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Development of Material Based on Hyaluronic Acid s Hydrogels for Myocardial Regeneration
Kovářová, Lenka ; Kubala,, Lukáš (referee) ; Prokš,, Vladimír (referee) ; Pekař, Miloslav (advisor)
The thesis is focused on material development based on hyaluronic acid usable in regenerative medicine, especially for heart tissue regeneration after myocardial infarction. The object of the study is the oxidized form of hyaluronic acid (HA-Ox) and hydroxyphenyl derivative of HA (HA-TA). HA-Ox can be crosslinked with a bifunctional alkoxyamine POA and HA-TA undergoes an enzymatic reaction in the presence of hydrogen peroxide catalysed by horseradish peroxidase leading to gel formation. To describe the materials, chemical and physical properties, gelation kinetics and conditions of crosslinking reactions were studied. Hydrogels were characterized by mechanical and viscoelastic properties, degradability or stability in simulated body fluids. These hydrogels serve as scaffolds for the selected cell type. To promote cell adhesion and viability, an RGD sequence has been bonded to the structure of HA-TA. This resulting material is also compatible with selected applicators. Its viscosity and extrusion force are low enough to allow application with a catheter with a very small internal diameter. The applicability of the material through the supply tube to the hydrogel reservoir of the second SPREADS device showed good homogeneity, cell distribution and viability. Finally, the material was applied in vivo using these devices during a preclinical study.
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Immunomodulatory potential of Sertoli cell progenitors during heart regeneration after injury in Xenopus
Žabková, Světlana ; Krylov, Vladimír (advisor) ; Procházka, Jan (referee)
The high percentage of mortality associated with cardiovascular diseases shows the importance of studying cardiac muscle regeneration. An interesting approach for heart regeneration may be the use of a cell culture of Sertoli progenitor cells and peritubular myoid cells derived from juvenile testes of Xenopus tropicalis that was named XtiSC. These cells were found to share a number of characteristics which are typical for mesenchymal stem cells. Mesenchymal stems cells are well-known for their immunomodulatory and anti-inflammatory abilities that promote heart regeneration. The goal of this master thesis was to test the immunomodulatory potential of Sertoli progenitor cells during heart regeneration after heart apex amputation in Xenopus tropicalis without macrophages. The reason for macrophages depletion was the assumption that XtiSC behave like macrophages in many aspects. Clodronate encapsulated in liposomes was chosen as a tool for macrophage depletion. It was confirmed that XtiSC can positively influence heart regeneration in a classic model of heart regeneration, but also in a model with depletion of macrophages. It was revealed that Xenopus tropicalis adults without macrophages injected with XtiCS into the hindlimb skeletal muscle 3 days before apical resection showed reduced fibronectin...
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Development of Material Based on Hyaluronic Acid s Hydrogels for Myocardial Regeneration
Kovářová, Lenka ; Kubala,, Lukáš (referee) ; Prokš,, Vladimír (referee) ; Pekař, Miloslav (advisor)
The thesis is focused on material development based on hyaluronic acid usable in regenerative medicine, especially for heart tissue regeneration after myocardial infarction. The object of the study is the oxidized form of hyaluronic acid (HA-Ox) and hydroxyphenyl derivative of HA (HA-TA). HA-Ox can be crosslinked with a bifunctional alkoxyamine POA and HA-TA undergoes an enzymatic reaction in the presence of hydrogen peroxide catalysed by horseradish peroxidase leading to gel formation. To describe the materials, chemical and physical properties, gelation kinetics and conditions of crosslinking reactions were studied. Hydrogels were characterized by mechanical and viscoelastic properties, degradability or stability in simulated body fluids. These hydrogels serve as scaffolds for the selected cell type. To promote cell adhesion and viability, an RGD sequence has been bonded to the structure of HA-TA. This resulting material is also compatible with selected applicators. Its viscosity and extrusion force are low enough to allow application with a catheter with a very small internal diameter. The applicability of the material through the supply tube to the hydrogel reservoir of the second SPREADS device showed good homogeneity, cell distribution and viability. Finally, the material was applied in vivo using these devices during a preclinical study.
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