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Mesenchymal stromal cells and biological scaffolds for neural tissue regeneration
Kočí, Zuzana ; Kubinová, Šárka (advisor) ; Filová, Eva (referee) ; Zach, Petr (referee)
Despite tremendous progress in medicine, injuries of the adult central neural system remain without satisfactory solution. Regenerative medicine employs tissue engineering, cellular therapies, medical devices, gene therapy, or growth factors with the aim to bridge the lesion, re-establish lost connections and enhance endogenous repair in order to restore neural function. The aim of my thesis was to evaluate therapeutic potential of two approaches, transplantation of human mesenchymal stromal cells (hMSCs) and biological scaffolds derived from extracellular matrix (ECM) for neural regeneration, particularly in models of spinal cord injury (SCI). First, hMSCs from various sources - bone marrow (BM), adipose tissue (AT) and Wharton's jelly (WJ) - were isolated and characterized in vitro. All cell types met the minimal criteria for MSC phenotype and displayed similar properties in terms of their surface marker expression, differentiation potential, migratory capacity, and secretion of cytokines and growth factors. On the other hand, the cell yield from WJ and AT was significantly higher, and MSCs isolated from these tissues proliferated better than from BM. Therapeutic effect of intrathecal application of hWJ-MSCs was then evaluated in SCI compression model in rats. The effect of low (0.5 million) and...
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Development of extracellular-matrix scaffolds for CNS repair
Výborný, Karel ; Kubinová, Šárka (advisor) ; Hampl, Aleš (referee) ; Vandrovcová, Marta (referee)
Výborný, Karel. Vývoj materiálů na bázi extracelulární matrix pro léčbu centrálního nervového systému. [Development of extracellular-matrix scaffolds for CNS repair]. Praha, 2020. 105 stran, 3 přílohy. Dizertační práce (Ph.D.). Univerzita Karlova, 2. lékařská fakulta, Ústav experimentální medicíny, AV ČR v.v.i. Abstract Brain and spinal cord injury are a serious traumata and despite intensive research, there is still no effective treatment for patients. One of the studied approaches is to use various biomaterials to repair the damaged neural tissue. The aim of this thesis is to study the regenerative and neurotrophic effects of injectable extracellular matrix (ECM) hydrogels prepared by decellularization from porcine tissue (brain, spinal cord, bladder) and human umbilical cord (UC) in reconstruction of damaged neural tissue of the brain and spinal cord in rats. We characterized ECM hydrogels in terms of mechanical and rheological properties, structure and composition, cell adhesion, migration and proliferation. In a model of spinal cord hemisection, we compared the regenerative effect of ECM hydrogels derived from porcine spinal cord and urinary bladder. We found no benefits of tissue specific ECM prepared from the tissue of neural origin in terms of its neurotrophic properties in vitro or in vivo....
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Natural biomaterials and mesenchymal stem cells in regeneration of spinal cord injury
Kekulová, Kristýna ; Kubinová, Šárka (advisor) ; Krulová, Magdaléna (referee) ; Filová, Elena (referee)
Spinal cord injury is a serious trauma and despite intensive research there is still no effective treatment for patients. The aim of this thesis is to study new possibilities of spinal cord injury therapy in animal models. We have focused on the use of natural materials, stem cells, gene therapy and the possibility of combining these approaches. The effect of extracellular matrix (ECM) based materials prepared by decellularization of porcine spinal cord and porcine urinary bladder on tissue regeneration after acute hemisection of the spinal cord was investigated. Another tested material was a hydrogel based on hyaluronic acid modified with RGD adhesion peptide, which was applied acutely and subacutely into the hemisection lesion. We have shown that both types of biomaterials have positive effect on regeneration of the spinal cord tissue by bridging the lesion and promotion of axonal ingrowth. In addition, ECM hydrogels promote the growth of blood vessels into the lesion site. The combination of hydrogels with mesenchymal stem cells derived from human umbilical cord (hWJ-MSCs) had synergistic effect, but since only a limited number of cells could be incorporated into hydrogels, this effect was not associated with improvement in motor skills. The limitation of ECM hydrogels is their rapid...
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Effect of endogenous factors on mesenchymal stem cells
Černá, Kristýna ; Krulová, Magdaléna (advisor) ; Kubinová, Šárka (referee)
Maintaining of homeostasis is essential for the survival of the organism. Stress disturbs the homeostasis and prepares the organism for mental or physical stress. During the stress situation, the endogenous stress factors are released. Through these factors stress affects tissue regeneration, the immune system and other metabolic processes. Chronic stress impacts many parts of body and mind and has a negative effect on these processes. Acute stress has the opposite effect. Mesenchymal stem cells (MSCs) participate in regenerative processes and modulate the immune system. Therefore, it can be assumed that stress will affect on MSCs. The aim of this study was to investigate the effect of stress factors, norepinephrine and corticosterone on the properties and function of MSCs in acute and chronic stress model. In our study, stress factors did not affect the morphology, vitality and differentiation of MSCs. However, the metabolic activity of MSCs was reduced regardless of the duration of their action. The action of stress factors also affected the production of some immunologically relevant molecules and proteins. Unfortunately, the results did not show a clear effect of stress factors on the lymphocyte modulation by MSCs. Key words: mesenchymal stem cells, catecholamines, adrenergic receptors,...
