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Title: A study of human mesenchymal stem cells, human primary osteoblasts and osteoblast-like cells using Raman spectroscopy
Author: McManus, Lindsay L.
ISNI:       0000 0004 2717 6508
Awarding Body: University of Ulster
Current Institution: Ulster University
Date of Award: 2011
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Raman spectroscopy is a vibrational spectroscopy technique that provides a global biochemical signature and has been shown to have utility in the analysis of biological cells for bone tissue engineering applications. Traditionally, sample analysis in this field employs destructive biological methods that require the use of biomarkers, however, Raman has since become an essential tool in various areas of bio-industry and by incorporating the technique into biological laboratories these perturbing methodologies are no longer the only means of analysis. Therefore the focus of this study was to investigate the capability of Raman spectroscopy as a tool for the in vitro characterisation of the sub-cellular composition and osteogenic potential of human mesenchymal stem cells. As with most biological samples a high degree of heterogeneity is often found, therefore in order to extract the desired information from the biological studies multivariate analysis tools were utilised. The reliability and consistency of the vibrational analysis was confirmed by means of comparison with current gold-standard techniques such as, alizarin red staining, real-time polymerase chain reaction and immunocytochemistry. A further novelty was introduced with the use of Raman spectroscopy to determine the suitability of the U20S osteoblast-like cell line for use as a model for human primary osteoblasts with emphasis on the ability of these cell types to replicate their tissue of origin. Investigation of the U20S osteoblast-like cell line provided evidence of dense multilayered mineralised regions that corresponded more closely to native bone, which has not been previously reported on. This finding contradicts previous reports on U20S osteoblast-like cells which have consistently been described as non-osteoinductive. When Raman spectroscopy is coupled with biological and multivariate analyses techniques, it shows further novelty when employed to monitor mineralisation of human mesenchymal stem cells, human primary osteoblasts and osteoblast-like cells. This body of work culminates the success of a truly multidisciplinary approach and provides the potential for further work on bone cell analysis and the applications of spectroscopy for biological studies and bone tissue engineering applications.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available