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Use of comparative proteomics to study a novel osteogenic nanotopography

Kantawong, Fahsai (2009) Use of comparative proteomics to study a novel osteogenic nanotopography. PhD thesis, University of Glasgow.

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Abstract

The principal aim of this thesis was to investigate the ability of surface topography in inducing bone cell differentiation for biomedical purposes. In orthopedic research, regeneration of bone defects can be performed in vitro using biomaterials. Third generation biomaterials aim not only to support tissue (first generation) and not only to be ‘bioactive’ (second generation), but to stimulate specific, known and desirable responses at the molecular level. Nanoscale topography offers a possible route to the development of third generation biomaterials. Two-dimensional fluorescence difference gel electrophoresis (2D DIGE) is a new method for assessing protein expression strategies and here, a micro-grooved topography was used as a model for protocol optimization. The protocol was successfully developed and proved that 2D DIGE can be used as a powerful tool in the evaluation of biomaterial can direct cell behavior and cell fate. Next, the refined protocol was applied to the evaluation of the novel nanotopographic features; near-square nanopits (120 nm diameter, 100 nm depth with the pitch between the pits was set to an average of 300 nm with a ± 50 nm error). Protein expression profiles indicated that ERK1/2 might play part in cell proliferation and cell differentiation. However, to make a clear conclusion about molecular signalling, the study of sub-cellular proteome is needed in the future work. Additionally, the use of another comparative proteomic technique; dimethyl labelling, implicated the possibility of sub-population differentiation, i.e. the formation of multiple cell types that could be advantageous in tissue engineering of complex organs. Furthermore, the application of fluid-flow bioreactors was shown to enhance the growth rate and possibly increased differentiation of cells cultured on nanotopographical features.

Item Type: Thesis (PhD)
Qualification Level: Doctoral
Additional Information: 1. KANTAWONG, F., BURCHMORE, R., WILKINSON, C. D., OREFFO, R. O. & DALBY, M. J. (2009) Differential in-gel electrophoresis (DIGE) analysis of human bone marrow osteoprogenitor cell contact guidance. Acta Biomater, 5, 1137-46. 2. KANTAWONG, F., BURCHMORE, R., GADEGAARD, N., OREFFO, R. O. & DALBY, M. J. (2008) Proteomic analysis of human osteoprogenitor response to disordered nanotopography. J R Soc Interface. 3. KANTAWONG, F., BURGESS, K. E., JAYAWARDENA, K., HART, A., BURCHMORE, R. J., GADEGAARD, N., OREFFO, R. O. & DALBY, M. J. (2009) Whole proteome analysis of osteoprogenitor differentiation induced by disordered nanotopography and mediated by ERK signalling. Biomaterials.
Keywords: Nanotopography, Proteomics, Osteoprogenitor cell
Subjects: Q Science > QR Microbiology
Q Science > Q Science (General)
Colleges/Schools: College of Medical Veterinary and Life Sciences > Institute of Molecular Cell and Systems Biology
Supervisor's Name: Dalby, Dr. Matthew and Burchmore, Dr. Richard
Date of Award: 2009
Depositing User: Miss Fahsai Kantawong
Unique ID: glathesis:2009-919
Copyright: Copyright of this thesis is held by the author.
Date Deposited: 15 Jul 2009
Last Modified: 10 Dec 2012 13:28
URI: http://theses.gla.ac.uk/id/eprint/919

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