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This dissertation, Fabrication of Multi-component Tissue for Intervertebral Disc Tissue Engineering by Tsz-kit, Chik, 戚子傑, was obtained from The University of Hong Kong (Pokfulam, Hong Kong) and is being sold pursuant to Creative Commons: Attribution 3.0 Hong Kong License. The content of this dissertation has not been altered in any way. We have altered the formatting in order to facilitate the ease of printing and reading of the dissertation. All rights not granted by the above license are retained by the author. Abstract: Intervertebral disc tissue engineering is challenging because it involves the integration of multiple tissues with distinct structures and compositions such as lamellar annulus fibrosus, gel?like nucleus pulposus and cartilage endplate. Each of them has different compositions and different structures. It is hypothesized that integration of tissues can be enhanced with appropriate mechanical and biological stimuli. Meanwhile, effect of torsional stimulus on cell re?orientation in mesenchymal stem cell?collagen tubular constructs is investigated in this study. Furthermore, it is proposed that these findings can be used to fabricate a multicomponent unit for intervertebral disc tissue engineering. It has been demonstrated that mechanical and biological stimuli can stabilize the interface between osteogenic and chondrogenic differentiated constructs with enhanced ultimate tensile stress while the phenotype of osteogenic and chondrogenic differentiated constructs were maintained. Scanning electronic microscopic images have shown aligned collagen fibrils and presence of calcium at the interface, indicating the possibility of the formation of a calcified zone. In addition, it is proven that torsional stimulus triggered re?orientation of mesenchymal stem cells in collagen lamellae towards a preferred angle. Cell alignments were confirmed by using a MatLab?based program to analyze the actin filament and the cell alignment via Phalloidin and Hematoxylin staining, respectively. Cells and actin filaments were inclined around 30o from the vertical axis, while cells and filaments in the control group (static loading) aligned along the vertical axis. Furthermore, a double?layers bioengineered unit was fabricated, with intact osteogenic differentiated parts at both ends. Comparatively higher cell density was observed at the interface between layers, demonstrating the interactions between layers, while the phenotype of each part was maintained in 14 days culture. This study concludes that a multi?components bioengineered unit with preferred cell alignments can be fabricated. This provides new insights to future development of bioengineered spinal motion segment for treating late stage disc degeneration. DOI: 10.5353/th_b4784944 Subjects: Intervertebral disk prosthesesTissue engineering - Materials

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