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Characterization and application of microwave plasma on wound healing
Truchlá, Darina ; Němcová, Andrea (referee) ; Krčma, František (advisor)
Non-thermal plasma has a lot of ways for using in nowadays medicine. It presents many useful actions like charged particles, UV light, electric field, radicals, excited atoms and molecules. That complicated chemistry directs to uncountable synergistic interaction between cold plasma and biological systems, involve cells and tissues. This thesis is about effects of cold plasma to wound healing. Two different microwave plasma systems were used for the presented study. The first one was argon plasma torch generated by surface wave using the quartz capillary, the second one was plasma torch with reverse vortex argon flow. Diagnostics of plasma jet by optical emission spectroscopy shown the presence of active particles, which are responsible for a lot of impact of plasma treatment. Concentrations of active particles generated by plasma are dependent on conditions of plasma generation like power of generator and gas flow. For visual evidence of effects on skin caused by active particles was created simulation of skin tissue. Interaction between plasma jet and artificial skin tissue shown that UV light and temperature are not responsible for all observed effects which are noticed after plasma treatment. Some part of experiments was realized in collaboration with Medical University of Sofia in Bulgaria. The theory of positive effect to wound healing was supported by experiments based on treating artificially created wounds on laboratory mice by cold plasma. It was proved, that process of wound healing is significantly shorter after using plasma treatment in comparison with normal wound healing. Plasma treating of wound for 10 seconds in two consequent days seems like more effectively than application of plasma only one day. This Thesis was carried out as a part of international project PLASMABORDER that was supported by European commission under cohesion funds; programme INTEREG SK-CZ under contract No. 304011P709.
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Use of plasma jet for wound healing
Dvořáková, Eva ; Márová, Ivana (referee) ; Skoumalová, Petra (advisor)
This diploma thesis was focused on the possibility of using a plasma nozzle to accelerate the wound healing process. The benefits of using low-temperature plasma in medicine or biomedical applications are known from many studies, and low-temperature plasma is already used to sterilize medical devices, materials or surgical instruments. Some studies also report a high potential of usinh plasma nozzle in the treatment of skin wounds. In the experimental part of this work, an in vitro wound healing test was performed using two different low-temperature plasma sources. Source No. 1 was a surface wave microwave discharge and source No. 2 was a torch microwave discharge. An in vitro scratch healing test was performed on a monolayer of HaCaT keratinocytes and testing was performed using various parameters. The influence of the plasma treatment time was monitored, as well as the influence of the plasma discharge power and also the influence of the argon working gas flow. Especially when using a torch microwave discharge, faster wound healing was recorded at most of the parameters used compared to the control. Thus, it can be said that this source appears to be potentially suitable for faster wound healing. Furthermore, in the work using the MTT cytotoxicity test, the viability of skin cells after their plasmination was also monitored using the same conditions as in the in vitro wound healing test. When performed in the standard MTT assay, none of the settings or sources used showed any cytotoxic effects on keratinocytes. LDH cytotoxicity tests were also performed concurrently to verify the accuracy of the MTT assays. The results of both tests agreed and the use of low-temperature plasma in skin treatment can be considered as safe. Overall, the results show that the plasma nozzle can find use in medicine in the healing of skin wounds and chronic defects as a potentially fast, inexpensive and effective method.
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Characterization and application of microwave plasma on wound healing
Smejkalová, Kateřina ; Skoumalová, Petra (referee) ; Krčma, František (advisor)
The aim of the Master thesis is the investigation of the influence of microwave discharge for skin wound healing. Microwave discharge used for this work was argon microwave plasma generated by the surface wave and direct vortex torch. The theoretical part is focused on basic information about plasma and processes that occur in plasma discharge under specific conditions. Plasma generates various active particles such as hydroxyl radicals, nitric oxide radicals, excited nitrogen molecules, atomic nitrogen, argon and oxygen. All of these particles together with plasma generated photons are usable in biomedical applications and summary of them is shown in the theoretical part. The experimental part is focused on the comparison of torch discharge and microwave plasma generated surface wave in skin wound healing. The model wounds on laboratory mousses were treated by plasma and wound healing was examined during 3 weeks after the plasma treatment. Both plasma systems showed healing acceleration. Application of torch discharge was proved to be the most effective method in the healing of skin defects. Additionally, determination of active particles was taken by optical emission spectroscopy. Based on these measurements, plasma parameters were determined: electron temparutare, rotational and vibrational temperatures. To determine role of different plasma active species, the treatment of indigo coloured artificial skin model was treated under various conditions by both plasma systems. Results show that the direct interaction between plasma particles is the main effect, role of radiation, only, is more or less negligible. Finally, the plasma vortex system was visualized using fast camera at selected powers and gas flows.
