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bFGF基因转染BMSCs复合珊瑚骨构建下颌髁突的初步研究.docx

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bFGF基因转染BMSCs复合珊瑚骨构建下颌髁突的初步研究 摘要： 本研究旨在评估基础成纤维生长因子（bFGF）基因转染骨髓间充质干细胞（BMSCs）复合珊瑚骨在构建下颌髁突的应用效果。采用原代BMSCs，将其分为空白组、慢病毒转染bFGF基因组和普通病毒转染bFGF基因组。转染后细胞扩增，体外培养后与珊瑚骨进行复合，植入到Wistar大鼠髁突处。研究结果显示，基因转染后BMSCs增殖能力显著增强，其中慢病毒转染组增殖效果最佳；镜检发现慢病毒转染组珊瑚骨与周围组织更为紧密，并有较好的内生性修复效果。因此，bFGF基因转染BMSCs复合珊瑚骨构建下颌髁突具有潜在的临床应用前景。 关键词：bFGF基因；BMSCs；珊瑚骨；下颌髁突；复合；再生医学 Introduction Mandibularcondyle,asanimportantpartofthetemporomandibularjoint,playsakeyroleinthemovementofthemandibleandtheocclusalrelationship.Itsgrowthanddevelopmentarecloselyrelatedtothegrowthanddevelopmentofthecraniofacialskeleton.Thedestructionofthemandibularcondyleduetovariousreasonscanseriouslyaffectthepatient'squalityoflife.Therefore,thereconstructionofthemandibularcondyleisahottopicinthefieldofregenerativemedicine. Bonemarrowmesenchymalstemcells(BMSCs)havegoodpotentialforself-renewalanddifferentiationintomultiplecelltypes,includingosteoblasts,chondrocytes,andadipocytes.Atthesametime,theycanalsosecreteavarietyofcytokinestoplayanimportantroleintherepairofdamagedtissues.Inrecentyears,researchonstemcelltissueengineeringhasfocusedonthecombinationofstemcellsandbiomaterialstoconstructtissue-engineeredboneforclinicalapplication. Basicfibroblastgrowthfactor(bFGF)isamemberofthefibroblastgrowthfactorfamily,whichcanpromotetheproliferationanddifferentiationofosteoblastsandchondrocytesandpromotenewboneformation.Therefore,inthisstudy,weusedbFGFgene-transfectedBMSCscombinedwithcoralbonetoconstructthemandibularcondyle,andevaluateditsapplicabilityinvitro. MaterialsandMethods IsolationandcultureofBMSCs Bonemarrowsampleswereextractedfromsix4-week-oldWistarratsandsterilelyplacedinacentrifugetubecontainingphosphate-bufferedsaline(PBS).AfterwashingwithPBS,thecellsweredigestedwith0.25%trypsin-EDTAfor5minutes,andthentheculturesolutionwasaddedtoterminatethedigestion.Aftercentrifugation,thesupernatantwasdiscardedandtheprecipitationwasresuspendedandseededinacultureflaskcontainingDulbecco'smodifiedEagle'smedium(DMEM)supplementedwith10%fetalbovineserum(FBS).Thecellswer