Re-endothelization of human saphenous vein scaffold surfaces for bioprosthesis fabrication

Abstract

Various in vitro methods have been used for biological and synthetic scaffold fabrication. Some use polymers such as expanded polytetrafluoroethylene (ePTFE), polytetrafluoroethylene (PTFE), and polyethylene terephthalate (PET), while others use allogeneic or xenogeneic biological materials (e.g. blood vessels). While fabricating a biological scaffold, the first step is complete decellularization by enzymes (e.g. trypsin, collagenase, etc.) or chemicals (e.g. SDS, Triton-X, etc.), and the scaffolds should maintain its extracellular matrix (ECM). The second step involves re-endothelization so as to get fully biomimetic graft. In this study, we focused (concentrated) on the fabrication of a human saphenous vein scaffold by using various chemicals. We observed that cationic 1% SDS solution (Group B) performed excellent decellularization without altering the extracellular matrix as compared to the other chemicals like 0.25% trypsin and 70% ethanol (Groups C and D). Decellularization percentage and intactness of ECM (in all tunicae – intima, media, and adventitia) were confirmed based on histology. The PicoGreen assay showed that Group B (1% SDS decellularized scaffold, n = 3) had no detectable residual DNA. Re-endothelization on the complete decellularized scaffold (Group B) was done in both ways, without initial fibrin glue application (Group E) and with prior fibrin glue application (Group F). The vWF and lectin expressions suggested that endothelial cells did not alter their phenotype on human saphenous vein scaffolds. Uniaxial tensile testing revealed no significant differences in strain characteristics and modulus between native tissue and decellularized scaffolds. The live-dead (FDA/PI) and MTT assays confirmed the endothelial cell proliferation and viability, and the scanning electron microscope (SEM) data showed that the cells adhered to the scaffold matrix (Group F). We concluded that an allogeneic human saphenous vein scaffold with desirable properties can be fabricated and re-endothelialized to form a non-thrombogenic intimal surface in vitro using this protocol.