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Anti-inflammatory compounds and stem cells in treatment of spinal cord injury
Kárová, Kristýna ; Jendelová, Pavla (advisor) ; Sameš, Martin (referee) ; Chvátal, Alexandr (referee)
Despite intense scientific efforts, spinal cord injury (SCI) remains to be a severe neurological condition that has no treatment. Currently, therapy is based on alleviating pressure by surgical spinal cord decompression, administration of methylprednisolone and physical therapy. In this study, therapeutic effects of anti-inflammatory compounds and of three types of stem cells were tested in a balloon compression model of spinal cord injury in rats. Natural compounds epigallocatechin-3-gallate (EGCG) or curcumin were administered in situ and then intraperitoneally every day for up to 28 days. Human bone marrow mesenchymal stem cells (MSCs), human spinal neural precursors (SPC-01) and neural precursors derived from human induced pluripotent stem cells (iPS-NPs) were transplanted intrathecally (MSCs) or via spinal injection into immunosuppressed rats 7 days after induction of SCI. To determine effects of therapies, changes in motor function was tested by open field test BBB, flat beam test and score, Plantar test and rotarod. Morphometric analysis was used to assess gray/matter sparing and cavity size. Immunohistochemistry was used to determine survival and differentiation of transplanted cells, activation of classical pathway of NFκB (p65 nuclear translocation), astroglial activation (GFAP) and...
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Surface-modified nanoparticles in the treatment of serious CNS diseases
Mareková, Dana ; Jendelová, Pavla (advisor) ; Králová, Věra (referee) ; Jirák, Daniel (referee)
Glioblastoma multiforme (GBM) is a primary adult brain tumour with an unfavourable prognosis. Standard therapies have a number of side effects, posing a challenge and leading to attempts at new therapeutic approaches. One of these is the use of theranostic nanoparticles with modified surfaces. We have established primary lines from GBM patient samples and characterized them not only in 2D and 3D cultures but also after passaging in the brains of immunodeficient mice. We tested our modified superparamagnetic nanoparticles carrying the drug doxorubicin and RGDS peptides targeting tumor cells on these lines. The γ- Fe2O3@P(HP-MAH)-RGDS-Dox nanoparticles not only reduced cell proliferation in vitro but also incorporated into the tumor and inhibited tumor growth in vivo. We also developed magnetic nanoparticles containing Mn and Zn, which can replace commercially available iron oxide-based contrast agents with their magnetic properties and can be used to label cells for in vivo magnetic resonance imaging. Both in vitro and in vivo tests of the new nanoparticles have revealed that they can be used as a suitable and effective tool for cell tracking using magnetic resonance imaging even where the use of current contrast agents is contraindicated.
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The role of mTOR signalling pathway in neural differentiation of stem cells
Šintáková, Kristýna ; Jendelová, Pavla (advisor) ; Dráber, Peter (referee)
Spinal cord injury is a very serious, complex, and life changing injury for which today's medicine still does not have an efficient treatment. It is only possible to mitigate the consequences of this injury and the pathological processes associated with it. Neural stem cell transplantation has immunosuppressive effects in the pathology of spinal cord injury and promotes regeneration. mTOR kinase is a member of the crucial intracellular PI3K/Akt/mTOR signalling pathway, making it a suitable target for therapeutic intervention and immunosuppressants such as rapamycin. mTOR signalling is important for neural stem cells and in the pathology of spinal cord injury. The aim of this study was to investigate the role of the mTOR pathway in differentiation of stem cells into neuronal phenotype. Rapamycin was applied to in vitro culture of neural progenitors. Immunocytochemistry and immunoblotting techniques were used to study the effect of this inhibition on the cell phenotype and on the activity of the mTOR pathway. Using the rat model of spinal cord injury in vivo, immunohistochemistry and immunoblotting techniques were used to evaluate the impact of rapamycin inhibition on the mTOR pathway, autophagy, and cytokine production by cells in the damaged tissue. The results show that the mTOR pathway plays role...
