压力与牙周膜细胞对体外诱导脐血单核细胞分化为破骨样细胞的影响

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1、压力与牙周膜细胞对体外诱导脐血单核细胞分化为破骨样细胞的影响【摘要】　　目的研究持续静压力和人牙周膜细胞对体外诱导脐血单核细胞分化为破骨样细胞的影响，初步探讨静压力和牙周膜细胞在正畸性骨改建中的作用机制。方法密度梯度离心法分离脐血，收集单个核细胞。用组织块酶消化法培养获得牙周膜细胞。建立transwell共培养系统，下层接种脐血单个核细胞，上层接种牙周膜细胞。A组为单核细胞与牙周膜细胞共培养加力；B组为单核细胞培养加力，另设不加力对照组A’、B’组。采用倒置相差显微镜观察及TRAP染色，骨磨片染色等方法检测破骨样细胞的形成及

2、功能。结果A组下层细胞在加力后第2天出现细胞融合现象，3d时可见明显的多核破骨样细胞，TRAP染色阳性，骨磨片染色可见典型的骨吸收陷窝。其他组仅有极少量多核细胞形成，未见骨吸收陷窝。结论静压力作用下，牙周膜细胞可以刺激脐血单核细胞分化为破骨样细胞。【关键词】脐血单核细胞牙周膜细胞持续静压力破骨样细胞　　ABSTRACT:ObjectiveTostudytheeffectofcontinuouslycompressivepressure(CCP)andhumanperiodontalligamentcells(HPDLCs)o

3、nthedifferentiationofosteoclastlikecells(OLC)inducedfromumbilicalcordbloodcellsinvitro,andtoinvestigatetheroleofcontinuouslycompressivepressureand15humanperiodontalligamentcellsinalveolarbonerebuildingduringorthordontictoothmovement.MethodsMononuclearedcellsofumbi

4、licalcordblood(HCMNCs)wereseparatedbydensitygradientcentrifugation,HPDLCswereisolatedfromhumanperiodontalligamentbyexplantingenzymaticdigestionwithtrypsinandcollagenase.WealsoestablishedtranswellcoculturesystemwithHCMNCsinthelowerlayerandHPDLCsintheupperlayer.Grou

5、pA:HCMNCsandHPDLCswerecoculturedwith150kPaCCPfor1.5hoursonthemodel.GroupB:onlyHCMNCswereculturedwiththesameCCPasGroupA.GroupsAandBweretherespectivecontrolgroupofGroupsAandBwithnoCCPexerted.Thecellappearancewasobservedunderthephasecontrastmicroscope,anditsidentif

6、icationwasperformedbyhistochemicalanalysisoftartraterresistantacidphosphatase(TRAP).Thecapacityofboneresorptionofthecellwasassessedbylacunaeformingabilityonboneslice.ResultsHCMNCsinGroupAbegantofuseonthe2ndday,MorepositivemultinucleatedcellscouldbeseenwithTRAPstai

7、ningandcorticalbonepitformationonthe3rdday.Onlyafewmultinucleatedcellsformedintheothergroups,withnocorticalbonepitformation.ConclusionHCMNCscanfuseintomultinucleatedOLCunderCCPwiththeinductionofHPDLCs.15　　KEYWORDS:mononuclearedcellofumbilicalcordblood;humanperiodon

8、talligamentcell;continuouslycompressivepressure;osteoclastlikecell　　正畸牙移动是一个复杂的机械生物学反应过程，牙周组织感应力学刺激后所发生的组织改建是牙齿移动的基础。破骨细胞(osteoclast,OC)是骨吸收的最重要