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Effect of biological treatment and body constitution on in vitro osteoclastogenesis from peripheral precursors in axial spondylarthritisis
Beránková, Martina ; Daňková, Pavlína (advisor) ; Rossmeislová, Lenka (referee)
Introduction: Biological therapy is becoming a common approach to stop disease progression and suppress symptoms of axial spondyloarthritis, an inflammatory rheumatic disease characterized by bone loss due to dysregulation of bone turnover and increased osteoclast differentiation. Obesity likely affects bone metabolism through multiple mechanisms. It can promote adipocyte differentiation and fat accumulation while inhibiting osteoblast differentiation and bone formation. Additionally, obesity is closely associated with chronic inflammation, which can enhance osteoclast activity and bone resorption. This master's thesis aims to examine the potential influence of excessive adiposity on the osteoclastogenesis process among patients receiving biological therapy for axial spondyloarthritis. Materials and methods: Monocytes isolated from the peripheral blood of patients with axial spondyloarthritis undergoing biological therapy and healthy donors were stimulated in vitro with pooled sera from patients receiving biological treatment, patients with different type of treatment, and healthy controls for a duration of 7-14 days. Osteoclasts were evaluated as multinucleated, TRAP-positive cells, and their numbers were subjected to statistical analysis. Markers of bone metabolism and inflammation were assessed...
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Apoptóza mononukleárnych buniek a makrofágov mliečnej žľázy hovädzieho dobytka
Bátik, Andrej
This bachelor thesis is aimed on apoptosis of mononuclear cells and macrophages in bovine mammary gland. The first part is about mononuclear cells. Monocytes can quickly migrate from blood to tissue and differentiate to macrophages. Macrophages are professional phagocytes. They have significant role in immunity respond. These cells react on every antigen and they remove source of this antigen and also remove old and unuseful cells. Next part is about apoptosis. Apoptosis as known as programed cell death. In this part is described morphology, biochemistry and genetics of programed cell death. Apoptosis is completely natural way of dying, without it live cannot exist. Bovine mammary gland is often attacked by staphylococcus and other bacteria. Result of these attacks is in most cases mastitis, inflammatory disease. Mastitis causes serious health danger for cows also causes economical and finance lost in dairy farms. Thesis describes understanding of these processes in order to improve knowledge and battle against inflammatory diseases.
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The significance of extracellular DNA in osteoclastogenesis from peripheral blood precursors - in vitro study
Jelínková, Ivana ; Daňková, Pavlína (advisor) ; Korabečná, Marie (referee)
Introduction: Extracellular DNA (ecDNA) is a common component of blood plasma. Increased levels of ecDNA in plasma can be found in some autoimmune diseases like systemic lupus erythematosus (SLE), rheumatoid arthritis or celiac disease which are associated with inflammatory processes. These diseases are also associated with an increased risk of osteoporosis. Bone is a dynamic structure undergoing constant modelling caused by osteoblasts, osteocytes and osteoclasts. Shifting their equilibrium can lead to pathological conditions such as osteoporosis. In this thesis we focused on elucidating whether ecDNA, an inflammatory agent with proven immunoregulatory effects can alter differentiation potential of monocytes and alternatively lead to osteoclastogenesis via TLR9. Material and methods: We obtained monocytes from peripheral blood of healthy donors and cultivated them with four types of ODNs control (CO), stimulatory (ST), inhibitory (INH, telomeric (TLM) with phosphodiester (-pO) or phosphorothioate (-pS) backbone for two weeks to establish their effect on differentiation potential of monocytes into osteoclasts. Osteoclastogenesis was evaluated by number of yielded osteoclasts observed on a light microscope. To establish the effect of ODNs on osteoclast activity samples were analysed by qPCR for...
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Role of inflammation in pathologic bone resorption in axial spondyloarthritis
Šebová, Eva ; Daňková, Pavlína (advisor) ; Tencerová, Michaela (referee)
Introduction: Axial spondyloarthritis (ax-SpA) is an inflammatory rheumatic disease. It is a unique model of bone remodeling disorders because, although one of the main diagnostic parameters is the rate of bone formation, inflammation present in patients' bodies increases the risk of pathological bone resorption, which can lead to osteoporosis. The processes of pathological resorption in ax-SpA have not been fully investigated, both in the disease as such and in the individual forms of the disease, i.e. non-radiographic (nr-axSpA), radiographic axial spondyloarthritis (r-axSpA) and ankylosing spondylitis (AS). This work deals with the influence of inflammatory serum of patients on the process of osteoclast differentiation from peripheral precursors of patients and healthy donors. Material and methods: Monocytes separated from the peripheral blood of either axSpA patients or healthy donors were stimulated for 14 days in vitro with serum from patients and in parallel with serum of age and sex of the corresponding healthy donors. Osteoclasts were evaluated as multinucleated, TRAP positive cells. Their numbers were statistically processed. Results: The inflammatory serum environment of patients with axSpA stimulated the osteoclastogenesis of axSpA monocytes significantly more (P <0,05) than the...
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Izolace krevních monocytů skotu pro účely kultivace dendritických buněk
Coufalová, Karmela
Monocytes are a population of mononuclear lekocytes. The aim of this thesis was to select a suitable methodology for the isolation of monocytes and their cultivation into dendritic cells. For the experiment, bovine blood was taken from vena jugularis externa. This blood was used to isolate monocytes based on the density gradient of OptiPrep solution and Histopaque solution. The results showed that the method of isolation based on the density gradient of the Histopaque solution and the magnetic separation appeared to be a more efficient method. The monocyte population was cultured for 72 hours. Cells were analyzed by light microscopy and CD14 positive cells appeared to be transformed into dendritic cells. The purity of these cell population was determined by flow cytometry. Dendritic cells have a wide range of utility mainly in immunotherapy of various diseases and also play an important role in cancer therapy.
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The role of monocytes in pathogenesis of diabetes - immunogenetic study
Paukner, Karel ; Daňková, Pavlína (advisor) ; Černá, Marie (referee)
Type one diabetes is an autoimmune disease. It is caused by the destruction of β cells of Langerhans' pancreatic islets. Hyperglycemia is a major symptom of β cell destruction. Monocytes play a key role during T cell activation. T cell effect can be protective (Treg) or destructive. Monocyte destroys β cells as a macrophage and generates self-tolerance as a dendritic cell. The number of patients with T1D is increasing. In the presented work I aim to summarize current information about pathogenesis of T1D and I try to propose future way of research.
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Apoptóza mononukleárnych buniek a makrofágov mliečnej žľázy hovädzieho dobytka
Bátik, Andrej
This bachelor thesis is aimed on apoptosis of mononuclear cells and macrophages in bovine mammary gland. The first part is about mononuclear cells. Monocytes can quickly migrate from blood to tissue and differentiate to macrophages. Macrophages are professional phagocytes. They have significant role in immunity respond. These cells react on every antigen and they remove source of this antigen and also remove old and unuseful cells. Next part is about apoptosis. Apoptosis as known as programed cell death. In this part is described morphology, biochemistry and genetics of programed cell death. Apoptosis is completely natural way of dying, without it live cannot exist. Bovine mammary gland is often attacked by staphylococcus and other bacteria. Result of these attacks is in most cases mastitis, inflammatory disease. Mastitis causes serious health danger for cows also causes economical and finance lost in dairy farms. Thesis describes understanding of these processes in order to improve knowledge and battle against inflammatory diseases.
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