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http://eprints.org/relation/isVolatileVersionOf http://etheses.bham.ac.uk/id/document/3361 http://eprints.org/relation/ispreviewThumbnailVersionOf http://etheses.bham.ac.uk/id/document/3361 http://eprints.org/relation/isVersionOf http://etheses.bham.ac.uk/id/document/3361 archive 4031 disk0/00/00/13/83 2011-02-28 14:15:34 2019-01-23 09:54:03 2011-07-19 16:34:14 thesis show 0 Hunt Nicola Claire Grover Liam Shelton Richard M. University of Birmingham unpub TP 10col_ephy There is a clinical need for effective alternative skin replacements to autografts, allografts and xenografts. In this thesis a bi-layer skin graft was developed by encapsulation of fibroblasts in calcium-alginate hydrogel and culture of keratinocytes on the surface. Initially, the use of 5% and 2% w/v alginate hydrogels were investigated. Both scaffolds maintained fibroblast viability for at least 150 days encapsulation and caused reversible mitotic and catabolic inhibition, as assessed by fluorescent staining, immunochemistry and the thiazolyl blue assay. Sustained expression of angiogenic factors such as vascular endothelial growth factor, interleukin 6 and nerve growth factor were seen by fibroblasts encapsulated in both scaffolds, by reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay. Histological staining demonstrated that following degradation of the scaffolds, fibroblasts secreted ECM to facilitate dermal repair. Comparison of degradation of the scaffolds over time by measuring release of calcium, and changes in rheological properties, morphology and mass, indicated that 5% w/v alginate hydrogel degraded more slowly and was preferable to 2% w/v alginate hydrogel. Fibroblasts encapsulated in 5% w/v alginate hydrogel were shown to express keratinocyte growth factor by RT-PCR, to support keratinocyte proliferation and differentiation, and keratinocytes cultured on the 5% w/v alginate hydrogel surface were seen to form multi-layered epidermal structures by histology, immunostaining and RT-PCR. 2010-07 graduation eng This unpublished thesis/dissertation is copyright of the author and/or third parties. The intellectual property rights of the author or third parties in respect of this work are as defined by The Copyright Designs and Patents Act 1988 or as modified by any successor legislation. Any use made of information contained in this thesis/dissertation must be in accordance with that legislation and must be properly acknowledged. Further distribution or reproduction in any format is prohibited without the permission of the copyright holder. public 195 University of Birmingham School of Chemical Engineering d_ph european_commission 2010-07 graduation