Thrombogenicity, which is commonly encountered in artificial heart valves after replacement surgeries, causes valvular failure. Even life-long anticoagulant drug use may not be sufficient to prevent thrombogenicity. In this study, it was aimed to develop a heart valve construct with antithrombogenic properties and suitable mechanical strength by combining multiwalled carbon nanotubes within a decellularized bovine pericardium. In this context, the decellularization process was performed by using the combination of freeze–thawing and sodium dodecyl sulfate (SDS). Evaluation of decellularization efficiency was determined by histology (Hematoxylin and Eosin, DAPI and Masson’s Trichrome) and biochemical (DNA, sGAG and collagen) analyses. After the decellularization process of the bovine pericardium, composite pericardial tissues were prepared by incorporating −COOH-modified multiwalled carbon nanotubes (MWCNTs). Characterization of MWCNT incorporation was performed by ATR-FTIR, TGA, and mechanical analysis, while SEM and AFM were used for morphological evaluations. Thrombogenicity assessments were studied by platelet adhesion test, Calcein-AM staining, kinetic blood clotting, hemolysis, and cytotoxicity analyses. As a result of this study, the composite pericardial material revealed improved mechanical and thermal stability and hemocompatibility in comparison to decellularized pericardium, without toxicity. Approximately 100% success is achieved in preventing platelet adhesion. In addition, kinetic blood-coagulation analysis demonstrated a low rate and slow coagulation kinetics, while the hemolysis index was below the permissible limit for biomaterials.
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