MRI对自体骨髓间质干细胞移植于缺血心肌后动态监测价值的实验研究
MRI대자체골수간질간세포이식우결혈심기후동태감측개치적실험연구
Dynamic MRI of ferumoxide-labeled bone mesenchymal stem cells after transplantation in infarcted myocardium
  • 万方数据
    中华放射学杂志 중화방사학잡지
    Chinese Journal of Radiology
    2009年 3期 305-310 ,共6页

    刘琼%赵世华%陆敏杰%蒋世良%闫朝武%张岩%孟亮%唐跃%孟宪敏%魏英杰%王青峙

    류경%조세화%륙민걸%장세량%염조무%장암%맹량%당약%맹헌민%위영걸%왕청치

    磁共振成像%问质十细胞%心肌梗死 磁共振成像%問質十細胞%心肌梗死
    자공진성상%문질십세포%심기경사
    Magnetic resonance imaging%Mesenchymal stem cell%Myocardial infarction
    目的 评价MRI对干细胞移植于缺血心肌后的动态监测价值.方法 中华小型猪6头,抽取髂骨骨髓并制备自体骨髓间质干细胞(mesenchymal stem cells,MSCs),以注射用超顺磁性氧化铁颗粒(SPIO)及4',6.二脒基-2.苯基吲哚(DAPI)标记细胞并进行标记率检测.开胸结扎冠状动脉左前降支制备小型猪心肌梗死模型.模型建立后14 d,MR延迟增强成像(DE-MRI)检测梗死面积,之后行第2次开胸,于每只动物心肌梗死周边区和正常心肌区直视下经心外膜各注射标记的MSCs 2个点,同时分别于各区注射培养基2个点作为对照点.细胞移植后24 h及21 d,快速梯度回波序列(F'GRE)T2*像检测十细胞移植点低信号区的面积及信号强度.T2*信号减低区范围的测量由FGRE面积法实现,其信号减低的程度以正常心肌区和该区的T2*值之差与正常心肌区T2*值的百分比表示.病理组织学检查心肌细胞形态、瘢痕形成、毛细血管密度及干细胞分布情况.不同时间点MSCs注射点低信号区面积及T2*信号强度的比较采用重复测量方差分析及配对t检验.干细胞注射点与对照点毛细血管密度的比较、梗死周边区及正常心肌区MSCs注射点信号衰减幅度和DAPI阳性细胞密度的比较采用成组资料t检验.结果 DAPI及SPIO对MSCs的标记率均达100%.心肌梗死模型建立后第14天,小型猪心肌梗死面积为(33.6±8.9)%.细胞移植后24 h,标记MSCs注射点于MRI显示为边界清晰的卵圆形T2*低信号区,梗死周边区与正常心肌区MSCs注射点T2*低信号区的信号强度[分别为(67.00±5.48)%、(61.92±7.76)%,t=1.65,P=0.1158)]及面积[分别为(0.56±0.24)、(0.52±0.25)cm2,t=0.39,P=0.7044.)]差异均无统计学意义.移植后3周,T2*低信号区与正常心肌区的对比程度均较前下降,梗死周边区减低至(40.12±5.93)%(t=9.53,P＜0.01);正常心肌区减低为(46.92±6.25)%(t=11.03,P＜0.01).梗死边缘区T2*减低的程度大于正常心肌区MSCs注射点,且2组间信号强度减弱幅度分别为(26.88±7.27)%和(15.00±4.51)%,差异有统计学意义(F:20.08,P=0.0003).正常心肌区MSCs注射点心肌组织中荧光标记的细胞核分布密度高于梗死边缘区.MSCs注射点[分别为(106±25)和(143±31)个/高倍镜(f=-2.47,P=0.0293)].梗死边缘区MSCs注射点组织中毛细血管分布密度高于该区对照点[分别为(13.4±4.0)和(9.4±3.1)个/高倍镜,f=2.49,P=0.0229].MSCs移植于心肌组织中3周后,普鲁十蓝染色阳性的铁颗粒仅位于部分移植MSCs细胞核周围.结论 MRI可在一定时间内实现对移植干细胞的示踪并反映移植MSCs在心肌局部的数量变化趋势,但是对于移植细胞的半定量监测存在局限性.移植干细胞所在的微环境是影响其牛存时间的重要因素之一.
    목적 평개MRI대간세포이식우결혈심기후적동태감측개치.방법 중화소형저6두,추취가골골수병제비자체골수간질간세포(mesenchymal stem cells,MSCs),이주사용초순자성양화철과립(SPIO)급4',6.이미기-2.분기신타(DAPI)표기세포병진행표기솔검측.개흉결찰관상동맥좌전강지제비소형저심기경사모형.모형건립후14 d,MR연지증강성상(DE-MRI)검측경사면적,지후행제2차개흉,우매지동물심기경사주변구화정상심기구직시하경심외막각주사표기적MSCs 2개점,동시분별우각구주사배양기2개점작위대조점.세포이식후24 h급21 d,쾌속제도회파서렬(F'GRE)T2*상검측십세포이식점저신호구적면적급신호강도.T2*신호감저구범위적측량유FGRE면적법실현,기신호감저적정도이정상심기구화해구적T2*치지차여정상심기구T2*치적백분비표시.병리조직학검사심기세포형태、반흔형성、모세혈관밀도급간세포분포정황.불동시간점MSCs주사점저신호구면적급T2*신호강도적비교채용중복측량방차분석급배대t검험.간세포주사점여대조점모세혈관밀도적비교、경사주변구급정상심기구MSCs주사점신호쇠감폭도화DAPI양성세포밀도적비교채용성조자료t검험.결과 DAPI급SPIO대MSCs적표기솔균체100%.심기경사모형건립후제14천,소형저심기경사면적위(33.6±8.9)%.세포이식후24 h,표기MSCs주사점우MRI현시위변계청석적란원형T2*저신호구,경사주변구여정상심기구MSCs주사점T2*저신호구적신호강도[분별위(67.00±5.48)%、(61.92±7.76)%,t=1.65,P=0.1158)]급면적[분별위(0.56±0.24)、(0.52±0.25)cm2,t=0.39,P=0.7044.)]차이균무통계학의의.이식후3주,T2*저신호구여정상심기구적대비정도균교전하강,경사주변구감저지(40.12±5.93)%(t=9.53,P＜0.01);정상심기구감저위(46.92±6.25)%(t=11.03,P＜0.01).경사변연구T2*감저적정도대우정상심기구MSCs주사점,차2조간신호강도감약폭도분별위(26.88±7.27)%화(15.00±4.51)%,차이유통계학의의(F:20.08,P=0.0003).정상심기구MSCs주사점심기조직중형광표기적세포핵분포밀도고우경사변연구.MSCs주사점[분별위(106±25)화(143±31)개/고배경(f=-2.47,P=0.0293)].경사변연구MSCs주사점조직중모세혈관분포밀도고우해구대조점[분별위(13.4±4.0)화(9.4±3.1)개/고배경,f=2.49,P=0.0229].MSCs이식우심기조직중3주후,보로십람염색양성적철과립부위우부분이식MSCs세포핵주위.결론 MRI가재일정시간내실현대이식간세포적시종병반영이식MSCs재심기국부적수량변화추세,단시대우이식세포적반정량감측존재국한성.이식간세포소재적미배경시영향기우존시간적중요인소지일.
