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J Thorac Cardiovasc Surg 2001;121:0108-0115
© 2001 The American Association for Thoracic Surgery

Surgery for Acquired Cardiovascular Disease

In vitro endothelialization of bioprosthetic heart valves provides a cell monolayer with proliferative capacities and resistance to pulsatile flow

From Karolinska Institute, Department of Surgical Sciences, Karolinska Hospital,^a Stockholm, Sweden; Hjertecenter,^b Varde, Esbjerg, Denmark; and Neuronova AB,^c Stockholm, Sweden.

This work was supported by grants from the Swedish Heart and Lung Foundation, the Swedish Medical Research Council (project No. 09100, 07126), Lars Hiertta Foundation, Lisa and Johan Grönbergs Foundation. The research on this topic was also supported by an unrestricted grant from TERUMO EUROPE N.V., dedicated to well-being.

Received for publication May 25, 2000. Revisions requested June 22, 2000; revisions received July 11, 2000. Accepted for publication July 18, 2000. Address for reprints: Katarina Jansson, BSc, Karolinska Institute, Department of Surgical Sciences, Research Center M3:01, Vascular Biology, Karolinska Hospital, S-171 76 Stockholm, Sweden (E-mail: katarina.jansson{at}kirurgi.ki.se).

Objectives: Degeneration of bioprosthetic heart valves has been suggested to be at least partly an immunogenic reaction toward the xenogeneic tissue. An autologous endothelial lining has been proposed to overcome this problem. We examined in vitro endothelialization of such tissue and retention of endothelial cells after exposure to flow resembling the in vivo situation.
Methods: Cultured human saphenous vein endothelial cells were used to in vitro endothelialize photo-oxidized bioprosthetic heart valves. The endothelialized valves were mounted in a specially designed flow device, creating a pulsatile flow through the valve. Maintenance of a confluent cell layer and deposition of basement membrane markers were determined with immunohistochemical labeling.
Results: Labeling of the main components of the basement membrane, laminin and collagen type IV, was verified within 6 hours after in vitro endothelialization. Under static conditions, 4-mm wide denudations were completely re-endothelialized in 4 days, which was similar to the growth rate on gelatin-coated cell culture plastic, which served as a control material. After exposure of endothelialized valves to pulsatile flows for 24 hours (80 beats/min, 3.4 L/min), there were minimal cell losses from the bioprostheses. The cell layer adapted to the pulsatile flow, as verified by rearrangement of morphology and intracellular stress fibers.
Conclusions: This study shows the feasibility of in vitro endothelialization of photo-oxidized bioprosthetic heart valves. The cells are able to withstand a pulsatile flow in vitro, to develop basement membrane–like structures, and to re-endothelialize denuded areas. This technology may be used to enhance the performance of bioprosthetic heart valve prostheses.