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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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Role of matrix metalloproteinases in regulation of late embryonic healing process
Kikinderová, Paulína ; Šindelka, Radek (advisor) ; Procházka, Jan (referee)
Cutaneous wound healing could be distinguished into two main types: embryonic and adult. Embryonic healing in contrast to adult is faster, scar-less and consists of early, middle, and late phases. Actin ring is formed during the early phase and its cables pull the edges of the wound towards apposite sites during the following middle phase. De novo expression of healing specific genes is initiated also during middle phase. However, process of the wound healing continues under the closed wound in the late phase which has been poorly described. Adult wound healing is more complex, longer, and is divided into 4 phases: haemostasis, inflammation, proliferation, and remodelling phase. Adult wound healing might end with the scar. Pivotal role in the wound healing is given to matrix metalloproteinases (MMPs). These remodelling enzymes are important for releasing cytokines, inducing apoptosis, and degradation of extracellular matrix. Our laboratory performed temporal RNA-sequencing of the healing tissue using tailbud stage and swimming tadpole embryos. Results showed predominant expression of four mmps: mmp1.L, mmp7.S, mmp8.S, and mmp9.L. Injury or amputation caused the upregulation and their expression level peaked at 3-6 hours post injury which corresponds with late phase of healing. Thus, I focused on...
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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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Role of skin stem cells in aging and regeneration
Hlaváčková, Tereza ; Nešporová, Kristina (advisor) ; Procházka, Jan (referee)
Skin is a natural and effective barrier of the body against the adverse effects of the external environment. Which, however, requires its constant regeneration and, in the event of damage, repair. Stem cells stored in the epidermis, hair follicles and dermis contribute to this. As with other stem cells, these are characterized by so-called stemness, ie the ability to self-renew and differentiate into other cell types, thus providing a source of cells for skin renewal. During aging, due to internal and external factors (mainly due to oxidative stress and DNA damage), the integrity and functionality of the skin barrier are lost. This process is related, among other things, to a reduction in the number and function of skin stem cells. Today several therapeutic approaches are being developed that use stem cells, but at the same time, it is clear that their origin also significantly affects their use. Therefore is necessary to have a good understanding of the specific properties of the function of skin stem cells to modulate the properties of the skin. This work aims to create a review of scientific literature, which covers the topic of skin stem cells, their role in the processes of regeneration and repair and their role in aging. The work will also address the issue of the skin or other types of stem...
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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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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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The role of nitric oxide during embryonic wound healing and regeneration
Abaffy, Pavel ; Šindelka, Radek (advisor) ; Brábek, Jan (referee) ; Krylov, Vladimír (referee)
The study of the mechanisms that control wound healing is an attention-drawing area within the fields of biology and medicine. Wound healing can be usually defined as two basic types. The first type is adult wound healing, which is slow and results in the scar formation. The second type is referred to as embryonic wound healing, which is in contrast fast and scarless. Wound healing is a complicated process that includes many steps, which are regulated by various types of molecules. One of these important molecules is nitric oxide (NO). Its function is usually connected with the regulation of inflammation and angiogenesis during adult wound healing. However, there is currently no information on its role during embryonic wound healing, where the immune and vascular systems are not yet developed. In this work, we explore and describe the role of the NO during the healing of the early embryos. The highest concentration of the NO post wounding is produced during the first 30 minutes after injury. This applies to all developmental stages, from the blastula stage all the way to the swimming tadpole stage. The main role of the NO during embryonic wound healing is the regulation of the gene expression that is connected with the stress response and the regulation of cellular metabolism. Additionally, we...
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Basement membrane and the role of matrix metalloproteinases during embryonic wound healing
Kadlčíková, Dominika ; Šindelka, Radek (advisor) ; Tolde, Ondřej (referee)
The healing process is an attractive topic in biology and medicine. There are two types of wound healing - in embryos and in adults. In the case of embryonic wound healing the whole process is simplified and accelerated. An inflammatory reaction typical for adult healing is eliminated and the wound is healed without scars. Better understanding of embryonic wound healing could lead to more effective treatment of injuries, burns and chronic wounds in human population. The studying of molecular mechanisms in the healing process is also meaningful in the context of understanding the cancer regulation. The wound healing could be compared with embryonic development in many ways. The body's axes are essential for a spatial activation of genes involved in development. Our hypotesis was that the axis are also important for wound healing. It has been suggested that the cell's polarity and motility depends whether the wound is vertical or horizontal. There are also connections between wound healing and cancer development. One of them is the degradation of the basement membrane (BM) and extracellular matrix (ECM). The process of degradation is catalyzed by the coordinated action of several classes of enzymes. Some of them - matrix metalloproteinases (MMP) and their role in embryonic wound healing have been...
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Healing of wounds during history and nowadays.
ŠVANCÁROVÁ, Kristina
My bachelor thesis "Healing of wounds over course of time" has a theoretical character. Its aim is to analyse the development of wound healing as an integral part of nursing care from prehistoric times up to the present days. This work is a historical-analytical study of primary and secondary literal sources with a following elaboration of an interpretative text. To achieve this it was crucial to set a few research questions: What was the development of wound care from a historical perspective? What influenced wound healing in the historical context? In connection with these research questions it can be stated that the development of wound care started in prehistoric times, where it was influenced by the development of society. Further on it developed in connection with some major events. Among these we can name mostly the influence of Christianity in the Middle Ages, building of medical facilities thanks to the Theresian and King Josef's reforms during the Enlightenment period. Last but not least we have to mention the 19th century which was a breakthrough for the development of wound care due to the new position of women in medicine, nursing as a profession, the establishment of nursing schools and the systematic training of nurses, which continues to develop up to the present days. The historical context that has had an impact on wound healing is characterized in prehistoric and ancient times primarily by the organization of society, religion, and cultural practices that have led to a development of surgery. In the Middle Ages the methods of wound healing stagnated, in comparison with Renaissance, where, thanks to Paré, Mondeville and Paracels, surgery underwent a significant development. The era of Enlightenment and the 19th century were characterized by the development of war surgery, the industrial revolution, the discovery of antisepsis (Lister) and antibiotics (Fleming). Furthermore, the 20th and 21st century were significant due to the discovery of modern materials for wet wound healing and various medical treatments used in wound healing.
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