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Effects of Ultraviolet Radiation Exposure on Oxidative Stress Markers on the Human Ocular Surface

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Effects of Ultraviolet Radiation Exposure on Oxidative Stress Markers on the Human Ocular Surface Effects of Ultraviolet Radiation Exposure on Oxidative Stress Markers on the Human Ocular Surface DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By Kristina Marie Haworth, OD, MS Graduate Program in Vision Science The Ohio State University 2014 Dissertation Committee: Heather Chandler, PhD, Advisor Donald Mutti, OD, PhD Karla Zadnik, OD, PhD Tatiana Oberyszyn, PhD Copyright by Kristina Marie Haworth 2014 Abstract Purpose: To evaluate feasibility, repeatability, and relationships between measures of ocular sun exposure, conjunctival ultraviolet autofluorescence (UVAF), hexanoyl-lysine (HEL), and 8-hydroxy-2’-deoxyguanosine (8OHdG) for in vivo human tear and conjunctival cell sample types. Methods: Fifty healthy volunteers were seen for 2 visits 14±2 days apart. Ocular sun exposure was estimated by questionnaires quantifying time outdoors and ocular protection habits. Conjunctival UVAF images were obtained using a Nikon D7000 camera system adapted with filters to photograph only UV excited visible light. Collection of tear samples was by glass microcapillary tube; conjunctival cells by cytology brush. Image analysis was accomplished using ImageJ image analysis software. HEL and 8OHdG ELISAs were conducted for both tear and conjunctival cell samples. Normality of distributions for each outcome measure was evaluated (Shapiro-Wilk); data was transformed to approximate normal distributions. Dichotomous outcome variables for occupation, contact lens wear, conjunctival UVAF, and ocular sun exposure were evaluated. Furthermore, estimates of repeatability for ocular sun exposure, UVAF and the ELISAs were evaluated using Bland-Altman plots with mean bias and 95% limits of agreement calculated. Linear or logistic regression was used to evaluate for relationships of continuous and dichotomous outcome variables respectively. Multiple linear regression models were built using a forward stepwise approach. ii Results: Descriptive statistics were generated for outcome measures of ocular sun exposure, conjunctival UVAF, HEL expression in tears and cells, and 8OHdG expression in tears. Bland-Altman plots produced demonstrated repeatability for each outcome measure and for the ELISA assays. Non-normally distributed raw data for each of the continuous outcome measures were transformed using either log10 (ocular sun exposure, HEL expression in tears and cells, and 8OHdG expression in tears) or square root (UVAF) transformations to approximate normal distributions. Several univariate relationships were identified; multivariate linear regression analysis supported significant relationships for each outcome variable: ocular sun exposure with outdoor occupation; tear HEL expression with increased tear 8OHdG expression and decreased cell HEL expression; cell HEL expression with decreased tear HEL expression, decreased contact lens wear, and inversely with age; tear 8OHdG expression with increased tear HEL expression, and inversely with age; total UVAF with decreased tear 8OHdG expression. Conclusions: We demonstrate feasibility and repeatability over a two week time period for ocular sun exposure and UVAF, and for previously unreported ELISA measurements of oxidative stress levels in human tears (8OHdG) and conjunctival cells (HEL). Our HEL ELISA findings for tears are notably higher than previous reports on human tears, possibly due to experimental differences. Results suggest that ocular surface lipid peroxidation is accompanied by DNA damage, lipid peroxidation of tears and conjunctival cells fluctuate inversely, and cellular changes causing UVAF are inhibitory toward DNA damage evidence in tears. Other factors that may influence expression of iii ocular surface oxidative stress biomarkers are gender, season of collection, contact lens wear, and UV exposure. iv Dedication This document is dedicated to my son, Ethan, for his inspiring unquenchable thirst for knowledge and insightful wisdom. v Acknowledgments I would like to thank my advisor, Heather Chandler, for her diligent assistance to coach me through laboratory analysis protocols, presentation development, and ultimately with this dissertation. Another tireless supporter, Karla Zadnik, has provided support for my research from her heart and by providing financial resources, and to her I will always be grateful. Thanks are also well deserved by Don Mutti, chief statistics consultant for my dissertation, whose enthusiasm for research is unrivaled. I am also thankful for the many wonderful friends I have made during my time as a graduate student who have provided me with untold levels of intellectual stimulation