纤维粘连蛋白与碱性成纤维细胞生长因子对成骨细胞黏附的协同刺激作用冯立曹晓建任永信【摘要】目的研究纤维粘连蛋白(fibronection,FN)与碱性成纤维细胞生长因子(basicfibroblastgrowthfactor,bFGF)单独或联合应用时对成骨细胞在生物衍生骨支架上黏附的影响,并探索其潜在机制。方法取第3代成骨细胞,以bFGF(011、1、10和100ng�ml)刺激,接种于FN(011、1、10和100Λg�ml)或多聚赖氨酸(Polylysine)修饰的生物衍生骨支架上,MTT法检测组间细胞黏附效率差异,电镜观察细胞密度与形态。GRGDS肽封闭后再次重复上述步骤。结果单独应用FN与bFGF均可增加成骨细胞在生物衍生骨支架上的黏附效率(P<0105),联合应用时,产生的效应显著增强,两者存在交互作用(P<0.01)。而Polylysine与bFGF无此交互作用(P>0105)。电镜观察发现,两者联合应用时细胞密度、铺展效果均优于单独应用。应用GRGDS肽后,FN联合bFGF产生的黏附促进作用被显著阻断。结论bFGF与FN对成骨细胞在生物衍生骨三维支架上的黏附存在协同刺激作用,细胞黏附产生这一效应的机制很可能是由RGD2整合素Α5Β1途径来介导的,需深入研究探讨。【关键词】骨组织工程纤维粘连蛋白碱性成纤维细胞生长因子生物衍生骨中图分类号:R318.17Q813.1文献标识码:ASYNERGISTICEFFECTOFFIBRONECTIONANDBASICFIBROBLASTGROWTHFACTORONOSTEOBLASTADHESIONEFFICIENCY�FENGLi,CAOXiaojian,RENYongxin.DepartmentofOrthopedics,FirstAffiliatedHospitalofNanjingMedicalUniversity,NanjingJiangsu,210029,P.R.China.E2mail:fengli19811010@163.comCorrespondingauthor:RENYongxin,E2mail:renyongxin@yahoo.com.cn【Abstract】ObjectiveToinvestigateeffectsofthebasicfibroblastgrowthfactor(bFGF)andfibronection(FN)ontheosteoblastadhesiononthebio2derivedbone.MethodsThethirdgenerationoftheosteoblastwastreatedwithbFGF0.1,1,10,and100ng�ml,respectively,andthenwasseededinthebio2derivedbone,whichhadbeenmodifiedwithFN0.1,1,10,and100Λg�ml,orPolylysine,respectively.ThecelladhesionwasmeasuredbytheMTTassay.Thecelldensityandthecellappearancewereobservedbythescanningelectronmicroscope.Theabove2men2tionedprocedureswererepeatedbyanapplicationoftheGRGDSpeptide.ResultsBothFNandbFGFcouldenhancetheosteoblastadhesionefficiencyonthebio2derivedbone(P<0.05).However,theosteoblastadhesionefficiencycouldbesignificantlystrengthenedbyacombineduseofFNandbFGF.FNandbFGFhadasignificantsynergisticeffectinstatistics(P<0.01),butPolylysineandbFGFhadnosuchsynergisticeffect(P>0105).ThecombineduseofFNandbFGFhadabettereffectonthecelldensityandthecellappearancethaneitherofthemwhenobservedwiththescanningelectronmicroscope.AdhesionefficiencygeneratedbythecombineduseofFNandbFGFwassignificantlyblockedbytheapplicationoftheGRGDSpeptide.ConclusionThecombineduseofFNandbFGFhasasignificantsynergisticeffectontheosteoblastadhesionefficiencyonthebio2derivedbone.ThiseffectisprobablymediatedbytheRGD2integrinΑ5Β1pathway.【Keywords】BonetissueengineeringFibronectionBasicfibroblastgrowthfactorBio2derivedboneFoundationitem:InnovatingFoundationofNanjingMedicalUniversity(CX2003016)成骨细胞为贴壁依赖细胞,细胞的黏附是其迁移、增殖、分化的基础,脱离了基质的贴壁细胞即很快停止于生长G1期或G0期[123]。目前骨组织工程的难题之一是支架材料与种子细胞的亲和性太差,导致细胞黏附率不足,影响细胞增殖分化和成骨能力。提高种子细胞基金项目:南京医科大学科技创新基金资助项目(CX2003016)作者单位:南京医科大学第一附属医院骨科(南京,210029)通讯作者:任永信,主治医师,研究方向:骨组织工程,E2mail:renyongxin@yahoo.com.cn与支架材料黏附能力,进而提高细胞接种效率,促进其增殖分化成骨,是解决这一问题的重要方法。生物衍生骨,具有天然的网状孔隙系统及纳米级羟基磷灰石排列结构,类似人体骨组织,符合...
