Bibliography: leaves 148-160. / xix, 160, [40] leaves, [19] leaves of plates : ill. ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Investigates the process of neovascularization of intervertebral disc components, with ageing and in disease. Establishes the extent of end plate vascularization in the lumbar discs of young normal sheep, and demonstrates that some components of the disc undergo neovascularization in response to physical trauma, possibly as a part of tissue repair response. / Thesis (Ph.D.)--University of Adelaide, Dept. of Pathology, 1996?

Thesis (M.Sc.). / Written for the Dept. of Pharmacology and Therapeutics. Title from title page of PDF (viewed 2009/06/29). Includes bibliographical references.

published_or_final_version / abstract / Orthopaedics and Traumatology / Doctoral / Doctor of Philosophy

published_or_final_version / Biochemistry / Master / Master of Philosophy

Continuous monitoring during tissue culture is important for the success of engineered tissue development. It is also challenging due to lack of suitable established monitoring techniques. In this study, microdialysis, a sampling technique for measuring the unbound solute concentrations in the tissues and organs of the living body, was adopted to monitor functional tissue growth in a bioreactor with explanted bovine caudal intervertebral discs (IVD) as the test tissue. Apart from cell metabolic activities, cell and tissue biological functions were investigated for the development of microdialysis for monitoring purposes. Methodologies of microdialysis with large pore size membrane probes for sampling macromolecular bio-functional markers were established. The effects of pumping methods, including 'push', 'pull' or 'push-and-pull', and the effect of the resulting transmembrane pressure on the fluid balance, and the relative recovery of small molecules and of macromolecules (proteins) were experimentally studied. The validity of the internal reference in-situ calibration was examined in detail. It was concluded that a push-and-pull system was the only effective method to eliminate fluid loss or gain. The relative recovery of small solutes was hardly affected by the applied pumping methods; however the relative recovery of macromolecules was significantly influenced by them. The in situ calibration technique using Phenol Red can provide reliable results for small molecules including glucose and lactic acid. Using lOkDa and 70kDa fluorescent dextrans as the internal standard for in situ calibration of large molecules of similar size, it was found that the pull pump system did not work well but that the push-and-pull pumping method did work well. A novel bioreactor system for in vitro IVD culture with static load and microdialysis monitoring was developed. Explanted IVDs were cultured under three different loads for up to 7 days. A single microdialysis probe with 3000 kDa membrane was inserted into each of the IVDs at a defined location. The in situ calibration technique was proved valid in the experiments and membrane fouling was not significant. The tissue metabolism and extracellular matrix turnover during 7 day culture were continuously monitored to investigate the effect of different loads. Microdialysis proved to be a feasible and efficient method for multi-parameter monitoring of tissue culture. Substantial effort was directed towards the identification of functional macromolecular markers in conjunction with microdialysis sampling. Amongst several proteins sampled, chitinase-3-like protein 1 (CHI3L1), a major soluble protein secreted by cultured IVD cells in alginate beads and by cultured IVD explants was identified following its successful isolation. Then it was established as a suitable functional marker. The effect of physico-chemical and mechanical stimuli (e.g. osmolarity, pH, oxygen tension and mechanical load) on secretion of CHI3L1 by cultured IVD cells and chondrocytes in alginate beads and by cultured IVD explant were investigated. CHI3L1 release was sensitive to physico-chemical stimulation. The production of CHI3L1 was directly correlated with the cell metabolism and this could be readily monitored with microdialysis.

No description available.

Bibliography: leaves 107-109.

Thesis (Ph. D.)--University of Hong Kong, 2010. / Includes bibliographical references (leaves 149-163). Also available online.

Thesis (Ph. D.)--University of Hong Kong, 2010. / Includes bibliographical references (leaves 149-163). Also available in print.

published_or_final_version / Orthopaedics and Traumatology / Doctoral / Doctor of Philosophy