Breakdown of the disc extracellular matrix is thought to arise from increased activity of matrix metalloproteinases (MMPs). Aggrecan, one of the major disc matrix macromolecules, is degraded through action of MMPs and aggrecanases and its concentration falls early in the degeneration process. Loss of the constituent glycosaminoglycans (GAGs), which are osmotically active, leads to a decrease in osmotic pressure and subsequently to a fall in tissue hydration. Apart from the major biomechanical consequences, fall in extracellular osmolarity is known to influence cell function. Apart from aggrecan, other macromolecules such as fibronectin are also broken down by MMPs. Fibronectin fragments (Fnfs) have been identified in degenerate discs. Such Fnfs have been found to stimulate production and activity of MMPs in articular cartilage but little is known about the effect of these fragments on disc cells. The aim of the work was thus to determine whether Fnf stimulated expression of MMPs and hence induced aggrecan breakdown and loss in the disc and whether extracellular osmolarity influenced this potential response. NP cells or explants were harvested from adult bovine caudal discs. They were cultured in DMEM culture medium over a range of osmolarities with or without Fnf treatment. Profiles of gene expression of MMPs and their inhibitors and effect of changes in osmolarity on expression of selected MMPs were determined. The effect of Fnf on responses of cells and tissue explants from the central region of the disc, the nucleus pulposus (NP) and the role of changes in extracellular osmolarity in relation to GAG loss and expression of selected MMPs was then examined both at the protein level and by gene profiling using a microarray. My results showed that expression of MMPs by disc cells is regulated by extracellular osmolarity rather than the 30 kD Fnf, with the level of some MMPs secreted by disc cells and involved in degradation of disc matrix rising as osmolarity falls. These results could explain in part the finding that MMP expression increases with degree of disc degeneration i.e. with loss of aggrecan and fall in extracellular osmolarity. These also suggested that a fall in osmolarity could induce a degenerative cascade with proteolytic digestion of aggrecan leading to a fall in osmolarity and hence a further increase in proteinase expression and matrix degradation.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:547482 |
| Date | January 2011 |
| Creators | Cui, Ying |
| Contributors | Urban, Jill |
| Publisher | University of Oxford |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://ora.ox.ac.uk/objects/uuid:90c2dec5-7c66-4fd8-9f45-55f8984d41e0 |
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