Dynamic Light Scattering Study of Hydroxyapatite Formation: Effect of Osteopontin and Bone Sialoprotein

.......................................................................................................................................... ii Acknowledgments.......................................................................................................................... iii List of Tables ................................................................................................................................. vi List of Figures ............................................................................................................................... vii Chapter 1: Introduction and Literature Review .............................................................................. 1 1.1 Biomineralization ............................................................................................................. 1 1.2 Crystal Nucleation and Growth ........................................................................................ 2 1.2.1 Nucleation ................................................................................................................. 2 1.2.2 Heterogeneous Matrix-Mediated Nucleation ............................................................ 5 1.2.3 Growth ...................................................................................................................... 6 1.3 Proteins ............................................................................................................................. 8 1.3.1 Osteopontin ............................................................................................................... 9 1.3.2 Bone sialoportein .................................................................................................... 12 1.4 Methods for Characterization of Protein Modulation of Mineralization ....................... 14 1.4.1 Steady State ............................................................................................................. 14 1.4.2 Constant Composition Method ............................................................................... 16 1.4.3 Dynamic Light Scattering ....................................................................................... 17 1.5 Purpose of this Thesis .................................................................................................... 19 Chapter 2: Methods ....................................................................................................................... 21 2.1 Dynamic Light Scattering .............................................................................................. 21 2.1.1 Theory of Dynamic Light Scattering ...................................................................... 21 2.1.2 Dynamic Light Scattering Experiment ................................................................... 25 2.1.3 Data Analysis .......................................................................................................... 26 2.2 Identification of Precipitates .......................................................................................... 28 2.2.1 X-ray crystallography ............................................................................................. 28 2.2.2 X-ray diffraction from a crystal: Bragg’s Law ....................................................... 28 2.2.3 Crystal Preparation.................................................................................................. 29