Background: Poly (ethylene glycol) (PEG) has attracted broad interest for tissue\r\nengineering applications. The aim of this study was to synthesize 4-arm -PEG-20kDa\r\nwith the terminal group of diacrylate (4-arm-PEG-DA) and evaluate its dual\r\nfunctionality for decellularized porcine aortic valve (DAV) based on its mechanical\r\nand biological properties.\r\nMethods: 4-arm-PEG-DA was synthesized by graft copolymerization of linear PEG\r\n20,000 monomers, and characterized by IR1H NMR and 13C NMR; PEGylation of DAV\r\nwas achieved by the Michael addition reaction between propylene acyl and thiol, its\r\neffect was tested by uniaxial planar tensile testing, hematoxylin and eosin (HE) and\r\nscanning electron microscopy (SEM). Gly-Arg-Gly-Asp-Ser-Pro-Cys (GRGDSPC)\r\npeptides and vascular endothelial growth factor-165 (VEGF165) were conjugated onto\r\nDAV by branched PEG-DA (GRGDSPC-PEG-DAV-PEG-VEGF165).\r\nResults: Mechanical testing confirmed that PEG-cross-linking significantly enhanced\r\nthe tensile strength of DAV. Immunofluoresce confirmed the GRGDSPC peptides\r\nand VEGF165 were conjugated effectively onto DAV; the quantification of\r\nconjunction was completed roughly using spectrophotometry and ELISA. The\r\nhuman umbilical vein endothelial cells (HUVECs) grew and spread well on the\r\nGRGDSPC-PEG-DAV-PEG-VEGF165.\r\nConclusions: Therefore, PEGylation of DAV not only can improve the tensile strength\r\nof DAV, and can also mediate the conjugation of bioactive molecule (VEGF165 and\r\nGRGDSPC peptides) on DAV, which might be suitable for further development of\r\ntissue engineered heart valve.
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