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Smart biomaterials for connective tissues regeneration
Hefka Blahnová, Veronika ; Filová, Eva (advisor) ; Maxová, Hana (referee) ; Motlík, Jan (referee)
Connective tissues are characterized by significant volume of extracellular matrix. Their main role is to provide a mechanical support and protection to other body organs. This thesis is focused on regeneration of bone, cartilage and osteochondral defect. In the experimental part we observed viability and differentiation of human mesenchymal stem cells. In vitro we evaluated the potential of PCL scaffold with addition of growth factors, bone xenograft with biomimetic peptides, collagen I composite with bioceramics and a titanium alloy with nanostructured surface. During following in vivo study, we implanted a cell-free scaffold made of PCL, calcium phosphate and IGF-1, bFGF, TGFβ1 and BMP-2 to osteochondral defect. Unfortunately, addition of growth factors resulted in pathological inflammatory process despite clear beneficial effect in vitro. Likewise, the biomimetic peptide sequences promoted osteogenic differentiation of human mesenchymal stem cells. Addition of certain bioceramics influenced the scaffold morphology in the manner of pore size. However, we did not observe any effect of the surface characteristics on cell behavior. The cells were influenced rather by certain material. On the other hand, surface modification of titanium scaffold by anodic oxidation revealed that the most suitable...
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Genetic regulation of the cranial cartilage and bone development
Burianová, Andrea ; Machoň, Ondřej (advisor) ; Kaucká, Markéta (referee)
The skull development is an elaborate sequence of cellular processes, featuring two distinct tissue lineages, a unique transient pluripotent population of neural crest cells and mesodermal cells. Several differences in molecular mechanisms operating during chondrogenesis and skeletogenesis have been demonstrated in cranial bones originating from these different cell lineages. The bones comprising the cranial skeleton are formed through both intramembranous and endochondral ossification, regardless of the origin. The cellular processes involved in the formation of cranial skeletal elements include induction, migration, condensation, differentiation, and proliferation, all of which require sophisticated genetic control. Recent discoveries provide evidence of several signaling pathways and their target genes contributing to cranial skeleton development. The interconnection between individual signaling cascades is extremely complex and creates an entire gene regulatory network. This thesis focuses primarily on genetic programmes controlling the development of neural crest-derived skeletal structures. Keywords: neural crest cells, osteogenesis, chondrogenesis, mesenchymal condensation, gene regulatory network
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Coculture of cells in vitro for bone regeneration
Sloupová, Lenka ; Filová, Eva (advisor) ; Tlapáková, Tereza (referee)
Cocultivation of two (or more) cell types in vitro leading to the formation of functioning bone tissue, later inserted into the damaged area, could be a solution for patients for whom the current methods (e.g. use of human bone grafts) are insufficient. In coculture, one cell type is used to accomplish osteogenesis, while the other is used for angiogenesis, because the limit of diffusion of O2 and essential nutrients is only 200 μm, which means that establishing a vascular network in vitro should prevent the new bone tissue from dying after implantation. Creation and understanding of a functioning coculture in vitro are crucial for developing a coculture successful in vivo. This work summarises and compares information about the influence of in vitro cocultivation on proliferation, osteogenesis and angiogenesis in coculture which uses osteoblasts (or osteoprogenitors), bone marrow mesenchymal stem cells (BMSC) or adipose derived mesenchymal stem cells (ADSC) as it's osteogenic cell type combined with various endothelial cell types. In order to understand the impact of cocultivation on these processes, one chapter deals with interactions between cocultured cell types. Keywords coculture, osteogenesis, angiogenesis, in vitro, osteoblasts, BMSC, ADSC
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Osteogenesis and bone healing in congenital short femur
Frydrychová, Monika ; Dungl, Pavel (advisor) ; Havlas, Vojtěch (referee) ; Charvát, Jan (referee)
Introduction: Congenital short femur, or proximal femoral focal deficiency (PFFD), is a rare complex deformity of the lower extremity with femoral dominance. The clinical findings cover wide range of variety, from femoral absence till inconspicuous shortening of the femur. Aim of the study: 1. Molecular analysis of pseudoarthrosis tissue in congenital short femur with focusing on osteogenic and angiogenic gene expression in comparison with physiological bone. The differences in gene expression were expected. 2. Retrospective analysis of femoral healing after prolongation calculating the severity of affection, age, distance of elongation and complication. The extended healing according to severity type and age was expected compared to control group. Material and methods: The RNA from piece of one was isolated and transcription profile of possible 113 genes of osteogenesis and angiogenesis was detected by biochip technology (SuperArray Bioscience Corporation). 10 samples analyses were performed (7 of PFFD, 3 controls). The data of 57 PFFD patients indicated for elongation of the femur with the types Pappas III, IV, VII, VIII and IX and 12 patients in control group were evaluated retrospectively and statistically by GLS method. Results: The expected differences in gene expression in PFFD tissue...
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