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J Thorac Cardiovasc Surg 1994;108:540-548
© 1994 Mosby, Inc.

SURGERY FOR ACQUIRED HEART DISEASE

Release of platelet-derived growth factor activity from pig venous arterial grafts

Sheffield and Cardiff, United Kingdom

Supported by grants from the National Heart Research Fund and the British Heart Foundation.

Received for publication Oct. 13, 1993. Accepted for publication Jan. 28, 1994. Address for reprints: Department of Cardiac Surgery, University of Bristol, Bristol Royal Infirmary, Bristol BS2 8HW, United Kingdom.

Abstract

Intimal smooth muscle cell proliferation and superimposed atheroma are the main causes of late failure of saphenous vein bypass grafts. It has been suggested that these reactions are caused by the production of growth factors from the cells of the vessel wall. To test this hypothesis, we cultured segments of pig venous arterial grafts, removed 1 and 4 weeks after implantation, in serum-free medium for 24 hours. Tissue viability as assessed by adenosine triphosphate concentration was maintained throughout the 24-hour culture period (239±21 nmol/gm wet weight [standard error of the mean], n = 26, 0 hours; 240±24 nmol/gm wet weight, n = 17, 24 hours). Cell proliferation occurred and autoradiography showed proliferating cells to be located in the neointimal and medial layers. These cells were identified as smooth muscle cells by means of a monoclonal antibody to-actin. Graft-conditioned media were tested for mitogenic activity by means of a fibroblast proliferation assay. Media conditioned for 24 hours produced significant stimulation of cell growth (284%±30%, n = 17) above that obtained in culture medium alone (100%). This mitogenic activity was inhibited by 61%±9%, n = 8, with a polyclonal-neutralizing antibody to platelet-derived growth factor. Reverse-transcription polymerase chain reaction analysis and Northern blots demonstrated platelet-derived growth factor B messenger ribonucleic acid (mRNA) in vein grafts but not in ungrafted vein. Analysis of graft tissue sections by in situ hybridization demonstrated an abundance of platelet-derived growth factor B mRNA positive cells in the endothelial and neointimal layers, as well as in the endothelial cells of the adventitial vessels. These data constitute direct evidence for active growth factor production within the cells of the vein graft. They also suggest that endogenously produced platelet-derived growth factor may play a role in regulating smooth muscle cell proliferation in this model. (J THORACCARDIOVASCSURG1994;108:540-8)

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