Thesis
Engineering of artificial bone tissues
- Creator
- Rights statement
- Awarding institution
- University of Strathclyde
- Date of award
- 2016
- Thesis identifier
- T14272
- Person Identifier (Local)
- 201358340
- Qualification Level
- Qualification Name
- Department, School or Faculty
- Abstract
- Bone defects caused by trauma and pathological and physiological bone resorption denote a major challenge and have become a global health problem. Decellularised bone has been proposed in bone tissue engineering studies as a donor scaffold for bone implantation into patients. The aim of a decellularisation approach is to efficiently eliminate all donor cellular material while minimising any adverse effect on the composition, mechanical integrity, and biological activity of the remaining extracellular matrix. In fact, the major aim of this study is to develop an efficient protocol to decellularise and recellularise the xenogenic bone so that it can be used for bone regeneration applications. The effectiveness of donor cell removal from a tissue is influenced by the origin of the tissue and also the use of specific physical, chemical, and enzymatic methods. In this study, the success of a bovine bone decellularisation using 10 mM Tris, 1 mM EDTA, 0.1 % (v/v) Triton X-100 and 0.5 % (w/v) Trypsin has been established through analytical DNA assays. This study was further extended to measure the mechanical properties of bone before and after decellularisation. Compression mechanical testing has been applied. Mechanical testing of square-shaped specimens loaded in compression was accomplished using the Bose ElectroForce 3200 to ensure maintenance of mechanical integrity. Likewise, the pore sizes of bone samples were measured before and after decellularisation using the PoreMaster-60 to ensure the scaffolds have an ideal diameter for cells to grow on/into after the decellularisation process. Further to establish the recellularisation protocol, reseeding using HOS cells was allowed to progress for up to 5 weeks in the presence of an adherent and a non-adherent surface. Several test parameters such as alkaline phosphatase (ALP) activity and DAPI staining for DNA presence were measured. The culture samples were also compared in static and dynamic culture conditions. After 3 weeks of in vitro culture, the reseeded cells and materials in adherence and non-adherence conditions were measured for the MTT reduction, mechanical testing, pore diameter, and human osteocalcin syhthesis by an ELISA kit. The adherence and proliferation within the scaffolds were then confirmed by the SEM after 21 days post reseeding. The effects of vitamin D3 on the cell growth were also carried out. Overall, the findings presented that the decellularised bovine bone scaffolds are capable of supporting cell adherence and proliferation. Moreover, some test parameters after recellularisation have shown the potential use of this bone scaffolds in vivo. Taken together, the studies carried out in this project show that this bovine donor bone scaffold could be used to repair bone defects in recipient patients.
- Advisor / supervisor
- Grant, Helen
- Riches, Philip
- Resource Type
- DOI
- Embargo Note
关系
项目
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File | 2023-03-07 | 非公开 |