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J Thorac Cardiovasc Surg 1994;108:1043-1048
© 1994 Mosby, Inc.

SURGERY FOR ACQUIRED HEART DISEASE

Rapid visualization of viable and nonviable endothelium on cardiovascular prosthetic surfaces by means of fluorescent dyes

Stockholm, Sweden

Supported by the Swedish Heart & Lung Foundation, the Swedish Society for Medical Research, and the Swedish Society of Medicine, Konung Gustaf V:s and Drottning Victorias Stiftelse and the Swedish MRC (7126).

Received for publication Nov. 8, 1993. Accepted for publication June 2, 1994. Address for reprints: Caroline Gillis, MD, Karolinska Institute, Department of Neuroscience, Doktorsringen 17, S-171 77 Stockholm, Sweden.

Abstract

Increasing interest in endothelialization of synthetic and tissue cardiovascular prostheses in vitro emphasizes the need for simple and rapid methods to evaluate presence of endothelium on surfaces. Scanning electron microscopy is a commonly used method for this purpose. In this study we investigated alternative and more rapid staining methods. Human saphenous vein endothelial cells in culture and on cardiovascular prosthetic materials (pyrolytic carbon, cusps of bioprosthetic heart valves, pig aorta, and expanded polytetrafluoroethylene) were labeled by exposing them to medium containing 5-chloromethylfluorescein diacetate or 1,1-dioctadecyl-3,3,3',3'-tetramethylindo carbocyanine perchlorate. For comparison, specimens were also fixed and processed for scanning electron microscopy. A bright fluorescence of endothelial cells labeled with 5-chlormethylfluorescein diacetate or 1,1-deoctadecyl-3,3,3',3'-tetramethylindo carbocyanine perchlorate were clearly visualized in culture, on pyrolytic carbon, and on expanded polytetrafluoroethylene. Unfixed, prelabeled cells could be visualized immediately and unlabeled cells could be investigated for viability within 1 hour. Cells seeded on biologic tissue specimens could be visualized within 15 minutes with a modified hematoxylin-eosin staining. We suggest the use of these methods for rapid visualization of endothelium present on surfaces of cardiovascular prosthetic materials where they can partly replace the use of scanning electron microscopy. (J THORACCARDIOVASCSURG1994;108:1043-8)

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				C. Gillis, L. Bengtsson, and A. Haegerstrand REDUCTION OF MONOCYTE ADHESION TO XENOGENEIC TISSUE BY ENDOTHILIAZATION: AN ADHESION MOLECULE AND TIME-DEPENDENT MECHANISM J. Thorac. Cardiovasc. Surg., December 1, 1995; 110(6): 1583 - 1589. [Abstract] [Full Text]