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INFORMATION TO USERS This manuscript has been reproduced from the microfilm master. UMI films the text directly from the original or copy submitted. Thus, some thesis and dissertation copies are in typewriter free, i ^ e others may be from any type of computer printer. The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleedthrough, substandard margins, and improper alignment can adversely afreet reproduction. In the unlikely event that the author did not send UMI a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion. Oversize materials (e.g., maps, drawings, charts) are reproduced by sectioning the original, beginning at the upper left-hand comer and continuing from left to right in equal sections with small overlaps. Each original is also photographed in one exposure and is included in reduced form at the back of the book. Photographs included in the original manuscript have been reproduced xerographically in this copy. Higher quality 6” x 9” black and white photographic prints are available for any photographs or illustrations appearing in this copy for an additional charge. Contact UMI directly to order. UMI A Bell & Howell Information Compaiy 300 North Zeeb Road, Ann Arbor MI 48106-1346 USA 313/761-4700 800/521-0600 PHYSIOLOGIC RESPONSES TO INFLAMMATION IN ISOLATED EQUINE JOINTS DISSERTATION Presented in Partial Fulfilment of the Requirements for the Degree of Doctor of Philosophy in the Graduate School of The Ohio State University B y Joanne Hardy, D.V.M., M.S. ÿ >{c ÿ ifc 3|c The Ohio State University 1996 Dissertation Committee: A.L. Bertone, Adviser W.W. Muir S.E. Weisbrode A dviser T.M. O’Dorisio Department of Veterinary Clinical Sciences D.N. Granger UMI Number: 9710576 UMI Microform 9710576 Copyright 1997, by UMI Company. All rights reserved. This microform edition is protected against unauthorized copying under Title 17, United States Code. UMI 300 North Zeeb Road Ann Arbor, MI 48103 ABSTRACT Articular inflammation is a common finding in equine joints disorders. As in other organs, several aspects of the joint system, including blood flow, transsynovial permeability and fluid exchanges, cellular infiltration and mediator release may be involved in the inflammatory process and contribute to joint morbidity. With inflammation, tissues within the joint contribute to mediator release and stimulate degradation of the cartilage matrix. Perfused oxygenated extracorporeal isolated models have been thoroughly described for organs such as heart, lung and intestine, but have never been created for the joint organ. The studies reported here describe an isolated pump-perfiised auto-oxygenated extracorporeal isolated joint model developed in the equine metacarpophalangeal joint. Using this model, the local Starling’s forces in the isolated joint at isogravimetric state are characterized and alterations in Starling’s forces in response to hemodynamic manipulations are described. In a separate series of experiments, articular inflammation was induced in the isolated innervated or denervated joint organ and local hemodynamic, fluid exchanges, metabolic responses, alterations in permeability to small and large molecules, inflammatory mediator release, cellular events and articular cartilage metabolic alterations are described. Two preliminary studies were performed in the development of the isolated joint model. The first studies was essential to gain a better understanding of the local biomechanical behavior of the fluid filled closed joint cavity. In this study, the investigation of pressure volume curves in the joint revealed a sigmoid relationship, with low compliance at normal subatmospheric pressures, and gradually increasing compliance at supraatmospheric pressures of up to 30 mmHg. Thereafter, compliance of the joint again decreased exponentially. This study also revealed an important function of synovial fluid to increase joint compliance and hysteresis therefore preventing collapse of the joint cavity and allow fluid exchanges at subatmospheric pressures. Furthermore this study emphasized the importance of joint angle on pressure-volume relationships. In the second preliminary study, a colored microsphere technique was validated for determination of blood flow to the synovial membrane and joint capsule. Furthermore, that study documented a significant decrease in synovial membrane blood flow down to 16% of baseline at intraarticular pressures of 30 mmHg or greater. The development and physiologic responses of the isolated model were subsequently described. The extracorporeal pump perfused isolated joint unit developed in this model was successfully be maintained for 6 hours. Using this