Mesenchymal stem cells (MSCs) are a type of stem cells that are important for the development and regeneration of the body's musculoskeletal system. They act as precursor cells and can differentiate into adipocytes, osteoblasts, and chondrocytes. Adipogenesis and osteogenesis are constantly taking place in the body, many processes and signaling pathways that stimulate one and inhibit the other and vice versa. In old age, an imbalance between the two processes occurs, namely the rate of differentiation of MSCs into adipocytes instead of osteoblasts increases. This can lead to diseases such as osteoporosis, which is characterized by increased bone fragility due to reduced bone mass and changes in bone microarchitecture.
Greater bone building is stimulated by the osteoanabolic agent teriparatide, which we studied in our research work. The aim of the master's thesis was to determine the differences in the expression of PPARG and FABP4 genes, which participate in the cascade of MSC adipogenic differentiation processes. We used 41 MSC samples that underwent adipogenic differentiation and were continuously or intermittently exposed to teriparatide. It mimics the action of parathormone, which stimulates bone formation with intermittent application and bone breakdown with continuous application. Using different methods of teriparatide application, we wanted to determine their influence on the adipogenic differentiation of MSCs.
First, RNA was isolated from MSC samples and transcripted into cDNA. Then, we measured the expression of adipogenic markers using the quantitative real-time polymerization chain reaction method. The success of adipogenic differentiation was also determined by histochemical staining of adipocytes. For both target genes, we detected the highest expression in samples that were continuously exposed to teriparatide for 7 days, which was also in accordance with the results of adipocyte staining. They showed that adipogenesis was successful only in samples that were continuously exposed to teriparatide and in samples that were not exposed to teriparatide. We did not measure any major differences in the expression of the PPARG gene between intermittent and continuous administration of teriparatide. In the expression of the FABP4 gene, the same pattern of expression was observed for all methods of teriparatide application.
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