脈沖電磁場(chǎng)誘導(dǎo)人骨髓間充質(zhì)干細(xì)胞向成骨細(xì)胞分化的研究

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1、中華物理醫(yī)學(xué)與康復(fù)雜志2005年1月第27卷第1期　ChinJPhysMedRehabil,January2005,Vol.27,No.1·5··基礎(chǔ)研究·脈沖電磁場(chǎng)誘導(dǎo)人骨髓間充質(zhì)干細(xì)胞向成骨細(xì)胞分化的研究趙敏　許建中　周強(qiáng)　王序全　朱灝　秦輝　單建林【摘　要】　目的　觀察脈沖電磁場(chǎng)(PEMFs)對(duì)人骨髓間充質(zhì)干細(xì)胞(hMSCs)向成骨細(xì)胞分化的影響。方法　分離并培養(yǎng)hMSCs,取第3代細(xì)胞,分為對(duì)照組與處理組,處理組細(xì)胞接受頻率12Hz、場(chǎng)強(qiáng)1.1mT的PEMFs刺激,每日8h;對(duì)照組細(xì)胞不接受PEMFs刺激。應(yīng)用透射電鏡觀察2組細(xì)胞超微結(jié)構(gòu),采用

2、堿性磷酸酶染色、骨鈣素免疫組織化學(xué)染色、四環(huán)素?zé)晒鈽?biāo)記等方法進(jìn)行觀測(cè)。結(jié)果　細(xì)胞培養(yǎng)5～6d即可傳代,經(jīng)PEMFs連續(xù)刺激3d后,hMSCs細(xì)胞形態(tài)發(fā)生變化,3周后聚集成細(xì)胞鈣化結(jié)節(jié)。透射電鏡下觀察顯示:細(xì)胞比較成熟,內(nèi)質(zhì)網(wǎng)擴(kuò)張,其堿性磷酸酶染色、骨鈣素免疫組織化學(xué)染色和四環(huán)素?zé)晒鈽?biāo)記均為陽(yáng)性。結(jié)論　hMSCs經(jīng)特定頻率和場(chǎng)強(qiáng)的PEMFs體外刺激后,可向成骨細(xì)胞分化,符合成骨細(xì)胞的形態(tài)學(xué)和生物學(xué)特性,為體外構(gòu)建組織工程化骨提供自體種子細(xì)胞。【關(guān)鍵詞】　脈沖電磁場(chǎng);　骨髓間充質(zhì)干細(xì)胞;　細(xì)胞分化;　成骨細(xì)胞Theeffectofpulsedelectro

3、magneticfieldsonthecellulardifferentiationfromhumanmesenchymalstemcellsto3osteoblastZHAOMin,XUJian2zhong,ZHOUQiang,WANGXu2quan,ZHUHao,QINHui,SHANJian23lin.OrthopedicResearchCenterofChinesePLA,DepartmentofOrthopedics,SouthwestHospital,the3rdMili2taryMedicalUniversity,Chongqing400

4、038,China【Abstract】ObjectiveToobservetheeffectofpulsedelectromagneticfields(PEMFs)onthecellulardiffer2entiationfromhumanmesenchymalstemcells(hMSCs)toosteoblast.MethodsWeisolatedandculturedthehMSCs,thendividedthepassagethreecellsintothecontrolgroupandthetreatmentgroup.Thetreatmen

5、tgroupwasstimulatedbythePEMFs(12Hzpulseburst,1.1mTpeak,8h/day).Transmissionelectronmicrograph,cyto2chemicalstainingforalkalinephosphatase(ALP),cytochemicalstainingforosteocalcinexpressionandtetracyclinefluorescencelabelwereappliedtomeasurethehMSCsdifferentiation.ResultsPEMFstrea

6、tmentacceleratedcel2lulardifferentiationsignificantly,hMSCscouldbepassedin6or5days,after3daysofexposure.Thecellularformwaschangedandtheygraduallycongregatedintoamineralizedmatrixin3weeks.Atransmissionelectronmicro2graphofthethirdpassageMSCsculturedwithexposuretoPEMFsshowedexpand

7、edendoplasmicreticulum.Cyto2chemicalstainingforALPandosteocalcinexpressionandtetracyclinefluorescencelabelwerepositive.ConclusionPEMFscouldacceleratethedifferentiationofhMSCstoosteoblast,whichservesastheautogenousseedingcellsforinvitroconstructionoftissueengineeringbone.【Keyword

8、s】Pulsedelectromagneticfields;Mesenchymalstemce