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Studium interakce buněk s biomimetickým materiálem a jeho využití v biomedicíně
Sauerová, Pavla ; Hubálek Kalbáčová, Marie (advisor) ; Vandrovcová, Marta (referee) ; Jendelová, Pavla (referee)
Biomaterials are considered as very promising tools for regenerative medicine. They have compensatory or supporting function in organism and they are often developed to support specific conventional medical procedures. So-called biomimetic materials are developed to imitate natural environment of organism and to induce positive innate responses of organism. An essential part of biomaterial development is in vitro biological evaluation, which characterizes (often for the first time) the potential of developed material for its clinical application. This Ph.D. thesis deals with in vitro biological evaluation of three different biomimetic materials. In all three cases, the comprehensive evaluation was an integral part of the material development and optimization processes. Each material was in vitro characterised at the level of cell-material interactions with respect to its intended specific application.. In the first part, cell response to potential drug delivery system based on colloidal complexes of cationic surfactants with hyaluronic acid (HyA) was characterized. HyA protection ability and its limits were described; also the role of fetal bovine serum (FBS) in cell response to the stress stimuli was confirmed. Results considered surfactant-HyA complexes as promising system for drug delivery. In...
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Hippocampus Dysfunction in Quinpirole Sensitization Model of Obsessive-Compulsive Disorder
Brožka, Hana ; Stuchlík, Aleš (advisor) ; Jendelová, Pavla (referee) ; Kelemen, Eduard (referee)
Obsessive-compulsive disorder (OCD) is a serious psychiatric condition manifested by repeated thoughts followed by stereotypic compulsive behavior. Alterations to cortico-thalamo-striato- cortical circuits are most often implicated in the pathophysiology of OCD. However, many studies have also found a changed volume, shape and activity of the hippocampus in OCD patients. This work focused on the activity of hippocampal CA1 cells during stereotypical checking behavior and on cognitive flexibility in a quinpirole (QNP) sensitization model of OCD. The activity of CA1 hippocampal cells during stereotypical checking was assessed in an enriched open-field test in QNP sensitized rats. Arc+ (activity-regulated cytoskeletal associated protein, or Arg 3.1) mRNA expression profiles were determined in CA1 coronal hippocampal sections following stereotypical checking. After the establishment of stereotypical checking (10 sessions), rats were exposed to the arena and sacrificed after 5 minutes. QNP sensitized animals visited the same objects with the same frequency as during previous sessions, while control rats did not. Locomotor activity was comparable between QNP treated rats and controls. Following sacrifice, rat brains were flash frozen and sliced to 20 µm thick sections. Sections, mounted on slides, were hybridized...
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The effect of immunosupression on cell therapy in mouse model of Alzeimer's disease
Gajdoš, Roman ; Jendelová, Pavla (advisor) ; Chmelová, Martina (referee)
Alzheimer's disease is a chronic, progressive, neurodegenerative disease. It belongs to the most common type of dementia and worldwide it is statistically the fifth cause of mortality. The most common morphological markers are insoluble β amyloid plaques, hyperphosforylated tau proteins and formation of neurofibrilar tangles. Among the manifestations of the disease is amyloid angiopathy, synaptic transmission disorders and subsequent apoptosis, deterioration of cognitive functions and brain atrophy. Studies have shown that administration of mesenchymal stem cells (MSC) has an immunomodulatory effects and it can reduce the production and storage of β amyloid and thus improve cognitive functions. In preclinical studies, which are conducted in transgenic mice and often use xenografts, administration of immunosuppresion may lead to variety of positive or negative effects which can affect the results of the experiment. The subject of the master's thesis was to determine the effect of immunosuppression on experimental therapy with MSC in various time windows of AD progression (model 3xTg). At which scale and combination of immunosupression will influence the cell therapy's effects, the length of graft survival, mortality of experimental animals and changes at the cellular level. We have also assessed...
