Angiogenesis and Myogenesis in a Chronic Ischemic Heart. Esha Ibrahim East Tennessee State University
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East Tennessee State University Digital Commons @ East Tennessee State University Electronic Theses and Dissertations Student Works 8-2005 Angiogenesis and Myogenesis in a Chronic Ischemic Heart. Esha Ibrahim East Tennessee State University Follow this and additional works at: https://dc.etsu.edu/etd Part of the Medical Sciences Commons Recommended Citation Ibrahim, Esha, "Angiogenesis and Myogenesis in a Chronic Ischemic Heart." (2005). Electronic Theses and Dissertations. Paper 1058. https://dc.etsu.edu/etd/1058 This Thesis - Open Access is brought to you for free and open access by the Student Works at Digital Commons @ East Tennessee State University. It has been accepted for inclusion in Electronic Theses and Dissertations by an authorized administrator of Digital Commons @ East Tennessee State University. For more information, please contact [email protected]. Angiogenesis and Myogenesis in a Chronic Ischemic Heart A thesis presented to the faculty of the Department of Physiology East Tennessee State University In partial fulfillment of the requirements for the degree Master of Science in Biomedical Science by Esha Ibrahim August 2005 Race L. Kao, Ph.D., Chair Brian Rowe, Ph. D. Krishna Singh, Ph. D. Keywords: TMR, angiogenesis, myogenesis, myocardial ischemia, ventricular function ABSTRACT Angiogenesis and Myogenesis in a Chronic Ischemic Heart by Esha Ibrahim Miniswine underwent procedures to evaluate treating chronic ischemia with the implantation of autologous satellite cells and laser transmyocardial revascularization (TMR). The objective was to combine two therapies to restore cardiac function. This experiment involved three surgical procedures: (1) placing a constrictor on the coronary artery; (2) producing channels and implanting cells; (3) obtaining samples. The swine were divided into groups: Group 1, Ischemia; Group 2, Ischemia + Laser TMR; Group 3, Ischemia + Laser TMR+ Cells; Group 4, Ischemia + Cells. Sonomicrometry and Millar pressure transducers were used to determine contractility, left ventricular pressure, and pressure-volume loops. There were no significant differences (p<0.05) among the hemodynamic data except for Group 4, which produced significantly lower output values. Morphological evaluation revealed a significantly reduced scar area in Group 3. Although there was a significant difference in scar area, the phenomena behind this improvement as compared to the unimproved hemodynamic function is not understood. 2 CONTENTS Page ABSTRACT ..................................................................................................................... 2 LIST OF TABLES ........................................................................................................... 6 LIST OF FIGURES ......................................................................................................... 7 Chapter 1. INTRODUCTION .................................................................................................... 8 Transmyocardial Revascularization ...................................................................... 9 Angiogenesis Hypothesis ................................................................................ 9 Cellular Cardiomyoplasty ..................................................................................... 12 Satellite Cells: Identification and Significance ...................................................... 12 Satellite Cells: Muscle Repair................................................................................ 13 Rationale for Thesis .............................................................................................. 17 2. MATERIALS AND METHODS .............................................................................. 18 Materials .............................................................................................................. 18 Animals .......................................................................................................... 18 Satellite Cells .................................................................................................. 18 CO2 Heart Laser ............................................................................................ 18 Ameroid Constrictor ...................................................................................... 18 Gortex Surgical Membrane ............................................................................. 19 Media ............................................................................................................. 19 Enzymes ........................................................................................................ 19 Microspheres ................................................................................................. 19 3 Syringe Pump ................................................................................................. 19 Computer program hardware and software ..................................................... 20 Methods .............................................................................................................. 20 Study Design ....................................................................................................... 20 Experiment Time Line............................................................................................ 21 Animals .......................................................................................................... 22 Autologous Stem Cell Isolation, Cultivation, and Labeling ............................. 23 Myocardial Infarction ..................................................................................... 24 Cell Preparation for Implantation .................................................................... 24 Laser Transmyocardial Revascularization ....................................................... 25 Hemodynamic Evaluation ............................................................................... 26 Global Contractility ........................................................................................ 27 Cardiac Output ........................................................................................ 28 Ejection Fraction ...................................................................................... 28 Regional Contractility ..................................................................................... 28 Regional Perfusion ......................................................................................... 29 Statistical Analysis ......................................................................................... 30 Heart Harvest ................................................................................................. 31 Heart Sectioning ............................................................................................ 31 3. RESULTS ................................................................................................................ 32 Isolation of Satellite Cells from the Skeletal Muscle of Miniswine ........................ 32 Cell Counts .................................................................................................... 32 Pathological Evaluations ...................................................................................... 33 Electrocardiograms .............................................................................................. 35 Hemodynamic Evaluations ................................................................................... 37 4 Cardiac Output ............................................................................................... 37 Ejection Fraction ............................................................................................ 38 Segment Shortening ....................................................................................... 39 Pressure-Volume Loops ................................................................................. 40 Regional Perfusion ......................................................................................... 47 4. DISCUSSION ......................................................................................................... 48 Limitations ........................................................................................................... 52 Conclusion ........................................................................................................... 53 REFERENCES ............................................................................................................ 54 VITA ………............................................................................................................... 58 5 LIST OF TABLES Table Page 1. Stroke Volume Values and Averages per Animal and per Animal Group ................. 42 2. Load Variables and Averages per Animal and per Animal Group ............................ 46 3. Microsphere Counts to Measure Regional Perfusion ............................................... 47 6 LIST OF FIGURES Figure Page 1. Laser Handpiece in Contact with Myocardium ......................................................... 9 2. Proposed Mechanism for TMR Effectiveness ........................................................... 10 3. Satellite Cell Location .............................................................................................. 14 4. Sternatomy Procedure Showing Labeling of Crystals, Constrictor, and Transducers .. 26 5. Pressure-Volume Loop Diagram .............................................................................