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Mineral and chemical changes of magma crystallization during formation of post-Variscan intrusions and veins in the Moldanubian Zone of the Bohemian Massif
Kubínová, Šárka ; Faryad, Shah Wali (advisor) ; Rapprich, Vladislav (referee) ; Moyen, Jean-François (referee)
- 1 - English abstract The late-Variscan magnesium-rich potassic to ultrapotassic igneous rocks create numerous dykes, dyke swarms and several plutonic bodies at the boundary between the Moldanubian Zone and the Teplá-Barrandian Block of the Bohemian Massif. They represent a volumetrically smaller group of igneous rock but they are the key to understand generation of melt and processes of mantle metasomatism and shallow-level magma differentiation. In addition, they are considered as an indicator of the tectonic evolution of this part of the Bohemian Massif during the final stages of the Variscan orogenesis. Currently, they are the subject of discussion by several authors in terms of their genesis, emplacement time and geodynamic significance. The presented thesis is a compilation of four scientific publications that are aimed at the petrological study of selected (ultra)-potassic dyke rocks from several localities at the western border of the Moldanubian Zone. The study of mineral textures, mineral chemistry and whole-rock geochemistry together with magnetic fabrics, structural field relations and age determinations allowed us to describe the crystallization history of these rocks, discuss their evolution from melt generation to magma ascent and emplacement, and form the model of sequence of Variscan...
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Mesenchymal stromal cells and biological scaffolds for neural tissue regeneration
Kočí, Zuzana ; Kubinová, Šárka (advisor) ; Filová, Eva (referee) ; Zach, Petr (referee)
Despite tremendous progress in medicine, injuries of the adult central neural system remain without satisfactory solution. Regenerative medicine employs tissue engineering, cellular therapies, medical devices, gene therapy, or growth factors with the aim to bridge the lesion, re-establish lost connections and enhance endogenous repair in order to restore neural function. The aim of my thesis was to evaluate therapeutic potential of two approaches, transplantation of human mesenchymal stromal cells (hMSCs) and biological scaffolds derived from extracellular matrix (ECM) for neural regeneration, particularly in models of spinal cord injury (SCI). First, hMSCs from various sources - bone marrow (BM), adipose tissue (AT) and Wharton's jelly (WJ) - were isolated and characterized in vitro. All cell types met the minimal criteria for MSC phenotype and displayed similar properties in terms of their surface marker expression, differentiation potential, migratory capacity, and secretion of cytokines and growth factors. On the other hand, the cell yield from WJ and AT was significantly higher, and MSCs isolated from these tissues proliferated better than from BM. Therapeutic effect of intrathecal application of hWJ-MSCs was then evaluated in SCI compression model in rats. The effect of low (0.5 million) and...
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Clinical applications of stem cells for the treatment of neurodegenerative diseases of CNS
Jančová, Pavlína ; Kubinová, Šárka (advisor) ; Heřmánková, Barbora (referee)
Stem cells have a huge therapeutic potential due to their ability to differentiate in multiple tissues. They could be used for neurodegenerative diseases treatment, which are typical for loss of specific groups of neurons, progressive course and lack of effective treatment due to their complicated pathophysiology, only therapies for elongation and simplification of patients' life are available. This thesis summarizes results of completed clinical studies and informs about ongoing studies, in which stem cell treatments are used for selected neurodegenerative diseases. Stem cell therapy for multiple sclerosis and amyotrophic lateral sclerosis have a long history, some of the studies has proven therapeutic efficiency of stem cells. We don't know much about effect of stem cell treatment for patients with Alzheimer's and Parkinson's diseases, because first clinical studies were finished recently. But all clinical trials have proven safety of stem cell treatment. Replacement of damaged neurons haven't been reached yet, just protection of remaining neurons by neurotrophic and immunomodulatory factors secreted by stem cells. Keywords: neurodegenerative diseases, stem cells, clinical trials, multiple sclerosis, amyotrophic lateral sclerosis, Alzheimer's disease, Parkinson's disease
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Methodology of using low-temperature plasma in veterinary treatment
Dejneka, Alexandr ; Kubinová, Šárka ; Moravec, J. ; Bezděk, D.
The aim of the methodology is application of low-temperature plasma in veterinary treatment. Low-temperature plasma (NTP) has non-specific antimicrobial effects and it can be safely applied to living tissues without damaging them. The methodology presents the solution of dermatological problems with potential bacterial, fungal or yeast contamination. The method can also be used for antimicrobial decontamination and for the promotion of healing of chronic and non-healing wounds.
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Analýza volných prostorů brněnských panelových sídlišť
Kubínová, Šárka
The article discusses itemized dissertation on “Open space of prefab housing estates in Brno”. The aim of the thesis is to analyse and describe the unused premises of panel house estates in Brno and to create and valorise usable system of data and information (regarding chronological sequence). The obtained data and the entire dissertation can be used in planning and designing interventions in prefab housing estates and in other structures of residential areas, respectively, as part of a multi-criteria analysis of particular housing estates.
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Immunogenicity of stem cells and their derivatives
Doležalová, Nikola ; Holáň, Vladimír (advisor) ; Kubinová, Šárka (referee)
Immunogenicity of stem cells and their derivatives Nikola Doležalová Abstract Stem cells (SCs) have the potential to be used in regenerative medicine on the basis of their differentiation capacity and promising immunological properties, including low expression of histocompatibility antigens and costimulatory molecules, or secretion of suppressive cytokines. Their immunogenicity has often been ignored in the past but it is becoming clear that rejection of genetically incompatible SCs represents a very common issue. At present, SCs are extensively studied from the immunological point of view, since it represents a critical aspect of the safety of SC therapy. This thesis presents an overview of current knowledge about immunogenicity of SCs and their derivatives, including both pluripotent SCs (embryonic and induced pluripotent SCs) and adult SCs (mesenchymal, limbal, neural, haematopoietic and umbilical cord blood SCs). The expression of immunologically relevant molecules on their surface and interaction with the immune cells in vitro and in vivo will be discussed, together with suggestions for overcoming the immunological barriers for transplantation. Detailed analysis of these aspects necessarily has to precede the safe clinical translation of SC therapies.
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