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J Thorac Cardiovasc Surg 2003;126:455-464
© 2003 The American Association for Thoracic Surgery

Cardiopulmonary support and physiology

Canine endothelial progenitor cell-lined hybrid vascular graft with nonthrombogenic potential

^a Department of Biomedical Engineering, Fukuoka, Japan
^b Department of Cardiac Surgery, Graduate School of Medicine, Kyushu University, Fukuoka, Japan
^c Department of Bioengineering, National Cardiovascular Center Research Institute, Osaka, Japan

Received for publication March 22, 2002; revisions received May 16, 2002; revisions received May 29, 2002; accepted for publication July 26, 2002.

^* Address for reprints: Takehisa Matsuda, MD, Department of Biomedical Engineering, Graduate School of Medicine, Kyushu University, 3-1-1 Maidashi, Higashi-ku grbf, Fukuoka 812-8582, Japan
matsuda{at}med.kyushu-u.ac.jp

OBJECTIVE: We sought to fabricate a compliant engineered vascular graft (inner diameter of approximately 4.5 mm and length of 6 cm) lined with endothelial progenitor cells derived from circulating peripheral canine blood and to verify its nonthrombogenicity potential in vivo.

METHODS: Autologous circulating endothelial progenitor cells derived from the peripheral veins of 6 adult mongrel dogs were isolated by using a density gradient method. The cells were proliferated in vitro in EGM-2 culture medium, prelined on the luminal surface of in situ–formed collagen type I meshes as an extracellular matrix, and wrapped with a segmented polyurethane thin film with multiple micropores as a compliant scaffold. After canine carotid arteries were bilaterally implanted with these grafts for 1 and 3 months, microscopic observation, histologic staining, and immunochemical staining were performed to evaluate morphogenesis.

RESULTS: After 33.3 ± 10.5 days of culture in vitro, 4.2 ± 1.2 x 10⁶ endothelial progenitor cells were obtained. Eleven of the 12 engineered vascular grafts were patent. The grafts possessed smooth, glistening, and ivory-colored luminal surfaces at the predetermined observation period up to 3 months. The intimal layer was covered with confluent, cobblestone-like monolayered cells that were positively stained with factor VIIIB–related antigen. The thickness of the neoarterial walls was approximately 300 µm at 3 months after implantation. A few smooth muscle cells were observed in the medial tissue, and fibroblasts dominated the adventitial tissue.

CONCLUSION: Circulating endothelial progenitor cells could be a substitute source of endothelial cells for endothelialization on small-diameter-vessel prostheses to ensure nonthrombogenicity.