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Enhancement of a bone substitute by chemical pre-treatment to mobilize growth factor stores

Thomas, Matthew Bernard Miles 2012. Enhancement of a bone substitute by chemical pre-treatment to mobilize growth factor stores. MPhil Thesis, Cardiff University.
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Abstract

Growth factors are extracellular molecules that have several regulatory roles during inflammation and bone regeneration. Bio-Oss® granules and Bio-Oss® Collagen are xenografts used in the management of periodontal and periimplant defects, in addition to a variety of other clinical indications. The aims of this thesis were to investigate if Bio-Oss® contains residual growth factors that can be released by chemical pre-treatment. It also aimed to investigate if commercial Bio-Oss® granules or its extracted components alter cellular behaviour. Bio-Oss® collagen and Bio-Oss® granules underwent extraction with either ethylenediaminetetraacetic acid, calcium hydroxide or hydrochloric acid,followed by guanidinium chloride and tris(hydroxymethyl)aminomethane. Extracted proteins were separated using sodium dodecyl sulphate - polyacrylamide gel electrophoresis and stained with silver stain. Stained protein bands were consistently demonstrated in Bio-Oss® extracts. Immunoreactivity was demonstrated using antibodies to TGF-β1 and the Western blot technique. In order to investigate cellular behaviour, bone marrow stromal cells were harvested form Wistar rats prior to being cultured using mineralising media in the presence of Bio-Oss® granules and Bio-Oss® Collagen. Commercial Bio- Oss® in both forms was unable to support cell growth when seeded directly on their surfaces. In contrast, when cells were cultured with media supplemented with the extracted components of Bio-Oss® granules, cell behaviour was positively affected. Cell expansion was increased following exposure to DBBM extracts as measured with the MTS assay. Importantly, polymerase chain reactions revealed enhanced temporal expression of bone sialoprotein in cells cultured in the presence of DBBM extracted components. ii The detection of residual protein within Bio-Oss® granules in particular may be of clinical significance. The protein may be in the form of a complex of TGF-β1, a key growth factor, which may be osteoinductive. Alteration of gene expression profiles may suggest that faster bone regeneration is possible clinically if Bio-Oss® is chemically pre-treated to solubilise growth factors.

Item Type: Thesis (MPhil)
Status: Unpublished
Schools: Dentistry
Subjects: Q Science > Q Science (General)
R Medicine > RK Dentistry
Uncontrolled Keywords: bone substitute; growth factor; cell culture
Date of First Compliant Deposit: 30 March 2016
Last Modified: 24 Jul 2018 09:41
URI: http://orca.cf.ac.uk/id/eprint/39358

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