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      Thrombomodulin Activity of Fat-Derived Microvascular Endothelial Cells Seeded on Expanded Polytetrafluorethylene

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          Abstract

          Lining the luminal surface of prosthetic vascular grafts with endothelial cells (cell seeding) will lower its thrombogenicity. Commonly used macrovascular human adult endothelial cells (HAEC) require in vitro cultivation before large enough numbers are obtained to cover grafts confluently. Fat-derived microvascular endothelial cells (MVEC) prove to be a good alternative as they can be harvested in much larger numbers while showing similar antithrombotic and fibrinolytic characteristics. An important anticoagulant function of macrovascular endothelial cells is due to the activity of thrombomodulin (TM) on their surface. In this study, the presence and functional activity of TM on fat-derived microvascular cells used in cell seeding was investigated. The expression and localization of TM on MVEC was studied using immunohistochemistry. Functional activity of TM on MVEC was measured by the generation of activated protein C (APC) and was compared to human umbilical vein endothelial cells (HUVEC). TM activity was studied in MVEC seeded on expanded polytetrafluorethylene (ePTFE) vascular prostheses and compared to blank prostheses. We found that TM was expressed on the surface of MVEC, both in vivo and vitro. TM-dependent generation of APC differed significantly between MVEC and HUVEC (3.98 ± 1.2 vs. 3.0 ± 0.7 n M, respectively). After seeding MVEC on vascular prostheses, TM activity did not change. APC generation was significantly higher on MVEC-seeded vascular grafts compared to blank grafts (4.0 ± 0.7 vs. 1.7 ± 0.5 n M, respectively). We conclude that TM is present and highly active on cultured MVEC. When seeded on ePTFE, MVEC retain the possibility to inhibit thrombin coagulant activity and to activate protein C. Therefore, since MVEC are readily available, the anticoagulant properties demonstrated here indicate that this cell type is suitable for cell seeding of vascular prostheses.

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          Six-year prospective multicenter randomized comparison of autologous saphenous vein and expanded polytetrafluoroethylene grafts in infrainguinal arterial reconstructions

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            Endothelial seeding of polytetrafluoroethylene popliteal bypasses. A preliminary report.

            At St. Vincent Hospital, Indianapolis, 17 patients have undergone femoropopliteal bypass operations with polytetrafluoroethylene (PTFE) grafts that were seeded with enzymatically harvested, autogenous endothelium. Three patients received seeded grafts because satisfactory veins were not available. Twenty-eight patients alternately received seeded or unseeded, externally supported e-PTFE grafts. Graft patency was evaluated by clinical criteria and changes in the Doppler ankle-brachial systolic pressure ratios at 2 and 30 days postoperatively and at 3-month intervals thereafter. Occlusions were defined arteriographically if the clinical situation or the Doppler findings deteriorated. Smoking histories were taken, and carboxyhemoglobin (COHgb) levels were sampled 1 month postoperatively. Cumulative patency after 3 months was 93.3% +/- 6.5% for seeded and 84.0% +/- 10.4% for unseeded grafts. After 1 year it was 81.6% +/- 12.3% for seeded grafts and 30.8% +/- 18.7% for unseeded grafts (p = 0.02). Thus far all but one of the occlusions have occurred in patients with a history of smoking or with a COHgb level greater than 1.5%, whereas all of the seeded grafts in nonsmokers with COHgb levels less than or equal to 1.5% are patent. We conclude that endothelial seeding of PTFE femoropopliteal grafts is feasible. During this preliminary study period, a small number of patients had favorable patency rates with seeded as compared with unseeded grafts, especially among smokers. A multiple-institution study will be needed to establish the role of endothelial cell seeding in the treatment of vascular occlusions of the femoral and popliteal arteries.
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              Author and article information

              Journal
              JVR
              J Vasc Res
              10.1159/issn.1018-1172
              Journal of Vascular Research
              S. Karger AG
              1018-1172
              1423-0135
              1999
              April 1999
              22 April 1999
              : 36
              : 2
              : 91-99
              Affiliations
              Departments of Surgery and Hematology, University Hospital Utrecht, The Netherlands
              Article
              25630 J Vasc Res 1999;36:91–99
              10.1159/000025630
              10213903
              © 1999 S. Karger AG, Basel

              Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher. Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug. Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

              Page count
              Figures: 6, References: 33, Pages: 9
              Categories
              Research Paper

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