My research interests include developing an in vitro model to investigate the structural organisation of extracellular matrix (ECM) produced by cultured cells. This model has been used to compare the matrix produced by osteoblasts in culture to that in native bone using solid state NMR and scanning electron microscopy in collaboration with the Department of Chemistry and the Multi-Imaging Centre. We have developed a method for enriching the ECM with isotopes containing an asymmetric nuclear spin by culturing osteoblasts in the presence of the amino acids proline and glycine that have been labelled with the non-zero spin isotopes 13C and 15N. This enhances the signals for proline, hydroxyproline and glycine in type-1 collagen, the most abundant protein in the ECM produced by osteoblasts and allows us to compare these signals between ECM produced by cells in culture and in bone. I have also been using matrix labelling techniques to investigate the glycation of type I collagen in ECM formed by osteoblasts on exposure to high concentrations of glucose, which plays a role in diabetic bone pathology. This work has been combined with biochemical and molecular analysis of cells grown in high glucose medium and its effects on markers of oxidative stress.
Amgarten B, Rajan R, Martínez-Sáez N, Oliveira BL, Albuquerque IS, Brooks RA, Reid DG, Duer MJ, Bernardes GJ. (2015) Collagen labelling with an azide-proline chemical reporter in live cells. Chem Commun (Camb). 51(25):5250-2.
Wong VW, Reid DG, Chow WY, Rajan R, Green M, Brooks RA, Duer MJ. (2015) Preparation of highly and generally enriched mammalian tissues for solid state NMR. J Biomol NMR. 63(2):119-23.
Chow WY, Rajan R, Muller KH, Reid DG, Skepper JN, Wong WC, Brooks RA, Green M, Bihan D, Farndale RW, Slatter DW, Shanahan CM, Duer MJ (2014) NMR Spectroscopy of native and in vitro tissues implicates PolyADP Ribose in biomineralization. Science 344(6185), 742-6
Areas of expertise
cell culture, primary cell culture, molecular cell biology, immunofluorescence, osteoblast differentiation, extracellular matrix biology, NMR spectroscopy, solid state NMR, PCR, scanning electron microscopy, stem cell differentiation, cell isolation