks with the anti-acne effect
Geierová, Kateřina ; Bendová, Agáta (referee) ; Márová, Ivana (advisor)
This bachelor thesis was focused on the preparation and characterization of nanofibers containing active agents with the aim of using these materials in anti-acne facial masks. The nanofibers were prepared from polyhydroxybutyrate solution and gelatin solution by electrospinning method. Among the active ingredients, eugenol and piperine were used to functionalize the nanofibers. In the theoretical part, the types of face masks, the individual active ingredients, the materials for the preparation of nanofibers, their preparation methods and their characterization were characterized. In the practical part, the antioxidant and antimicrobial activity of the active ingredients themselves was investigated. Subsequently, the release rate of selected active ingredients from the prepared nanofibers was monitored in three model environments. The antimicrobial activity of the nanofibers was monitored on three microorganisms, namely Staphylococcus epidermidis, Cutibacterium acnes and Escherichia coli.
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Proposal of systems for targeted transoprt of compounds of sweet taste
Demová, Radoslava ; Hlaváček, Viliam (referee) ; Márová, Ivana (advisor)
This bachelor thesis is focused on possibility of immobilization sweeteners on polymeric system. In the theoretical part information about sweeteners, carrier systems and the possibility of immobilization technique were reviewed. In the experimental part preparation of nanoparticles of polyhydroxybutyrate (PHB) was optimized. The PHB particles were characterized by dynamic light scattering (DLS). PHB particles were activated by plasma treatment. The surface morphology was studied using infrared and Raman spectroscopy. Immobilization of following substances with a sweet taste was tested: glucose, fructose, sucrose, mannitol, sorbitol and erythritol. From these compounds only sucrose was immobilized on PHB particles successfully. Prepared immobilized particles were exposed to the artificial stomach juice, intestinal juices and bile acids and amount of released sucrose was monitored. Finally, long-term stability prepared particles was measured and also the amount of sucrose released was determined.
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Preparation of nanoparticles and nanofibers with antimicrobial components
Kubišová, Veronika ; Slaninová, Eva (referee) ; Skoumalová, Petra (advisor)
This thesis addresses the problem of inadequate current wound therapy and presents a solution in the form of nanomaterial-based wound dressings (coverings). Specifically, it focuses on the development and characterization of various nanofibrous materials with integrated liposome particles that could serve as a source of therapeutic agents and be useful specifically in the field of a wound therapy. The review in the introductory part of the thesis first focused on the mentioned problem, which describes the shortcomings of existing conventional dressing materials. A description of human skin from an anatomical and functional point of view was not omitted, nor was the wound healing process itself. Different types of covering materials were also presented. However, a large part was focused on nanomaterials and their use in the field of the skin wound therapy. The nanomaterials mentioned were mainly liposome particles and nanofibres, as the experimental part of the work was focused on these structures. The description dealt with the characterization of these structures as well as the biopolymers used for their preparation. However, the aim was to prepare nanofibers with liposome content, so the method of forming such systems was described. Various therapeutic agents are also an integral part of the covering materials, especially those that suppress the development of infection and reduce wound pain; therefore, the search focused on the antibiotic ampicillin and the analgesic ibuprofen. The experimental part was devoted to the actual production of nanofibers with liposomes and also to the qualitative demonstration of the presence of liposomes in electrostatically prepared nanofibers. The selected polymeric components of these systems were polyhydroxybutyrate (PHB) and gelatine. However, the nanofibers and liposome particles (as well as combined liposomes with PHB) were first prepared in their own form and characterized mainly in terms of the gradual release of the drug substances. The results obtained were then compared with those of therapeutic drug delivery via combined nanofibrous structures with liposomes. For these combined structures, the aim was to achieve synergy in drug delivery between these systems. The aim of this work was to create a biomaterial covering with the controlled drug release. The drugs contained in these materials were the aforementioned ampicillin and ibuprofen. An important part of the work was then to determine the safety of the prepared materials which were tested for cytotoxicity, where the tests performed were MTT test and LDH test. And the actual wound healing ability of the nanofibers was then monitored in a scratch test or a "wound healing" test. At the end of the paper, recommendations for future work on this topic are given.
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Influence of biodegradable polymers microplastics on properties of soil organic matter
Denková, Pavla ; Řezáčová, Veronika (referee) ; Kučerík, Jiří (advisor)
The microplastics contamination is an issue observed in all environmental compartments. Currently, the trend is to replace the synthetic polymers by biodegradable ones, which are expected to degrade faster due to omnipresent microorganisms. However, this assumption may not correspond with the reality. In nature, biodegradation can last long or not whatsoever and the microparticles of biodegradable polymers can remain in soil the same way as particles of synthetic polymers. The effect of microparticles of biodegradable polymers on soil´s biota has already been studied, but the effect on the properties of soil and water in the soil has not been elucidated yet. This thesis deals with the influence of microparticles of biodegradable plastics on the physico-chemical properties of soil organic matter, especially on the behavior of water in the soil system under arid and semiarid conditions. Microparticles of poly(R-3-hydroxybutyrate), abbreviated as P3HB, were used as a model biodegradable polymer and were introduced into the soil in various concentrations. To determine the evaporation enthalpy of water and stability of water bridges that affect (stabilize) the physical and chemical structure of soil organic matter Differential Scanning Calorimetry (DSC) was used. Simultaneously was also measured their effect on water holding capacity of soil organic matter. Under experimental conditions, P3HB microplastics were found to reduce evaporation enthalpy, which facilitates drying of soil organic matter. Nevertheless, their effect on final water holding capacity was relatively low. Besides, we observed a negative effect on the stability of water molecule bridge that connect segments of soil´s organic components. Thus, P3HB microparticles in soil could pose a risk due to their influence on the water retention mechanism in soil under arid and semiarid conditions.
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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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Utilization of physicochemical and spectroscopic techniques in study of stress-response of cyanobacteria
Skoryk, Maksym ; Šedrlová, Zuzana (referee) ; Sedláček, Petr (advisor)
Tato bakalářská práce je soustředěná na zkoumaní cyanobakterií, vystavených hyper a hyposmotickým stresům. Na základě literární rešerše byly navžené vhodné analytické metody určené ke studiu dvou modelových organismu - Synechocystis sp. PCC 6803 a Synechocystis sp. salina Wislouch CCALA 192. Celkem čtyři metody byly použité k charakterizaci osmoticky zatížených bakterií. Průtoková cytometrie byla použita k vyhodnocení viability. Fluorescenční sonda SYTOX Blue poskytla důvěryhodnou informaci ohledně viability bakterií. Propidium jodid naopak poskytl nezřetelnou informaci. Optické vlastnosti cyanobakterií byly prozkoumané pomoci UV-VIS absorpčních a turbidimetrických měření. Termogravimetrická analýza byla použita pro mapování změn obsahu vody v osmoticky stresovaných buňkách. Tato metoda ukazala. že PHB-positivní Synechocystis sp. PCC 6803 jsou pravděpodobně vice odolné vůči hyperosmotickým stresům než PHB-negativní. Plynová chromatografie byla použita ke kvantifikaci vnitrobuněčného PHB, který činil přibližně 1-2 % suché hmoty PHB-positivních buněk.
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