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The use of stem cells in the treatment of Amyotrophic lateral sclerosis
Řehořová, Monika ; Jendelová, Pavla (advisor) ; Klíma, Jiří (referee) ; Jiruška, Přemysl (referee)
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease caused by degradation of motoneurons (MN). No effective treatment is currently available. Cellular therapy is considered to be a promising experimental treatment that could target the pathology of complex disease through many potential mechanisms. We compared the effect of three types of repeated applications of human mesenchymal stem cells (hMSC): intrathecal administrations, intramuscular administrations and the combination of these applications. Best results were obtained after combined repeated hMSC administrations. We observed the rescue of MN, neuromuscular junctions and decreasing levels of proteins involved in the signaling of necroptosis (Rip1, cl-casp 8), apoptosis (cl-casp 9) and autophagy (beclin 1), decreasing astrogliosis and the level of astrocytic connexin 43. Neural precursors derived from induced pluripotent stem cells (NP-iPS) are considered as other promising candidates for ALS therapy. Intraspinal administration of NP-iPS increased mRNA expression of BDNF and IGF-1, on the other hand decreased expression of proapoptotic casp 3. We also observed their effect on expression of components of perineural nets (PNN). Human embryonic stem cells (hESC) are the other cell candidates for the treatment of neurodegenerative...
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Treatment of spinal cord injury targetting secondary mechanisms
Valášková, Barbora ; Jendelová, Pavla (advisor) ; Hájek, Milan (referee) ; Rokyta, Richard (referee)
1 Abstract Traumatic spinal cord injury (SCI) is severe condition with devastating long- lasting consequences. There is still no available treatment. After initialing mechanical trauma, there is a huge cascade of secondary reactions which amplifies the damage. This thesis is focused on potential therapeutic effect of photobiomodulation, natural anti- inflammatory compounds epigallocatechin-3-gallate (EGCG), curcumin, their combination and extrapure synthetic form of curcumin called "nanocurcumin" on impacts of experimental model of SCI in rats. Photobiomodulation using combination of two synchronized wavelengths 808 and 905 nm improved functional recovery after SCI evaluated by battery of locomotor tests and somatosensory plantar test. The group treated by photobiomodulation obtain better results in all tests. The histopathological analysis showed a positive effect on white and gray matter sparing and our data suggests an upregulation of M2 microglia/macrophages in photobiomodulation treated rats assessed by immunohistochemical and RT-qPCR analysis. Our results demonstrated that the photobiomodulation is a promising non-invasive therapy for improving functional recovery and tissue sparing after SCI. EGCG and curcumin are natural compounds known in Chinese medicine for centuries. Their neuroprotective and...
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Modulation of memory using enzymatic digestion of perineuronal nets in perirhinal cortex
Procházková, Natálie ; Jendelová, Pavla (advisor) ; Chmelová, Martina (referee)
In the adult brain, neuronal plasticity is regulated by a specialized structure of extracellular matrix, the perineuronal nets (PNNs), which restrict synaptic plasticity by binding molecules of inhibitory nature and posing as a physical barrier to alterations in neuronal connectivity. This effect is abolished by removal of PNNs by the enzyme chondroitinase ABC (chABC), which enables to reopen critical period window and leads to memory improvement. Here, we utilized chABC and a novel approach in removing the PNNs in perirhinal cortex, using the enzyme hyaluronidase (Hyase), to assess differences in the use of these enzymes in object recognition (OR) memory improvement and alterations in the structure of neuronal network of wild type mice. Our findings suggest that Hyase may be a more convenient tool to PNN removal than chABC, as Hyase surpasses chABC in promoting the OR memory, influence larger portion of neuronal network by affecting both inhibitory and excitatory neurons, and provides extended temporal window for experimental modulation of activity-dependent synaptic plasticity. Key words: perineuronal nets, synaptic plasticity, chondroitinase ABC, hyaluronidase, perirhinal cortex, memory
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