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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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Study of plasma interaction with bacteria for wound healing
Šrámková, Sarah ; Krčma, František (referee) ; Kozáková, Zdenka (advisor)
This thesis focuses on the study of the interaction of plasma with bacteria to enhance the promotion of wound healing. Firstly, the wound healing process is described, followed by low temperature plasma, its effect on bacterial cells and its use in biomedicine. The experimental part deals with the application of low-temperature plasma generated by microwave jet on selected microorganisms and the influence of experimental conditions on the antimicrobial effect of the plasma. One representative of the microorganisms occurring in the wounds was selected as representatives of Gram-positive bacteria and Gram-negative bacteria, namely Staphylococcus epidermidis and Escherichia coli. Using the results obtained, the antimicrobial effect of plasma generated by microwave jet in argon was confirmed. The degree of antimicrobial effect is related to the concentration of the microorganisms, the time of treatment and whether the bacteria are Gram-positive or Gram-negative.
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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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Development of antimicrobial wound healing nanostructure biomaterials
Hanák, Jan ; Wikarská, Monika (referee) ; Márová, Ivana (advisor)
The bachelor thesis is focused on optimizing the production of nanoparticles with a bioactive component - especially on enzymes and antibiotics encapsulated into lipozomes. Nanoparticles with active substances could be used in the field of pharmacy or cosmetology. In the theoretical part the composition of the skin, skin injuries, and wound covers are described. The materials used for the preparation of nanoparticles and nanofibers are characterized as well as processes to produce such nanomaterials. Enzymes, antimicrobial drugs, and methods for determining antimicrobial activity are mentioned in this section, too. The practical part deals with the characterization and optimization of the formation of lipozome particles with encapsulated enzymes. The formed particles were divided into two groups. The first type of particles were pure lipozomes, while the second ones enriched by polyhydroxybutyrate to increase the encapsulation efficiency and improve the stability of the particles. The enzymes bromelain and lysozyme were selected for the highest encapsulation efficiency. The encapsulation of these two enzymes took place in two ways - by thin film hydration and by sonication method. Both methods showed high encapsulation efficiency, but the sonication method proved to be unsuitable due to enzyme denaturation. The proteolytic activity of the released enzymes from the particles was measured. The stability of the particles was monitored in a physiological and aquatic environment by measuring the zeta potential for up to 28 days. Moreover, the enzymes, ampicillin, and particles created by sonication were subsequently subjected to antimicrobial tests by using two test bacterial strains. Finally, alginate particles with encapsulated enzymes were also formed, then the encapsulation efficiency and proteolytic activity of the enzyme were measured. In this study also other methods of encapsulation of active substances are outlined, which could be possibly used to produce nanomaterials suitable for wound healing.
