McCreary, Dara;
(2020)
Protein and Cell Interaction with Bone Substitute Materials.
Doctoral thesis (Ph.D), UCL (University College London).
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McCreary PhD Thesis Protein and Cell interactions with Bone substitute materials.pdf Download (4MB) | Preview |
Abstract
Protein attachment to bone graft substitutes (BGS) is believed to influence the cell and tissue response to these materials. This thesis aimed was to investigate protein attachment to the surface of five commercial BGSs and to determine if this affected cellular response and osteoinduction in vitro. I hypothesised that differences in BGS composition would encourage different proteins to adhere and thereby influence subsequent cellular behaviour. Furthermore, I hypothesised that when selected proteins were coated onto the BGS surface, they would enhance cell proliferation and osteogenic differentiation. No difference in mesenchymal stem cell differentiation was found on the different BGSs, but there was a significant increase in cell viability and proliferation on Inductigraft™ when compared with ApaPore™, Actifuse™, β-TCP™ and Orthoss® (p=0.001). Sintered discs of hydroxyapatite (HA) (ApaPore™) and silicate-substitute HA (SiHA) (Actifuse™ and Inductigraft™) were created to remove the influence of BGS morphology on cell behaviour and used to investigate the effect of silicon substitution on cell proliferation and osteogenic differentiation. The incorporation of Si significantly increased metabolic activity (p=0.001). Protein adhesion was measured following incubation with foetal calf serum for 30s, 1hr, 24hrs and 72hrs using SDS Page and micro BCA assays. Higher protein attachment was observed on Orthoss® compared to other BGSs. Mass spectrometry identified 106 adhered proteins and showed differences in the protein attachment profiles on the different BGSs. To promote osteoinduction, proteins identified from the adhesion study, that could potentially affect osteoinduction, were selected (fibronectin, collagen type I alpha) and coated onto sintered HA. These proteins significantly increased cell metabolic activity (p=0.001; p=0.003). My study has shown that protein adhesion is essential, and it may be possible to select and coat proteins onto the BGS that will influence subsequent bone formation
| Type: | Thesis (Doctoral) |
|---|---|
| Qualification: | Ph.D |
| Title: | Protein and Cell Interaction with Bone Substitute Materials |
| Event: | UCL |
| Open access status: | An open access version is available from UCL Discovery |
| Language: | English |
| Additional information: | Copyright © The Author 2020. Original content in this thesis is licensed under the terms of the Creative Commons Attribution 4.0 International (CC BY 4.0) Licence (https://creativecommons.org/licenses/by/4.0/). Any third-party copyright material present remains the property of its respective owner(s) and is licensed under its existing terms. Access may initially be restricted at the author’s request. |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health > Infection, Immunity and Inflammation Dept |
| URI: | https://discovery-pp.ucl.ac.uk/id/eprint/10089587 |
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