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!"#$10005404 2014 12127306 !" #$%&'()%*+,-./012345678-9:;<= ! "# $% &! '( )( * 401147!"!"#$%&'(#&)'(*+,-.#&!" /!0123>?%& 456789:;<8=>?@ABCDE SD%FGHIJKLMNratmesenchymalstemcells rMSCs OPQRSTGUVWXYZ'( V[\]Q rMSCs TGU^_VWS`aMNbcdefWgVhi j89 30 70 100nmX TiO ;<8klmnop=>qrstuvwxy=>z{|}MTT g~vMNxy= >OPSQRnopMNxy=>qrMN:89 30 70 100nmX TiO ;<8= >TGUVW}}WSWRealtimePCR~v^_ 14d Runx2 S OSX= )* V[X rMSCs TGUVWX ¡CD29 d}MN¢ 99.65%klmnop£¤xy=>89¥ ¦§¨©ª89¢ 70nmX TiO ;<8xy=>z|}«¬<005 ); MTT ?-® 70nm¤ rMSCs XOPQR¥ «¯°¤±Q²<005 ); 30nm¤TG^_ 7d }}¥²³¯°¤<005 )。30nm¤TG^ _ 21d =>W²³¯°¤<005 )。30nm¤TG^_ 14d Runx2 S OSX=²³¯´¤<005 )。 )+ ,µ>:¶·6789X TiO ;<8A rMSCs X¸¹£6¶7rMSCs 70nm=>QR ¡ºOP}« ¬» 30nm=>XTGUVW ¡«¼@ABTiO ;<8GHIJKLMNTGUVW CDE9FGR318.08 R329.28 R331.22     HIJKLMNOP!"#$%&'(#&)½¾$4¿ÀÁ2011 ); !"/Â-Ã>ÄÅÆ20122121 ); !"/$ÇÈÉ$&Á CSTC2012jjA0178 ); ²Ê&˽¾ &$0ÌÅ$4ÁÍÎÏÐÑÒÓÔ20125503120009 QRSTÕ ÖEmail ysdentist@hotmail.com UVWXhttp //www.cnki.net/kcms/detail/51.1095.R.20140114.1843.001.html 20140114 EffectsofdifferentdiameterTiO nanotubesonproliferationandosteogenicdiffer entiationofratmesenchymalstemcells WangMing YangSheng DengFeng WangLu ZhangYi ZhangHeChongqingKeyLaboratoryforOralDiseasesand BiomedicalSciences CollegeofStomatology ChongqingMedicalUniversity Chongqing 401147 China Abstract Objective ToinvestigatetheeffectsofdifferentdiameterTiO nanotubespreparedby anodizationontheadhesion proliferationandosteogenicdifferentiationofSpragueDawleyratmesenchymal stemcells rMSCs Methods TheisolatedrMSCswerecharacterizedbyosteogenicdifferentiationandflow cytometry.TiO nanotubeshavingdiameterof30 70and100nmwerefabricatedbyanodizationwithdifferent voltages.Thetopographyandhydrophilicityofthenanotubeswereaccessedwithscanningelectronmicroscopy SEM andcontactangletest respectively.MTTassaywasappliedtoevaluatethecelladhesionandprolifera tionondifferentsubstrates.Cellmorphologieswereobservedbyconfocalmicroscopy.Theactivityofalkaline phosphataseandmineralizationofrMSCsonplanetitanium andTiO nanotubesdiameter=30 70and 100nm werealsodetectedduringosteogenicdifferentiation.TheexpressionofRunx2andOSXwasanalyzed byrealtimePCRin14dafterosteogenicdifferentiation. Results TheosteogenicdifferentiationofrMSCswas successfullyinduced andCD29positivecellswere99.65%.TiO nanotubesofdifferentdiametersweregenera ted.TheTiO nanotubesof70nmdiameterweremorehydrophilic <005 andexhibitedbettercelladhe sionandproliferationcomparedwithothersubstrates <005 .ThecellsculturedontheTiO nanotubesof 30nmdiameterdisplayedmuchhigheractivityofalkalinephosphatasethanthoseonothersubstratesin7dafter osteogenicinduction <005 ), highermineralizationin21dafterosteogenicinduction <005 andmore × 36 Ø× 12 Ù 2014 Ú Û 30 Ü × Ý Þ # % & & ß J Third Mil Med Univ Vol.36 No.12 Jun.30 2014

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EffectsofdifferentdiameterTiO2nanotubesonproliferationandosteogenicdifferentiationofratmesenchymalstemcellsWangMing,YangSheng,DengFeng,WangLu,ZhangYi,ZhangHe(ChongqingKeyLaboratoryforOralDiseasesandBiomedicalSciences,CollegeofStomatology,ChongqingMedicalUniversity,Chongqing,401147,China)

  [Abstract] Objective ToinvestigatetheeffectsofdifferentdiameterTiO2nanotubespreparedbyanodizationontheadhesion,proliferationandosteogenicdifferentiationofSpragueDawleyratmesenchymalstemcells(rMSCs).Methods TheisolatedrMSCswerecharacterizedbyosteogenicdifferentiationandflowcytometry.TiO2nanotubeshavingdiameterof30,70and100nmwerefabricatedbyanodizationwithdifferentvoltages.Thetopographyandhydrophilicityofthenanotubeswereaccessedwithscanningelectronmicroscopy(SEM)andcontactangletest,respectively.MTTassaywasappliedtoevaluatethecelladhesionandproliferationondifferentsubstrates.Cellmorphologieswereobservedbyconfocalmicroscopy.TheactivityofalkalinephosphataseandmineralizationofrMSCsonplanetitaniumandTiO2 nanotubes(diameter=30,70and100nm)werealsodetectedduringosteogenicdifferentiation.TheexpressionofRunx2andOSXwasanalyzedbyrealtimePCRin14dafterosteogenicdifferentiation.Results TheosteogenicdifferentiationofrMSCswassuccessfullyinduced,andCD29positivecellswere99.65%.TiO2nanotubesofdifferentdiametersweregenerated.TheTiO2nanotubesof70nmdiameterweremorehydrophilic(P<005)andexhibitedbettercelladhesionandproliferationcomparedwithothersubstrates(P<005).ThecellsculturedontheTiO2nanotubesof30nmdiameterdisplayedmuchhigheractivityofalkalinephosphatasethanthoseonothersubstratesin7dafterosteogenicinduction(P<005),highermineralizationin21dafterosteogenicinduction(P<005)andmore

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Runx2andOSXgeneexpressionin14dafterosteogenicinduction(P<005).Conclusion TiO2nanotubeswithdifferentdiametershavedifferentinfluencesonrMSCsascomparedtoplanetitanium.TheTiO2nanotubesof70nmdiameteraremoreadvantageousforadhesionandproliferationofrMSCs,whileTiO2nanotubesof30nmdiameterfacilitateosteogenicdifferentiation.  [Keywords] TiO2nanotubes;mesenchymalstemcells;osteogenicdifferentiation

SupportedbytheDoctoralStartingFundofCollegeofStomatologyofChongqingMedicalUniversity(2011),theGeneralProgramofChongqingMunicipalHealthBureau(20122121),theNaturalScienceFoundotionofChongqing(CSTC2012jjA0178)andtheProjectforNewFeachersFromHighEducationalDoctorialDisciplineScienceFoundation(20125503120009).Correspondingauthor:ZhangHe,Email:ysdentist@hotmail.com

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