The Regulation Of Intervertebral Disc Cell Interactions With Their Surrounding Microenvironment
Hsieh, Adam H
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Intervertebral disc degeneration is the major cause of back pain in the US, which can be both physically debilitating and costly to treat. Current treatments include invasive surgeries, which can be effective in ameliorating pain, but also contain the risk of complications. Additionally, these strategies target clinical manifestations of disc degeneration, rather than examine the cause of degenerative changes. Therefore, current research focuses on finding minimally invasive treatments for disc disease such as gene therapy. Regulating intervertebral disc cell interactions with their immediate environment can be a useful tool in the development of therapeutic strategies. This was explored through environmental changes to assess shifts in cell phenotype as well as genetic modulation to elucidate alterations in cell function. Biochemical, nutritional, and physical factors were examined in immature nucleus pulposus cells to assess changes in gene expression, attachment, and proliferation. It was found that nutritional and physical factors can alter gene expression levels of NP cells, thereby altering cell phenotype. In addition, down-regulation of the proteolytic enzyme MMP-2 was explored through RNAi interference. Five shRNA lentiviral vectors were designed and validated for the sustained gene silencing of MMP-2. Silencing MMP-2 activity resulted in the inability of disc cells to focally degrade gelatin films as well as reduced ability of disc cells to remodel fibers in type I collagen gels, resulting in weakened gel architecture. These functional consequences were further explored in an in vivo study utilizing an annular needle-puncture model of disc degeneration. Injection of the shMMP lentiviral construct lead to decreased expression of MMP-2 in the disc, as well as improved disc height and morphology. Thus, the functional consequences of silencing MMP-2 were examined, elucidating its role in the degradative pathway leading to degenerative disc disease. The results of these studies can lay the foundation for developing therapeutic treatments for intervertebral disc degeneration.