携带人胰岛素样生长因子ⅰ基因的重组腺病毒体外转导培养关节软骨细胞

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1、万方数据中国组织工程研究与临床康复第，2卷笫“勇咿2008—06一lO出版JournalofClinicalRehabilitativeTissueEnginee—ngResearchJune10,2008VoLl2,No．24携带人胰岛素样生长因子I基因的重组腺病毒体外转导培养关节软骨细胞江海良，夏青，魏振，黄爽，贾静Cultureofarticularchondrocytesthroughtransductionofadenovirus—humaninsulinlikegrowthfactor-Itransgene／nvitro．JiangHal-liang，XiaQing。Wei

2、Zhen，HuangShuang，JiaJingAbstractAIM：Insulinlikegrowthfactor-Iisalwaysregardedasoneofthekeygrowthfactorsincartilagegrowthandstability．ItcanpromotetypeIIcollagenandproteoglycanproductionbychondrocyte．Tllisstudyinvestigatedtheproductionofthesubstratecomponentofcartilagebyla'ansductionwithadenovirus

3、—humaninsulinlikegrowthfactor-I(Ad·hlGF-I)transgeneconstruct．METHODS：111eexperimentwasperformedattheLaboratoryofOrthopedics．FourthHospitalofDalianMedicalUniversityfromJanuary2005tOApril2007．ChondrocytesWereisolatedfrom3-week—oldrabbitkneeandfemoralheadandcultured．ThefirstpassagechondrocytesWerea

4、ssignedtotransductedgroupandnon—transductedgroupaftertheyweretransductedbyAd—hlGF—Iof500multiplicityofinfection．Thefirst．third．fifthandseventhpassagechondrocytes仃ansductedandnon—transductedwereassayedbyinvertedlightmicroscopytoobservetheirbiologicalcharacteristicchanges．ELISA，alkalinehydrolyseds

5、ampleassayandalicianbluetestwereperformedtoidentifytheexpressionandbiosynthesisofhydroxyproline，glycosaminoglycans(GAG)andIGF-Iprotein．RESULTS：Themicroscopicobservationshowedchondrocytesinthetransductedgroupproliferatedstably，andtriangle，andspindlecellsWerepredominantintheseventhpassage，accompan

6、iedbyfewlong·spindleandrootcells．Inaddition，thecellswereofclearappearanceandlargelucency．However，inthenon·transductedgroup，long—spindlecellswereobservedinthefourthpassage，andalmostallcellsexhibitedlong—spindleshapeinthefifthpassage．Wimculturetimeandpassageprolonging．theconcentrationofIGF-J．hydro

7、xyprolineandGAGintransductedchondrucytesculturemediumwassignificantlyhigherthanthes锄epassageceHsinnon—transductedgroup(P<0．05)．CONCLUSIoN：Theresultssuggest山atchondrocytestransductedwithAd—hIGF-IcouldproduceendogenousIGF-Iand