Study Of Biocompatibility Of Small Intestinal Submucosa Withendothelial Progenitor Cells Invitro

Objective：The prepared small intestinal submucosa (SIS) andendothelialprogenitorcells（EPCs）were co-cultured and biocompatibilitywas studied.The EPCs and EPCs transfected by Ad-VEGF165wereseeded on the surface of SIS,then we investigated theiradhension,proliferation.Method：EPCs was isolated from rat bone marrow by densitygradient centrifugation and cultured with EBM-2MV mediumcultivation.The recombinant adenovirus vector carrying the VEGF165gene was constructed by pAdEasy system.After infection with therecombinant Ad-VEGF165,EPCs were examined with a fluorescentmicroscopy and ELISA for their expression of VEGF,CCK-8assay wasemployed toassess the growthcharacter.EPCs,EPCs transfected by Ad-VEGF165and EPCs transfected byemptyvirus were respectivly culturedin virto with SIS.Then, EPCs wereexamined with scanning electron microscopy and CCK-8assay wasemployed to assess the growth character, ELISA for their expression ofVEGF andANG-1.Results：EPCs transfected byAd-VEGF165could highly expressedVEGF protein by ELISA.CCK-8confirmed EPCs had goodproliferationactivity on the surface of SIS,transfection group was significantly different from the other groups.It showed that EPCs adhered andproliferated well on the surface of SIS by SEM after3～5d.The proteingranules secreted on cellular surface were alsoshowed.Furthermore,ELISA measurement revealed that,cultured incombination with SIS, EPCs transfected by Ad-VEGF165secretedVEGF obvious difference with the other group and EPCs secretedANG-1without significantdifferencewith thegroups.Conclution：The SIS with EPCs have good biocompatibility fromtwo aspects of morphology and cell secretion,which was suitable as anatural scaffold material used in vascular tissueengineering.