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Title: Protein and cell interaction with bone substitute materials
Author: McCreary, Dara
ISNI:       0000 0004 8508 3887
Awarding Body: UCL (University College London)
Current Institution: University College London (University of London)
Date of Award: 2020
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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.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available