人骨形成蛋白7基因修饰的兔骨髓基质干细胞异位成骨实验

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1、人骨形成蛋白7基因修饰的兔骨髓基质干细胞异位成骨实验作者：闫露，罗二平，鱼兵，栗艳，沈敏，范清宇【摘要】［目的］通过人骨形成蛋白7(humanbonemorphogeneticprotein7，BMP7)基因修饰的兔骨髓基质干细胞(mesenchymalstemcells,MSCs)与聚乳酸-羟基乙酸(polycleclidecoplycoclide,PLGA)支架复合，进行自体皮下异位成骨实验，探索BMP-7基因治疗与组织工程技术相结合的可行性。［方法］PLGA与腺病毒Ad-BMP-7感染后的兔MSCs共培养，电镜观察了解细胞在PLG

2、A上黏附、生长和合成分泌骨基质成分的情况；将转染基因的细胞和未转染基因的细胞分别与PLGA支架复合，构建组织工程化骨并植入体内，通过组织观察新骨形成情况。［结果］制备得到的PLGA为疏松多孔的网格样结构，具有一定的韧性和强度。将腺病毒感染的骨髓基质干细胞接种于修剪成一定形状的PLGA上，经电镜观察可见细胞贴附于材料表面和孔隙内并增殖。异位成骨实验证实空白对照PLGA孔隙内无新骨形成，对照细胞组内有部分新生骨形成，而AdBMP7实验组形成的新生骨组织面积更大(P<0.05)。［结论］应用hBMP7基因修饰的MSCs作为骨组织工程的

3、种子细胞，可望取得更强的成骨能力。【关键词】基因治疗；骨形成蛋白；组织工程12　　Abstract：［Objective］hBMP7genetherapycombinedwithtissueengineeringtechniqueswasexploredtoimproveosteogenesisinanectopicboneformationmodelinrabbits.［Method］PLGA(withdiameterof150～300μmandporosityof90%)waspreparedandautologousbonemar

4、rowstromalcells(bMSCs)infectedbyadenoviruscarringhBMP7gene(AdBMP7)wereseededontoit.Scanningelectronicmicroscopingwereusedtoobservecellmatrixinteraction.ThecellsinfectedandnoninfectedwerecombinedwithPLGAataconcentrationof5×106cells/mltoconstructtissueengineeredbonesre

5、spectively.Theywerefurtherimplantedinto8rabbitssubcutaneouslyusingPLGAasblankcontrol(8implantspergroup).Sixweeksaftersurgery,theimplantswereevaluatedwithhistologicalstainingandcomputerizedanalysisofnewboneformation.［Result］PLGApreparedwasagridlikeporousstructure,withdefin

6、iteductilityandstrength,andcouldbetrimmedintodifferentmodels.AdenovirusinfectedMSCsseededontoPLGAshowedhighlevelofproliferation,andmasssynthesisofcellmatrixwasobservedwithelectronicmicroscoping.Intheectopicboneformationmodel,PLGAalonecouldnotinducenewboneformation.Thenewb

7、oneareaformedinthenoninfected12MSCsgroupwas28.6±6.1%,whileAdBMP7groupwas57.5±5.7%,whichwashigherthanthatoftheformersignificantly(P<0.05).［Conclusion］hBMP7genemodifiedMSCscouldenhanceectopicnewboneformationinrabbits.TheseresultsindicatedthattheinfectionofMSCswithhBMP

8、7adenovirusmightbesuitableforbonetissueengineeringapplications.　　Keywords：genetherapy；bo