HISTOMORPHOMETRY OF THE HUMAN RIB CORTEX IN METHAMPHETAMINE USERS by Robert Chancey Karinen A thesis submitted in partial fulfillment of the requirements for the degree of Master of Arts in Anthropology Boise State University December 2009 BOISE STATE UNIVERSITY GRADUATE COLLEGE DEFENSE COMMITTEE AND FINAL READING APPROVALS of the thesis submitted by Robert Chancey Karinen Thesis Title: Histomorphometry of the human rib cortex in methamphetamine users Date of Final Oral Examination: 14 October 2009 The following individuals read and discussed the thesis submitted by student Robert Chancey Karinen, and they evaluated his presentation and response to questions during the final oral examination. They found that the student passed the final oral examination. Margaret Streeter, Ph.D. Chair, Supervisory Committee Mark Plew, Ph.D. Member, Supervisory Committee Christopher Hill, Ph.D. Member, Supervisory Committee The final reading approval of the thesis was granted by Margaret Streeter, Ph.D., Chair of the Supervisory Committee. The thesis was approved for the Graduate College by John R. Pelton, Ph.D., Dean of the Graduate College. ACKNOWLEDGEMENTS Dr. Margaret Streeter- for serving as my advisor and chair of my committee. For introducing me to bone histology. For the countless hours of advice and education. I was truly lucky to have a advisor that is whole heartedly committed to the education of her students. Dr. Mark Plew- for serving on my committee and providing support and encouragement throughout my graduate education. Dr. Christopher Hill- for serving on my committee. The education in statistics provided by you is what gives my research meaning. Ada County Coroners Office- For keeping me employed during this process and allowing me a source from which to collect data. To my fellow students from which I have learned from and who have made this experience all the more rewarding. To my wife Jenny who has always supported me in whatever endeavor I undertake. Thank you for everything you do for our family. I look forward to being able to return to our life together. To the decedents included in the study. i ABSTRACT Studies have demonstrated lifestyle choices such as poor diet, insufficient exercise and substance abuse can negatively affect bone health. The purpose of this research is to determine if the tissue pathology associated with long term methamphetamine use is a localized response to poor dental hygiene or an indication of a more systemic response that is discernable in the bone microstructure. A comparison of the rib cortical bone microstructure between males that were known to be methamphetamine abusers (N=18) and individuals who did not abuse the drug was undertaken (N=19). Histomorphometric variables calculated in this analysis included mean osteon size, osteon population density (OPD) and cortical area measurement. OPD was found to vary significantly between study and control populations. This study demonstrates that methamphetamine abuse is associated with changes in OPD. ii TABLE OF CONTENTS ACKNOWLEDGMENTS .......................................................................................... i ABSTRACT ............................................................................................................... ii LIST OF TABLES ...................................................................................................... vi LIST OF FIGURES .................................................................................................... vii LIST OF CHARTS AND GRAPHS ........................................................................... viii CHAPTER ONE: INTRODUCTION........................................................................... 1 Background ..................................................................................................... 4 Bone Composition ............................................................................................ 4 Functions of Bone ........................................................................................... 4 Bone Cells ....................................................................................................... 5 Bone Mass and Bone Density ......................................................................... 6 Bone Growth, Modeling and Remodeling ...................................................... 6 Modeling and Growth ......................................................................... 6 Remodeling ......................................................................................... 7 The Basic Multicellular Unit .............................................................. 9 Growth of Ribs ................................................................................................ 10 Anatomy of a Rib ............................................................................................ 11 Interstitial Bone and Osteons .......................................................................... 12 Cortical Area ................................................................................................... 15 iii Mean Osteon Size ........................................................................................... 16 Osteon Population Density ............................................................................. 17 Histological Estimation of Age ....................................................................... 18 Use of Rib Microstructure in Age Estimation ................................................ 19 Variables Affecting Bone Remodeling ........................................................... 20 Age ...................................................................................................... 20 Sex ....................................................................................................... 21 Ancestry .............................................................................................. 22 Mechanical Stress ............................................................................... 23 Dietary Influences ............................................................................... 24 Pathology ............................................................................................ 25 Drugs ................................................................................................... 26 Methamphetamine ........................................................................................... 28 Methamphetamine and Bone Quality ................................................. 33 CHAPTER TWO: MATERIALS AND METHODS ................................................. 34 Methamphetamine User Sample ..................................................................... 34 Comparative Sample ....................................................................................... 34 CHAPTER THREE: METHODS ............................................................................... 37 Collection and Preparation of Materials ......................................................... 37 Preparation of Slides ....................................................................................... 37 Microscopic Analysis ...................................................................................... 37 iv Histomorphometry .......................................................................................... 38 Mean Osteon Area .............................................................................. 38 Cortical Area Calculations .................................................................. 39 OPD ..................................................................................................... 40 Histological Age Estimation ............................................................... 41 CHAPTER FOUR: RESULTS ................................................................................... 42 OPD ............................................................................................................... 42 Mean Osteon Size ........................................................................................... 45 Mean Cortical Area ......................................................................................... 46 CHAPTER 5: DISCUSSSION ..................................................................................... 48 OPD ............................................................................................................... 48 Implications for Histological Age Estimations ................................... 48 Mean Osteon Size ........................................................................................... 49 Cortical Area ................................................................................................... 50 CHAPTER 6: CONCLUSIONS ................................................................................. 51 REFERENCES ........................................................................................................... 55 APPENDIX ............................................................................................................... 67 Waiver Form v LIST OF TABLES Table 5.1 Summary Statistics Methamphetamine Users vs. Non-Users............... 42 Table 5.2 Basic Statistics OPD Users vs. Non-Users ........................................... 43 Table 5.3 Mean Osteon Size Users vs. Non-Users ............................................... 45 Table 5.4 Basic Statistics Cortical Area Users vs. Non-Users.............................