    Objective To investigate the ability of magnetic resonance imaging (MRI) in tracking magnetically labeled mesenchymal stem cells (MR-MSCs) in a swine myocardial infarction (MI) model.Methods Adult Chinese mini-pigs (n = 6) were subjected to open-chest experimental MI operation.Their antegeneic bone marrow-derived mesenchymal stem cells ( MSCs ) was cultured and doubly labeled with ferumoxides and DAPL On the 14 th day after MSCs transplantation, the size and location of the myocardial infarction were assessed by using delayed-enhancement MRI (DE-MRI). Then the labeled MSCs were injected intramyocardially into peri-infarct zone and normal myocardium. At 24 hrs and 3 weeks after injection, the contrast and the volume of the MR-MSCs hypointense lesion from the MR images were acquired, and the contrast was determined using the difference in signal intensity between the hypointense and normal myocardium divided by signal intensity of the normal region.After humane euthanasia, the heart was excised and histology corresponding to MRI slices that demonstrated MR-MSCs lesions was performed.Repeated-measures ANOVA and a paired t test were used for comparison of the contrast and the volume of the MR-MSCs hypointense lesion at different time points. Comparisons between independent groups were performed with the standard Student t test.Results The labeling efficiency of ferumoxides and DAPI was 100% . On the 14 th day after the MI operation, the average percentage of infracted myocardial area was (33.6±8.9)% .Twenty- four hours after MSCs transplantation, MSCs injection sites appeared as ovoid hypointensive lesions with sharp border on T2 * images. At 24 h after injection, the signal contrast [(67.00±5.48)% vs (61.92 ±7.76)%,t = 1.65,P =0.1158] and the size [(0.56 ±0.24) cm2 vs (0.52 ± 0.25 ) cm2, t = 0.39, P = 0.7044 ] of the lesions showed no statistical difference between the peri infarct zone and the normal myocardium.At 3 weeks after injection, the signal contrast decreased and the size diminished both in the peri-infarct zone and in the normal myocardium. Moreover, the contrast of the lesions in peri-infarct zone decreased more significantly than that in normal myocardium [(26.88 +7.27)%vs (15.00 :t:4.51)%, F =20.08, P =0.0003].Post mortem analysis found the fluorescontly labeled MSCs demonstrated on histological sections.There were much more dense fluorescently labled MSCs per high power fields at injection sites of normal myocardium than at injection sites of peri-infarct zone [ (106 ±25 )/HPF vs ( 143 ± 31 )/HPF, t = - 2.47, P = 0.0293 ].In MSCs injection sites of the peri-infarct zone,the capillary density was significantly higher than that in control sites [ (13.4 ± 4.0 )/HPF vs (9.4 ±3.1 )/HPF, t = 2.49, P = 0.0229].At 3 weeks after injection, ferumoxide was contained within partial original MSCs.Conclusion Magnetic resonance imaging of MSCs is a feasible method for the in vivo tracking of transplanted stem cells and could reflect the tendency of the local stem cell quantity, but there still has limitation for the semi-quantitation of the transplanted stem cells.
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