LYMPHOCYTE AND MACROPHAGE INTERACTIONS IN THE RESPONSE TO BIOMATERIAL SURFACES BY DAVID T. CHANG Submitted in partial fulfillment of the requirements For the degree of Doctor of Philosophy Dissertation Advisor: Dr. James M. Anderson Department of Biomedical Engineering CASE WESTERN RESERVE UNIVERSITY August, 2008 CASE WESTERN RESERVE UNIVERSITY SCHOOL OF GRADUATE STUDIES We hereby approve the thesis/dissertation of _____________________________________________________ candidate for the ______________________degree *. (signed)_______________________________________________ (chair of the committee) ________________________________________________ ________________________________________________ ________________________________________________ ________________________________________________ ________________________________________________ (date) _______________________ *We also certify that written approval has been obtained for any proprietary material contained therein. Copyright © 2008 by David T. Chang All rights reserved Table of Contents List of Tables .....................................................................................................................iii List of Figures..................................................................................................................... v Acknowledgements..........................................................................................................viii List of Abbreviations ......................................................................................................... ix Glossary ............................................................................................................................ xii Abstract............................................................................................................................. xv Chapter I. Introduction........................................................................................................1 Biological Response to Biomaterials.............................................................................1 Protein Adsorption and Complement.............................................................................5 Cellular Adhesion on Biomaterials................................................................................7 Macrophage Responses to Biomaterial Surfaces.........................................................10 Lymphocyte Responses to Biomaterials: Lymphocyte/Macrophage Interactions...................................................................................................................16 Inflammatory Mediators in Inflammation, Foreign Body Reaction, and Wound Healing ...........................................................................................................20 Mathematical Modeling...............................................................................................27 Model Biomaterial Surfaces ........................................................................................30 Significance..................................................................................................................45 Hypothesis and Specific Aims.....................................................................................46 References....................................................................................................................47 Chapter II. Lymphocyte/Macrophage Interactions: Biomaterial Surface- Dependent Cytokine, Chemokine, and Matrix Protein Production ...................................60 Abstract........................................................................................................................60 Introduction..................................................................................................................61 Materials and Methods.................................................................................................64 Results..........................................................................................................................69 Discussion....................................................................................................................81 References....................................................................................................................88 Chapter III. Paracrine and Juxtacrine Lymphocyte Enhancement of Adherent Macrophage and Foreign Body Giant Cell Activation ......................................................93 Abstract........................................................................................................................93 Introduction..................................................................................................................94 Materials and Methods.................................................................................................96 Results..........................................................................................................................99 Discussion..................................................................................................................106 i References..................................................................................................................111 Chapter IV. The Effect of Biomaterial Surface Chemistry on Adherent Lymphocyte Interactions with Macrophages and Foreign Body Giant Cells..................113 Abstract......................................................................................................................113 Introduction................................................................................................................114 Materials and Methods...............................................................................................117 Results........................................................................................................................120 Discussion..................................................................................................................131 References..................................................................................................................142 Chapter V. IFN-γ Production from Lymphocyte Interactions with Biomaterial- Adherent Macrophages and Foreign Body Giant Cells ...................................................147 Abstract......................................................................................................................147 Introduction................................................................................................................148 Materials and Methods...............................................................................................150 Results........................................................................................................................152 Discussion..................................................................................................................155 References..................................................................................................................170 Chapter VI. Dynamic Systems Model for Lymphocyte Interactions with Macrophages at Biomaterial Surfaces .............................................................................175 Introduction................................................................................................................175 Experimental Methods...............................................................................................178 Model Development...................................................................................................179 Results........................................................................................................................195 Discussion..................................................................................................................203 References..................................................................................................................212 Chapter VII. Conclusions and Future Directions............................................................216 Summary and Conclusions ........................................................................................216 Future Directions.......................................................................................................224 References..................................................................................................................228 Appendix I. Dynamic Systems Model Main Code .........................................................230 Appendix II. Dynamic Systems Model Output Function ...............................................246 Appendix III. Dynamic Systems Model Differential Equations Function .....................250 Works Cited .....................................................................................................................253 ii List of Tables Table 1.1: Adhesion Molecules 10 Table 1.2: T Lymphocyte Subpopulations and Functions 19 Table 1.3: Important Mediators Involved in Inflammation and Wound Healing 23 Table 1.4: Mechanical and Physical Properties of PET 32 Table 1.5: Composition and Contact Angles for PET-based Surfaces 44 Table 2.1: Minimum Detection Limit for the Utilized Protein Detection Techniques 68 Table 2.2: Signal Intensity Ratings and Material Variability Indices 72 Table 4.1: Adherent Lymphocyte Densities (cells/mm2) on Biomaterial Surfaces with Varying Monocyte Concentrations