The decellularized porcine heart valve matrix in tissue engineering

Marie-Theres Kasimir; Guenter Weigel; Jyotindra Sharma; Erwin Rieder; Gernot Seebacher; Ernst Wolner; Paul Simon

doi:10.1160/TH05-01-0025

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2005; 94(03): 562-567
DOI: 10.1160/TH05-01-0025

Platelets and Blood Cells

Schattauer GmbH

The decellularized porcine heart valve matrix in tissue engineering

Platelet adhesion and activation

Marie-Theres Kasimir

¹Department of Cardiothoracic Surgery, Medical University of Vienna, Vienna, Austria

,

Guenter Weigel

¹Department of Cardiothoracic Surgery, Medical University of Vienna, Vienna, Austria

,

Jyotindra Sharma

¹Department of Cardiothoracic Surgery, Medical University of Vienna, Vienna, Austria

,

Erwin Rieder

²Department of Surgery, Medical University of Vienna, Vienna, Austria

,

Gernot Seebacher

¹Department of Cardiothoracic Surgery, Medical University of Vienna, Vienna, Austria

,

Ernst Wolner

¹Department of Cardiothoracic Surgery, Medical University of Vienna, Vienna, Austria

,

Paul Simon

¹Department of Cardiothoracic Surgery, Medical University of Vienna, Vienna, Austria

› Author Affiliations

Abstract

Summary

An approach in tissue engineering of heart valves is the use of decellularized xenogeneic matrices to avoid immune response after implantation. The decellularization process must preserve the structural components of the extracellular matrix to provide a biomechanically stable scaffold. However, it is known that in vascular lesions platelet adhesion to extracellular matrix components occurs and platelet activation is induced. In the present study we examined the effects of a decellularized porcine heart valve matrix on thrombocyte activation and the influence of re-endothelialisation in vitro. Porcine pulmonary conduits were decellularized using Triton X-100, Na-deoxycholate and Igepal CA-630® followed by a ribonuclease digestion. Cryostat sections of decellularized heart valves with and without seeding with human umbilical vein endothelial cells (HUVEC) were incubated with platelet rich plasma. Samples were either stained with fluorescent antibodies for CD41 and PAC-1 (recognizing the activated fibrinogen receptor) or fixed with glutaraldehyde. Thereafter, the samples were processed for laser scanning microscopy (LSM) or scanning electron microscopy (SEM). Examination by LSM showed numerous platelets with co-localized staining for CD41 and PAC-1 on the nonseeded decellularized heart valve matrix whereas after seeding with endothelial cells no platelet activation was detected. SEM revealed platelet adhesion and aggregate formation only on the surface of the non-seeded or partially denuded matrix specimens. We show in this study that the decellularized porcine matrix acts as a platelet-activating surface. Seeding with endothelial cells effectively abolishes the platelet adhesion and activation and therefore is necessary to eliminate thrombogenicity in tissue engineered heart valves.

Keywords

Biotechnology - tissue engineering - heart valves - decellularized - porcine matrix - thrombogenicity

Full Text

References

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