The decellularized porcine heart valve matrix in tissue engineering: platelet adhesion and activation.
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-I (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.
Kasimir
Marie-Theres
MT
Department of Cardiothoracic Surgery, Medical University of Vienna, Vienna, Austria.
Weigel
Guenter
G
Sharma
Jyotindra
J
Rieder
Erwin
E
Seebacher
Gernot
G
Wolner
Ernst
E
Simon
Paul
P
D000818:Animals; D001021:Aortic Valve / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D001705:Bioprosthesis / Q000009:adverse effects; D001792:Blood Platelets / Q000378:metabolism*; D002478:Cells, Cultured; D018920:Coculture Techniques; D042783:Endothelial Cells / Q000378:metabolism / Q000637:transplantation*; D005109:Extracellular Matrix / Q000378:metabolism*; D006349:Heart Valve Diseases / Q000628:therapy; D006350:Heart Valve Prosthesis / Q000009:adverse effects*; D006801:Humans; D015539:Platelet Activation; D010973:Platelet Adhesiveness; D013501:Surface-Active Agents / Q000494:pharmacology; D013552:Swine; D013927:Thrombosis / Q000209:etiology / Q000517:prevention & control; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods*
D013501:Surface-Active Agents