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The Influence of Salivary Statherin, Histatin-1 and Their 21 N-Terminal Peptides Individually and When in Combination on The

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The Influence of Salivary Statherin, Histatin-1 and Their 21 N-Terminal Peptides Individually and When in Combination on The The Influence of Salivary Statherin, Histatin-1 and Their 21 N-terminal Peptides Individually and When in Combination on the Demineralisation of Hydroxyapatite and Enamel. The Effect of Peptides Adsorption, Aggregation, Surface Charge and Secondary Structure Huda Barak A Almandil BDS (KSU), MClinDent Paediatric Dentistry(QMUL) Supervised by Prof. Paul Anderson, Dr Maisoon Al-Jawad Thesis submitted in fulfilment of the requirements for the degree of Doctor of Philosophy in the University of London 2018 Dental Physical Sciences Unit, Institute of Dentistry, Barts and the London School of Medicine and Dentistry, Queen Mary University of London I Huda Barak Almandil, confirm that the research included within this thesis is my own work or that where it has been carried out in collaboration with, or supported by others, that this is duly acknowledged below, and my contribution indicated. Previously published material is also acknowledged below. I attest that I have exercised reasonable care to ensure that the work is original and does not to the best of my knowledge break any UK law, infringe any third party’s copyright or other Intellectual Property Right, or contain any confidential material. I accept that the College has the right to use plagiarism detection software to check the electronic version of the thesis. I confirm that this thesis has not been previously submitted for the award of a degree by this or any other university. The copyright of this thesis rests with the author and no quotation from it or information derived from it may be published without the prior written consent of the author. Signature: Huda Almandil Date:14/2/2018 i Table of Contents Figure Index ....................................................................................................... viii Table Index ........................................................................................................ xvi List of Abbreviations ........................................................................................ xvii Acknowledgements ......................................................................................... xviii Abstract ................................................................................................................ xix Chapter 1. General Introduction......................................................................... 1 Aims and Objectives ................................................................................ 2 Chapter 2. Enamel and Enamel Demineralisation .............................................. 5 Dental Hard Tissues ................................................................................ 5 Enamel .................................................................................................... 5 Enamel Mineral Composition ................................................................. 9 Enamel Caries ........................................................................................ 10 2.4.1 Histological Changes in Caries Enamel ............................................ 11 2.4.2 Dissolution Models .......................................................................... 13 Erosion .................................................................................................. 17 Hydroxyapatite as a Model System for Dental Enamel ........................ 20 Chapter 3. Saliva and Dental Health ................................................................. 22 Saliva ..................................................................................................... 22 Salivary Secretion .................................................................................. 23 General Function of Saliva .................................................................... 26 3.3.1 Food Taste, Clearance and Digestion .............................................. 26 3.3.2 Lubrication ....................................................................................... 27 3.3.3 Buffering .......................................................................................... 27 3.3.4 Antimicrobial Effects ........................................................................ 28 3.3.5 Preservation of Tooth Integrity ....................................................... 29 Acquired Enamel Pellicle ....................................................................... 30 Chapter 4. Salivary Statherin and Histatin ........................................................ 33 Salivary Human Statherin (STN) ............................................................ 33 Salivary Human Histatins ...................................................................... 38 Chapter 5. Salivary Proteins/Peptides and The Mineralised Tissue Affinity .... 42 Hydroxyapatite and Enamel as an Adsorbent Surface ......................... 42 ii Role of Amino Acids and Their Functional Groups in Peptide/Protein and HA Adsorption 43 Sequences and Conformation of Peptide/Protein Accountable for the Adsorption onto HA ...................................................................................................... 46 Thermodynamics of Peptide/Protein Adsorption to HA ....................... 48 Effect of Different Biological Molecules on Protein/Peptide Adsorption 49 Affinity of Salivary Proteins/Peptides for HA/Enamel .......................... 50 Salivary Proteins/Peptides and Enamel Homeostasis........................... 51 5.7.1 Buffering Capacity ............................................................................ 51 5.7.2 Inhibition of Calcium Phosphate Salts Precipitation........................ 52 5.7.3 Inhibition of Mineral Demineralisation ........................................... 52 TECHNIQUES USED IN THIS STUDY ......................................................... 54 Chapter 6. Scanning Microradiography (SMR) ................................................. 55 Scanning Microradiography (SMR) ....................................................... 55 6.1.1 Foundation of Electromagnetic Radiation ....................................... 55 6.1.2 X-ray Production .............................................................................. 56 6.1.3 X-ray Tube ........................................................................................ 57 6.1.4 Micro-focus Tubes ........................................................................... 58 6.1.5 Electron Impact X-ray Source .......................................................... 58 X-ray Interaction with Matter ............................................................... 59 6.2.1 Photoelectric Absorption ................................................................. 60 6.2.2 Compton Scattering ......................................................................... 60 6.2.3 Rayleigh Scattering .......................................................................... 61 6.2.4 Pair Scattering .................................................................................. 61 6.2.5 X-ray Attenuation: Beer’s law .......................................................... 61 X-ray Detection ..................................................................................... 63 6.3.1 Semiconductor Detectors ................................................................ 63 Multi-channel Analyser (MCA) .............................................................. 64 SMR Apparatus ..................................................................................... 65 6.5.1 X-ray Generator ............................................................................... 65 6.5.2 X-ray Detector .................................................................................. 66 6.5.3 SMR Stage ........................................................................................ 66 6.5.4 SMR Cell ........................................................................................... 66 iii Data Analysis at a Scanning Position .................................................... 67 Calculation of The Mineral Mass loss ................................................... 68 6.7.1 Percent Reduction in the Rate of Mineral Loss (PRRML) ................ 69 Responsiveness of SMR ........................................................................ 71 Advantages of SMR ............................................................................... 71 Chapter 7. Nano-Spectrophotometry ............................................................... 73 Introduction .......................................................................................... 73 Beer-Lambert Law ................................................................................. 74 Protein Absorption ................................................................................ 75 Advantages of Nano-Spectrophotometry ............................................. 76 Chapter 8. Dynamic Light Scattering (DLS) ....................................................... 77 Particle Size Measurement ................................................................... 77 Zeta Potential ........................................................................................ 79 Advantages of DLS ................................................................................ 79 Chapter 9. Circular Dichroism (CD) ................................................................... 81 Background ........................................................................................... 81 Theory ..................................................................................................
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