and emotional support. In summary, I will forever be grateful for each person I have had the privilege of knowing, that without my experience as a graduate student at The Ohio State University College of Optometry, would have left an incalculable void in my life. vi Vita 1964................................................................Born, Glendale, CA 1982................................................................High School Diploma, Fresno Adventist Academy 1986................................................................BBA Business Administration, Loma Linda University 1993................................................................OD Doctor of Optometry, Southern California College of Optometry 1993 to 2001 .................................................Clinical Optometrist, Fresno, CA 2001 to 2007 .................................................Optometric Practice Owner, Kristina M. Haworth, OD, Fresno, CA 2009...............................................................MS Vision Science, The Ohio State University 2007 to 2008 .................................................Graduate Teaching Associate, College of Optometry, The Ohio State University 2008 to 2011 .................................................Postdoctoral Fellow, College of Optometry, The Ohio State University vii 2011 to present ..............................................Assistant Professor - Practice, College of Optometry, The Ohio State University Publications Haworth KM and Chandler HL. Oxidative Stress Measures and Correlations with Lipid and DNA Damage Assays for Human Tears and Conjunctival Cells. (In preparation) Pucker AD and Haworth KM. The Presence & Significance of Polar Meibum & Tear Lipids. The Ocular Surface, accepted manuscript available online: 8-OCT-2014; DOI information: 10.1016/j.jtos.2014.06.002. Haworth KM, Nichols JJ, Thangavelu M, Sinnott LT, Nichols KK. Examination of human meibum collection and extraction techniques. Optometry and Vision Science Apr 2011;88(4):525-533. Peer-reviewed Abstracts Haworth KM and Chandler HL. Conjunctival Ultraviolet Autofluorescence and Ocular Sun Exposure. 2014. Accepted for presentation at the American Academy of Optometry annual meeting, Denver, CO, on Wednesday, November 12, 2014. Paper. viii Haworth KM and Chandler HL. Oxidative Stress Measures and Correlations with Lipid and DNA Damage Assays for Human Tears and Conjunctival Cells. 2014. Association for Research in Vision and Ophthalmology annual meeting, Orlando, FL. ARVO Meeting Abstracts April 30, 2014 55:2769. Poster. Mutti DO, Shiley KA, Haworth KM, Zadnik K. Dietary Vitamin D Consumption in Young Adult Myopes. 2011. Association for Research in Vision and Ophthalmology annual meeting, Fort Lauderdale, FL. ARVO meeting abstracts April 22, 2011 52:2702. Poster. Haworth KM, Chen J, Green-Church KB, Nichols KK. Comparison of Various Collection Techniques for Human Meibum with Mass Spectrometric Analysis. 2009. American Academy of Optometry annual meeting, Orlando, FL. American Academy of Optometry Archives #90725. Paper. Haworth KM, Nichols JJ, Thangavelu M, Sinnott LT, Nichols KK. Examination of Human Meibum Collection and Extraction Techniques. 2009. Association for Research in Vision and Ophthalmology annual meeting, Fort Lauderdale, FL. ARVO meeting abstracts April 11, 2009 50:2544. Paper. ix Fields of Study Major Field: Vision Science x Table of Contents Abstract ............................................................................................................................... ii Dedication ........................................................................................................................... v Acknowledgments.............................................................................................................. vi Vita .................................................................................................................................... vii Table of Contents ............................................................................................................... xi List of Tables ................................................................................................................... xvi List of Figures ................................................................................................................ xviii Chapter 1: Introduction ...................................................................................................... 1 1.1 Public Health Relevance and Project Significance .............................................. 1 Chapter 2: Background ....................................................................................................... 5 2.1 Ultraviolet Radiation Exposure ............................................................................ 5 2.1.1 Ultraviolet
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