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Modification of hydrogel wound dressings by biological active agents
Štrublová, Veronika ; Říhová, Martina (referee) ; Vojtová, Lucy (advisor)
Cílem práce je navrhnout a připravit cenově výhodný hydrogelový kryt ran, který bude chránit ránu před mikrobiální infekcí a urychlovat proces hojení. Teoretická část shrnuje jak přírodní, tak syntetické polymery, antibakteriální a pro-hojivá aditiva, která jsou široce používána v biomedicínských aplikacích. Obsahuje také krátký přehled jednotlivých fází hojení ran. Experimentální část předložené práce se zabývá přípravou a charakterizací hydrogelových krytů ran s porézní strukturou složených jak ze syntetických, tak přírodních polymerů zahrnujících přírodní polysacharid Gum Karaya (GK), polyvinylalkohol a polyethylenglykol. Připravené lyofilizované hydrogelové kryty ran byly obohaceny biologicky aktivními látkami – antibakteriálními nanočásticemi selenu a fibroblastovým růstovým faktorem FGF2, který představuje protein pro hojení ran. Pro charakterizaci lyofilizovaných hydrogelových krytů ran byla použita řada fyzikálně-chemických technik. Metoda zeslabeného úplného odrazu infračervené spektroskopie s Fourierovou transformací (ATR-FTIR) byla použita ke stanovení změn v chemické struktuře, biomechanické vlastnosti za fyziologických podmínek byly vyhodnoceny pomocí tahové zkoušky za použití dynamicko-mechanické analýzy, morfologie byla charakterizována skenovací elektronovou mikroskopií (SEM) a botnací vlastnosti připravených vzorků byly zjištěny vhodnou gravimetrickou metodou. K vyhodnocení antimikrobiálních vlastností GK byla použita jak mikrodiluční bujónová metoda, tak difúzní agarová disková metoda. Cytotoxicita bylo kvantifikována na základě A) počítání počtu živých a mrtvých buněk v extraktech z GK a B) měřením buněčné proliferace pomocí resazurinové eseje. In vivo testování proběhlo ve spolupráci s Výzkumným ústavem veterinárního lékařství na praseti bílém jako animálním modelu. Z výsledků vyplynulo, že vzorky obsahující glycerol vykazují vynikající mechanické vlastnosti jak za suchého, tak za mokrého stavu. Výsledky antibakteriálních testů ukázaly pozitivní synergický efekt GK a selenových nanočástic na inhibici růstu všech testovaných G+ kmenů. In vivo a in vitro testy prokázaly zvýšenou hojivou schopnost připravených materiálů bez způsobení cytotoxických reakcí. Nové porézní a velmi stabilní hydrogelové kryty ran jsou nadějnými materiály, které mohou být využity v biomedicínských aplikacích, především v oblastech vlhkého hojení ran.
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Chronic wound care - shin ulcer
Kümmelová, Tereza ; Hromádková, Jaroslava (advisor) ; Nikodemová, Hana (referee)
The bachelor thesis Chronic wound care - shin ulcer deals with the issue of chronic wounds focusing on tibial ulcers. The aim of the thesis was: To collect data on treatment options for and to study the available resources. To obtain data from the database "Care of the patient with a non-healing wound" at the University Hospital in Motol. These data were then assessed and statistically processed. To calculate a qualified estimate of the burden of nursing staff related to the care of patients with shin ulcer in internal medicine units, surgical units and in general in the University Hospital in Motol. The theoretical part of the thesis gives a basic overview of the problem. The empirical part of the thesis presents the results of the own investigation and their comparison with the results of previously conducted studies. The method of the research investigation was secondary data analysis from the database "Care of a patient with a non-healing wound" at the University Hospital in Motol. The research investigation included 164 patients with shin ulcer hospitalized in the period from January 1, 2022 to June 31, 2022 at the University Hospital in Motol. Results: It was found that more males were hospitalized with shin ulcer than females in Motol University Hospital during the period. More patients with...
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Changes in the manner of dressing of surgical wounds in the last 30 years
SPILKOVÁ, Eliška
Surgical wound care - dressing is a set of activities that favorably affect the healing process. The course of dressing under aseptic conditions is important to avoid infection in the wound and unpleasant complications. The aim of this diploma thesis is to analyze the changes in the way surgical wounds are used, especially in the last three decades. In connection with the analysis of the text, a research question was asked: How has the method of dressing surgical wounds changed in the last 30 years? We set 5 hypotheses. H1: nurses who have been in practice for more than 10 years prefer sterile devices stored in containers to wound dressings over nurses who have been in practice for less than 10 years. H2: Nurses who have been in practice for more than 10 years prefer the no-dose system more often than nurses who have been in practice for less than 10 years. H3: Nurses who have been in practice for more than 10 years have seen more changes in the method of dressing in the last 30 years than nurses who have been in practice for less than 10 years. H4: Nurses who have been in practice for more than 10 years claim that there have been no changes in the disinfectant and treatment solutions used for dressings during their practice. H5: From the point of view of nurses who have been in practice for more than 10 years, wound dressings are performed more aseptically now than 30 years ago. The empirical part of this diploma thesis was processed on the basis of qualitative and quantitative research. Data collection was performed through a questionnaire and a semi-structured interview. After saturating the answers, a total of 10 interviews and 272 questionnaires were included in the empirical part of this work. The results of the research show that the main changes that have occurred in the last three decades include changes in medical and disinfectant solutions, wound dressings, sterilization packaging, sterile material handling system, wound dressings used and, last but not least, wound healing .
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