Mesenchymal stem/stromal cells are a heterogeneous population of stem cells that are found in adult tissues and have the unique ability of multilineage differentiation, in particular the differentiation within the mesoderm. During embryonic development connective tissues such as cartilage, muscles, fat and others develop from the mesoderm. Mesenchymal stem/stromal cells (MSCs) have been the most common research target in the field of regenerative medicine for some time. Their physiological and pathophysiological role in degenerative disorders of the hip shows their potential use in the diagnosis of these disorders. The most common degenerative disorders of the musculoskeletal system are osteoporosis, osteoarthritis (OA) and sarcopenia. OA is a diverse and multifactorial disorder of the synovial joints, where inflammatory factors are also involved. The basis of the disease is the progressive loss of the structure and repair function of the joint through the endogenous regenerative process, which is suggested to be weakened in the aforementioned disease. Namely, in younger healthy individuals, the regenerative process after tissue damage is carried out by MSCs. The aim of the master's thesis was to determine differences in the expression of genes encoding mediators of inflammation in MSCs obtained from different hip tissues of subjects with and without degenerative changes. During arthroscopy of the hip, synovium at 3 different locations of the hip (S1-S3) and a pillar of the trabecular bone of the femur (KO) were taken from the subjects. Primary MSCs were then isolated from these tissues and cultured in vitro and RNA was isolated from culture expanded cells. We then optimized a quantitative polymerase chain reaction (qPCR) method to measure the expression of genes for inflammatory mediators. Based on the optimization results, we selected TGFB1, IL6, COMP, MMP13 and VEGFA genes for further analysis. The results obtained by qPCR were normalized to reference gene GAPDH. The results were then compared between patients with early OA and those without, and between different tissues. We tested 3 hypotheses, namely: gene expression of inflammatory molecules differs between MSCs from patients with early OA compared to those without OA; gene expression of inflammatory molecules differs between MSCs from different tissues of patients with early OA and gene expression of inflammatory molecules differs between MSCs from different tissues of patients without OA. Statistical analysis of the results did not show statistically significant differences in the expression of the investigated genes, so we rejected all 3 hypotheses. Based on these results we can conclude that MSCs from different tissues of the hip and patients with OA and those without hip degenerative changes do not differ in the expression of genes for inflammatory molecules. The differences between these MSCs are probably result of other molecules. The limitation of this study is also small group of samples, so we suggest further research in larger population in order to increase the power of the statistical test.
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