THE EFFECTS OF 3,4,6-O-TRIBUTANOYLATED-N-ACETYLGLUCOSAMINE ON INFLAMMATION AND METABOLISM IN MESENCHYMAL STEM CELLS AND CHONDROCYTES

osteoarthritis;metabolism;inflammation;sugar analog;chondrocytes;mesenchymal stem cells;Biomedical Engineering

Ramesh, SandhyaElisseeff, Jennifer H. ;

Johns Hopkins University

关键词: osteoarthritis;    metabolism;    inflammation;    sugar analog;    chondrocytes;    mesenchymal stem cells;    Biomedical Engineering;   

Others  :  https://jscholarship.library.jhu.edu/bitstream/handle/1774.2/38067/RAMESH-THESIS-2014.pdf?sequence=1&isAllowed=y

瑞士|英语

来源: JOHNS HOPKINS DSpace Repository

【 摘 要 】

Osteoarthritis, or degenerative joint disease, affects over 27 million adults in the United States. It is characterized by a net loss of cartilage tissue in the joints, resulting in pain, swelling, and loss of mobility. Chondrocytes are the resident cell type of articular cartilage, and due to their low mitotic activity and the lack of blood vessels or nerves in the tissue in which they reside, cartilage has a limited ability to self-repair. There are currently no approved treatments to successfully halt or reverse the progression of the disease. In order to address the resulting need for an effective therapeutic, previous work has discovered a novel sugar analog, 3,4,6-O-Bu3GlcNAc, which has shown the ability to reduce inflammation and increase tissue production in an in vitro model of inflammation in animal cells. The following studies seek to expand on those results by first showing that the analog is also effective on human cells. When the analog was cultured with both human mesenchymal stem cells and human OA chondrocytes stimulated by IL-1β to mimic the inflammatory conditions of osteoarthritis, expression of genes related to the inflammatory pathway decreased as evaluated by real-time PCR. In addition, genes related to the production of the extracellular matrix increased. Thus, we were able to show that the analog produced comparable effects in human cells. Subsequent studies focused on characterizing the bioenergetics profile of mesenchymal stem cells during differentiation, inflammation, and treatment with the sugar analog. It was found that for most conditions, after an initial increase, the differentiating cells displayed lower levels of oxygen consumption rates than proliferating cells. In monolayer conditions, human and goat MSCs had differing ratios of oxidative phosphorylation to glycolysis in the absence of the analog, while the addition of the analog eliminated this difference. In three dimensional conditions, except for the unstimulated human MSCs, neither inflammation nor the analog produced any significant differences in oxygen consumption rates. However, the unstimulated human MSCs did not display the same trend and showed an initial difference between the analog-treated and untreated conditions that slowly decreased over the course of differentiation. These results present information that may be useful when trying to determine the mechanism of effect of the sugar analog or develop other therapeutics for the treatment of the disease. In conclusion, we have shown that the sugar analog has comparable effects in human cells that demonstrate that it has the potential to be an effective drug candidate for the treatment of osteoarthritis, and furthermore, we have characterized some of the metabolic effects of the drug to begin to understand the mechanisms behind its effects or possibility to aid in future design.