system, isogravimetric state was defined at a circuit arterial pressure of 133 mmHg and venous pressure of 10.5 mm Hg. Starling’s forces were defined for this system. Arterial and venous pressure manipulations revealed that increases in arterial pressure significantly raised transsynovial flow and synovial fluid production, when lAP was maintained at atmospheric pressure. A similar but much less important response was observed with increased venous pressure. Acute inflammation was successfully induced in the isolated innervated or denervated joint preparation, using interleukin-IB. Acute inflammation induced a significant increase in oxygen consumption and extraction ratio in response to increased metabolic demand. Acute inflammation increased synovial membrane permeability to albumin, but denervation lowered permeability to large molecules (MW 144,(XX)). Inflammation significantly increased synovial fluid production and the permeability surface area product of the synovial membrane Acute inflammation was associated with a significant cellular response characterized by neutrophilic leukocytosis in synovial fluid and a neutrophilic vasculitis in the synovial membrane. A significant increase in interleukin-6, prostaglandin 5% and substance P was ui detected in synovial fluid collected from inflamed joints. Inflammation induced a significant degradative response in articular cartilage, even after this short term exposure to an inflammatory mediator. The isolated joint model described in these studies provided new insights in our understanding of articular physiology, an allowed to study of all pathophysiologic processes involved in articular inflammation. This model should be useful to further our knowledge of the response of joint to various disease processes, and should be very suitable to study the pharmacokinetics and pharmacodynamics of systemic or intraarticular therapies. IV ACKNOWLEDGMENTS I express sincere appreciation to Dr Alicia L Bertone for her guidance and enthousiasm throughout this research, and Dr William W Muir for his great insight, ideas and support Thanks go to Dr Steve Weisbrode, for his enthousiasm and for teaching me to understand histopathology. Thanks go to the other members of my advisory committee. Dr Tom O’Dorisio and Dr Neil Granger, for their time, suggestions and comments. The technical assistance of Mrs Kena Chapman, Mrs Robin Nakkula, and Ms Heather Burkhardt is gratefully acknowledged. VITA September 17th, 1957 .....................................Bom- Montreal, Quebec, Canada 1982 ............................................................... D.V.M., University of Montreal, Montreal, Quebec, Canada 1983 ................................................................. 1.P.S.A.V., Internat de Perfectionnement en Sciences Appliquées Vétérinaires, University of Montreal, Montreal, Quebec, Canada 1989 ...............................................................Residency, Equine Surgery, The Ohio State University, Columbus, Ohio 1991 Diplomate, American College of Veterinary Surgeons PUBLICATIONS Moore RM, Hance S, Hardy J. Colonic luminal pressure in horses with strangulating and nonstrangulating obstruction of the large colon. Vet Surg 1996; 25:134-141. Hardv J. Bertone AL, Muir WW. Joint pressure influences synovial tissue blood flow as determined by colored microspheres. J Appl Physiol 1996; 80:1225-1232. Beard WL, Hardy J. Techniques de fixation de fractures des incisives par cerclage interdentaire chez le cheval. Pratique Vétérinaire Equine 1995; 27:159-166. Moore RM, Hardy J. Muir WW. Mural blood flow distribution in the large colon of horses during low-flow ischemia and reperfusion. Am J Vet Res 1995,56: 812-818. Hardy J. Bertone AL, Muir WW. Pressure-volume relationships in the equine carpus. J Appl Physiol 1995, 78:1977-1984. Hardy J. Bednarski RM, Biller DS. Effect of phenylephrine on hemodynamics and splenic dimensions in the horse. Am J Vet Res 1994,55: 1570-1578. VI Carter BG, Schneider RK, Hardy J. Bramlage LR and Bertone AL. Assessment and treatment of equine humerai fractures: retrospective study of 54 cases (1972-1990). Equine VetJ 1993; 25; 3:203-207 Hardy J. Bertone AL. Surgery of the equine large and small colon. Compend Cont Educ Pract Vet 1992; 14(11): 1501-1506. Hardy J. Stewart RH, Beard WL, Yvorchuk K. Complications of nasogastric intubation in the horse: 9 cases (1987-1989). J Am Vet Med Assoc 1992; 201 (3); 483-486. Hardy J. Robertson JT, Reed SM. Pericardiectomy for treatment of constrictive pericarditis in a mare. Equine Vet J 1992; 24. Giraudet A, Hardy J. Reparation chirurgicale des déchirures du col utérin chez la jument. Technique et
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