| Abstract [eng] |
As life expectancy rises, so does the incidence of age-related diseases. Cartilage tissue of joints has a limited capacity to regenerate after physical trauma, ageing processes and degenerative diseases such as osteoarthritis (OA). Currently, there is no widely used and available drug that stops the degen-eration of cartilage and heals its damaged structure. Different methods, such as stem cell therapy, are being explored to create healthy cartilage tissue from differentiated cells that can be transplanted onto lesions. Cartilage created by current techniques often does not have the same characteristics as carti-lage found naturally in the body, so it is important to develop technologies that improve the engineer-ing of cartilage tissue for therapeutic purposes. Cell sheet technology is a technique based on culturing cells on a thermosensitive polymer, PIPAAm, which allows temperature changes to be used to detach cell sheets as structures containing all the components necessary for tissue biointegration after trans-plantation. Chondrogenically differentiated stem cell sheets can be directly applied to surface articular cartilage lesions or degeneration. Physiological hypoxia is the concentration of oxygen naturally present in the body, which varies between different tissues and organs. One of the main ways in which cells continue to grow and adapt under physioxic conditions is through hypoxia-inducible factors (HIFs) – transcription factors that are activated in the presence of a limited amount of oxygen. They induce changes in gene expression related to cellular adaptation, metabolism and maintenance of homeostasis. In cartilage, this tissue is crucial for chondrogenesis, and studying their mechanisms can be applied to the development of differentiated cartilage tissue of MSC origin. In this paper, the effect of 5 % O2 physioxia conditions on bone marrow mesenchymal stem cells (BMSCs) and articular chondrocytes was investigated in comparison to laboratory normoxia (20 % O2) with the addition of the HIF-1α inhibitor LW6. In this paper, successful chondrogenically differentiated cell sheets were grown, we demonstrat-ed the benefits of physioxia on cell proliferation and chondrogenesis-related gene expression, de-scribed changes in aerobic and anaerobic metabolism, intracellular calcium ions, cell migration and investigated the mechanisms and targets of inhibition of LW6 itself by examining the intracellular proteins HIF-1α, VHL, and the activity of the enzyme MDH2 in conditions of physioxia with LW6. |
