Evaluation of novel salivary peptides and a new application method for prevention and treatment of oral disease

.............................................................................................................................. ii Co-Authorship ................................................................................................................... iii Acknowledgments ............................................................................................................. iv Table of Contents ................................................................................................................ v List of Tables ................................................................................................................... viii List of Figures .................................................................................................................... ix List of Abbreviations .......................................................................................................... x Chapter 1 ............................................................................................................................. 1 1.1 Introduction (Problem statement in saliva biology) ............................................... 1 1.2 References ............................................................................................................... 7 Chapter 2 ........................................................................................................................... 11 2 Protein adsorption on the enamel and their biological function .................................. 11 2.1 Introduction ........................................................................................................... 11 2.2 AEP Composition ................................................................................................. 13 2.2.1 Amylase .................................................................................................... 14 2.2.2 Histatins .................................................................................................... 15 2.2.3 Mucin ........................................................................................................ 17 2.2.4 Proline-rich proteins (PRPs) ..................................................................... 19 2.2.5 Statherin .................................................................................................... 21 2.2.6 Lysozyme .................................................................................................. 22 2.3 References ............................................................................................................. 24 Chapter 3 ........................................................................................................................... 36 3 Duplication and Hybridization of Protein Functional Domains for Oral Homeostasis 36 vi 3.1 Introduction ........................................................................................................... 36 3.2 Materials and Methods ......................................................................................... 38 3.2.1 Protein and peptides characterization ....................................................... 38 3.2.2 Calculation of protein and peptide isoelectric points ............................... 38 3.2.3 Enamel demineralization assay ................................................................. 39 3.2.4 Calcium and phosphate analyses .............................................................. 39 3.2.5 Candida albicans killing assay ................................................................. 40 3.2.6 Statistical analyses .................................................................................... 40 3.3 Results ................................................................................................................... 41 3.4 Discussion ............................................................................................................. 46 3.5 References ............................................................................................................. 49 4 The effect of pH on killing properties of chitosan nanoparticles against Candida albicans ........................................................................................................................ 52 4.1 Introduction ........................................................................................................... 52 4.2 Material and methods ........................................................................................... 54 4.2.1 Chitosan nanoparticles’ construction ........................................................ 54 4.2.2 Candida albicans killing assay .................................................................. 55 4.2.3 Evaluation of cell viability ........................................................................ 55 4.2.4 Chitosan nanoparticles suspension ........................................................... 55 4.3 Results ................................................................................................................... 56 4.3.1 Chitosan nanoparticles possess a more effective antifungal activity compared to plain chitosan ....................................................................... 56 4.3.2 The Killing effect of chitosan nanoparticles made with different initial concentrations of chitosan proved to be significantly different ................ 58 4.3.3 Chitosan nanoparticles show a most efficient killing effect in pH 4 ........ 60 4.4 Discussion ............................................................................................................. 63 4.5 References ............................................................................................................. 65