THE REFERENCE MATERIALS OF THE EUROPEAN COMMUNITIES The Reference Materials ofthe European Cornrnunities Results of Hemocompatibility Tests edited by W.Lemm EUROBIOMAT Research Program, Berlin, Germany SPRTNGER-SClENCE+BUSINESS MEDIA, B.V. Library of Congress Cataioging-in-Publication Data The Reference materlals of the European communltles results of hemocow-pat 1t l il; ty Tests ed 1t or W. Lelllm. p. cm. ISBN 978-90-481-4213-2 ISBN 978-94-015-7993-3 (eBook) DOI 10.1007/978-94-015-7993-3 1. Blocolllpat1tli 11t~. 2. Blood--Effect of ll1plants on. 1. Lelllm. W. R857.M3R44 1992 610' .28--dc20 92-30458 ISBN 978-90-481-4213-2 Printed on acid-jree paper AII Rights Reserved © 1992 Springer Science+Business Media Dordrecht Originally published by K1uwer Academic Publishers in 1992 No part of the material protected by this copyright notice may be reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording or by any information storage and retrieval system, without written permission from the copyright owner. FOREWORD The present book continues the number of pUblications which has been released by the European Concerted Action "EUROBIOMAT - Hemocompatibility" (Medical Research Program, Project: 11.1.2/2). This Concerted Action has been supported from 1988-1992 by the European communities in order to promote the collaboration between the majority of European countries on science and technology on the particular field of hemocompatible biomaterials. The most serious problem in developing better hemocompa tible biomaterials is the absence of standardised in-vitro, ex-vivo, and in-vivo test methods and the deficiency of at least generally accepted test procedures for biomaterials and their biocompatibility to compare the test results between the different laboratories. Therefore Reference Materials were distributed to several research centers in order to perform a variety of test procedures on identical surfaces. This book is the outcome of a workshop held in Copenhagen from 31 January - 2 February 1992. 1t presents the results of tests performed on the surfaces of the Reference Materials of the European Communities. This book is subdivided in two chapters: - Surface and bulk characterisation by physical, physico-chemical and chemical methods. - Biological tests such as toxicity tests and hemocompatibility tests (in-vitro, ex-vivo and in-vivo) . v Table of Contents Foreword v The Concerted Action EUROBIOMAT -Hemocompatibility- xi Introduction xv General Information and Instructions xxi Part I physical, Chemical and Physico-chemical Methods for Surface and Bulk Characterisation Surface Characterization of EUROBIOMAT Reference Polymers. castner, D.G., Ratner, B.D., Leach-Scampavia, D.K. 3 Surface Analysis of Polymer Biomaterials by Electron Spectroscopy for Chemical Analysis (ESCA). Bauser, H., Gunsilius, H., Hellwig, G. 17 Surface Infrared Analysis of Standard Biomaterials. Barbucci, R., Magnani, A. 27 Contact Angles on Polymer Reference Materials. Costa, M., Ringard, C., Boissonnade, M.-M., Baszkin, A. 37 Zeta-Potentials and Surface Tensions. Mientus, w., Zecha, Ch., Olschewski, M. 49 The Surface Energy Spectra of Polymers. Mash, J. 65 viii Supercritical Fluid Extraction / Chromatography (SFE/C) of Additives from Standard Biomaterials for Use in Biocompatibility Testing. Braybrook, J.H., Mackay, G.A. 75 Determination of Surfaee Energetic Properties by Contact Angle Measurements. Glasmacher-Seiler, B., Voigt, S., Reul, H. 85 Advancing and Reeeding Water Contaet Angle Measurements with Drops, Bubbles and Plates. Kamusewitz, H., Possart, W., Paul, D. 95 Screening Test for Residual Ethylene Oxide on Reference Materials. Dawids, G.S., Christoffersen, F., Elhauge, T. 107 The Average Molecular Weight of the Referenee Materials. Lemm, W. 113 Part 11 Biologieal, Biochemieal, Immunologieal, Hematologieal and Toxieologieal Methods to Evaluate the Bioeompatibility of the Referenee Materials Activation of the Human Complement System by "Standard" Polymers. Maillet, F., Kazatchkine, M.D. 121 Photometrie Enzyme Immunoassay for the Quantitative Determination of C3a-des Arg in Human Plasma Ineubated with Polymerie Biomaterials. Krämer, S., Burger, R., Weiss, P., Malseh, G. 127 complement Aetivation, Cytotoxieity and Leaehing of Substanees. Brown, G.S., Rychly, J., Thomaneck, U., Klinkmann, H. 139 ix Enzyme-Linked Immunosorbent Assays for Biocompatibility Testing. Hall, B., Pearce, D.J., Campbell, E.J., Sullivan, A.M., New, R.R.C., Charles, S.A. 147 Measurement of Platelet Adhesion, released ß-Thromboglobulin and Generated Fibrinopeptide A Using whole Non-anti-coagulated Blood at Flow Conditions. Missirlis, Y., Michanetzis, G. 157 Perfusion Technique for in vitro Evaluation of Platelet Interaction with Biomaterials in Catheter Form. Mulvihill, J.N., Cazenave J.-P. 165 In-vitro Hemocompatibility Screening Test by Using the Resonance-Thrombography. Lemm, w. 173 The Hemocompatibility of Reference Materials in-vitro; Comparative Investigation of Platelet AdhesionjActivation and Platelet Aggregability. Groth, Th., Gronert, A., Ziemer, S., Hesse, R. 183 Cell Culture Cytotoxicity Assays as Part of the Biocompatibility Testing of Standard Biomaterials. Braybrook, J.H., Doherty, P. 193 Quantitative Evaluation of Cytotoxicity in vitro of Extracts from Standard Biomaterials. Dekker, A., Hermanns, M. I • , Valdor, M. , Kirkpatrick, C.J. 201 In-vitro Toxicity of the Reference Materials. The Ciliata-Test. Lemm, W. 211 Keratinocyte Cell Cultures for Testing Polymers for Percutaneous Devices. Nühlen, u., Große-Siestrup, C. 217 x Blood-Testing of a Polyether-Urethane as a Diffusion Limiting and Potentially Haemocompatible External Membrane for Enzyme Electrode Biosensors. Crump, P.W., Ghosh, S., Vadgama, P. 227 Biocompatibility Evaluation of Reference Materials. Eloy, R., Charton Picard, F., Belleville, J., Bouet, T. 237 The Concerted Action EUROBIOMAT- Hemocompatibility. The Concerted Action -EUROBIOMAT- was initiated in early 1988 by the Commission of the European Communities and is one among 28 other Concerted Actions covering the scientific and technological aspects of biomedical engineering. EUROBIOMAT is precisely entitled "Replacement of Body Parts and Functions" and is focused on the problems related to "Hemocompatibility". Approximately 50 European centers are gathered in this cooperative project. The demand for hemocompatibility is one of the basic factors for a safe and sufficient application of artificial surfaces being in contact with blood either permanently or temporarily. Thrombus formation and infarctions of the capillary vascular system after blood material interaction as weIl as the activation of the complement system and the phenomenon of biodegradation are still unsolved problems. However, medical devices where large areas of such artificial surfaces contact the blood of patients are applied in enormous quantities. In spite of the high significance of this branch of modern medicine and certainly spectacular success in therapy, organ support, and organ replacement, clearly specified criteria, guidelines and instructions for the design of devices and artif icial blood contacting surfaces and their safe application do not yet exist. Standardised and generally accepted test procedures in order to quantify and validate precisely the events on the blood-material- interface were up to now not available. These circumstances limited a safe medical therapy as weIl as a more efficient development of new and better hemocompatible blood contacting polymeric surfaces. The missing success in the development of new and better hemocompatible biomaterials is due to the absence of generally accepted and standardised test procedures to qualify blood contacting surfaces. xi xii within the frame -Hemocompatibility- the participants started with the collection of precisely described test-procedures in order to standardise them. The outcome of this sub-project is the edition of the book of standards covering all aspects of biomaterials testing. It is entitled: Test Procedures for the Blood Compatibility of Biomaterials. A cooperation with the International Standards Organisation (ISO) within the project TC 194 (Biological Evaluation of Medical and Dental Materials and Devices) started in order to elaborate standards for biomedical engineering in particular on the field of biodegradation, hemocompatibility tests and sample preparation and reference materials. In the following step a Network of European Test Centers has been established where approximately 25 centers are actively involved each of them contributing a particular fragment of biomaterials testing in order to compare test results, to evaluate the limitations of individual test procedures and to qualify and certify biocompatible materials and surfaces. In the next step six different Reference Materials were purchasedi they are available either as tubes or films. These materials are already widely in medical use. Each of these six materials was produced under identical conditions in medical-grade quality from one single batch in larger quantities in order to provide the Network of European Test Centers and any new interested research center with the standard surfaces to control their test facilities. This book presents the results of the tests performed on the identical surfaces of the Reference Materials of the European Communities. After this pioneer work several research centers started wi th the development of new and better hemo- and bio compatible biomaterials or material modifications.