CAJ | 학술논문

磁共振成像/方法%内皮,血管/细胞学%脐静脉/细胞学%细胞,培养的%染色与标记%显微镜检查,电子,扫描透射%超顺磁性氧化铁%体外试验%荧光染色磁共振成像/方法%內皮,血管/細胞學%臍靜脈/細胞學%細胞,培養的%染色與標記%顯微鏡檢查,電子,掃描透射%超順磁性氧化鐵%體外試驗%熒光染色
자공진성상/방법%내피,혈관/세포학%제정맥/세포학%세포,배양적%염색여표기%현미경검사,전자,소묘투사%초순자성양화철%체외시험%형광염색
Magnetic resonance imaging/methods%Endothelium,vascular/cytol%Umbilical veins/cytol%Cells,cultured%Staining and labeling%Microscopy,electron,scanning transmission%Superparamagnetic iron oxide%In vitro test%Fluorescence

目的:研究磁共振对比剂超顺磁性氧化铁(SPIO)对血管内皮细胞生物学影响及磁共振成像的可能性,为SPIO对血管内皮细胞靶向性成像提供依据.方法:采用经典共沉淀法,合成柠檬酸包被的SPIO.实验组为浓度分别为0.01、0.05、0.10、0.15 mg/ml的SPIO,共同孵育人脐静脉血管内皮细胞,对照组为细胞在不含SPIO的培养液中孵育.采用普鲁士蓝染色,评价血管内皮细胞对SPIO的摄取情况.采用Calcein-AM法检测细胞活性的变化.通过免疫荧光染色对血管内皮细胞的肌动蛋白、微管蛋白进行荧光染色,评价血管内皮细胞骨架的变化.采用免疫荧光染色的方法,对局部粘着斑蛋白激酶(FAK)进行荧光染色,评价细胞迁移、粘附能力的变化.通过透射电镜观察SPIO在细胞内的分布及细胞内细胞器的变化.采用原子吸收分光光度计检测细胞内SPIO的含量.通过Philips 3.0T 磁共振,对细胞琼脂凝胶细胞悬液进行磁共振扫描,了解磁共振扫描时信号的变化.结果:血管内皮细胞按浓度依赖式摄取SPIO.与对照组相比随SPIO孵育浓度的增加,SPIO对细胞的活性的毒性增加.与对照组相比细胞的肌动蛋白、微管蛋白纤维紊乱,FAK颗粒变粗,分布稀疏.SPIO主要位于胞质的溶酶体内,随孵育浓度的增加,溶酶体的数量增多,并出现大小不一空泡样结构.当SPIO浓度为0.15 mg/ml时,细胞内铁含量为(55.86±9.935)pg.磁共振扫描图像显示与SPIO共同孵育的细胞,信号出现明显的降低.结论:摄取SPIO的血管内皮细胞可以被MR检测到,但SPIO对血管内皮细胞的活性、骨架、粘附、迁移造成了一定影响,而这些影响与SPIO浓度有明显的依赖性.
목적:연구자공진대비제초순자성양화철(SPIO)대혈관내피세포생물학영향급자공진성상적가능성,위SPIO대혈관내피세포파향성성상제공의거.방법:채용경전공침정법,합성저몽산포피적SPIO.실험조위농도분별위0.01、0.05、0.10、0.15 mg/ml적SPIO,공동부육인제정맥혈관내피세포,대조조위세포재불함SPIO적배양액중부육.채용보로사람염색,평개혈관내피세포대SPIO적섭취정황.채용Calcein-AM법검측세포활성적변화.통과면역형광염색대혈관내피세포적기동단백、미관단백진행형광염색,평개혈관내피세포골가적변화.채용면역형광염색적방법,대국부점착반단백격매(FAK)진행형광염색,평개세포천이、점부능력적변화.통과투사전경관찰SPIO재세포내적분포급세포내세포기적변화.채용원자흡수분광광도계검측세포내SPIO적함량.통과Philips 3.0T 자공진,대세포경지응효세포현액진행자공진소묘,료해자공진소묘시신호적변화.결과:혈관내피세포안농도의뢰식섭취SPIO.여대조조상비수SPIO부육농도적증가,SPIO대세포적활성적독성증가.여대조조상비세포적기동단백、미관단백섬유문란,FAK과립변조,분포희소.SPIO주요위우포질적용매체내,수부육농도적증가,용매체적수량증다,병출현대소불일공포양결구.당SPIO농도위0.15 mg/ml시,세포내철함량위(55.86±9.935)pg.자공진소묘도상현시여SPIO공동부육적세포,신호출현명현적강저.결론:섭취SPIO적혈관내피세포가이피MR검측도,단SPIO대혈관내피세포적활성、골가、점부、천이조성료일정영향,이저사영향여SPIO농도유명현적의뢰성.
Objective: To investigate the magnetic resonance (MR) signal changes of superparamagnetic iron oxide (SPIO) and its biological effects on endothelial cells. Methods: The citric-acid coated SPIO was synthesized by co-precipitation method.The human umbilical vein endothelial cells (HUVECs) were incubated with SPIO for 24 h in culture medium at iron concentration of 0.01,0.05,0.10,0.15 mg/ml (experimental groups),and the cells incubated without SPIO served as control groups.The uptake efficiency of intracellular iron was measured by Prussian blue staining,and the cell viability was monitored by Calcein-AM method.The cell cytoskeleton (F-actin and tubulin),adherence and migration capacity were measured by immunofluorescence staining.The iron oxide nanoparticles distribution and the cellular organelle change were monitored by transmission electron microscopy (TEM).Quantification of particle uptake was measured by atomic absorption spectrometry.The MR signal of endothelial cells after labeling was monitored by Philips 3.0 T MR scanner. Results: SPIO was uptaken by HUVECs in a concentration-dependence manner.Compared with the control group,cell viability was decreased along with the increase of iron concentration.Compared with the control group,the cell cytoskeleton was markedly disorganized and the FAK spot was bigger and sparser.The nanoparticles were mainly existed in lysosomes,and the higher concentration of SPIO,the more lysosomes and vacuoles presented in the cells.The iron content per cell was (55.86±9.935)pg when the SPIO concentration was 0.15 mg/ml.The MR image showed that the cells labeled with SPIO resulted in the decrease of MR signal. Conclusion: The cells labeled with SPIO can be detected by MR.The cell viability,cytoskeleton,adherence and migration capacity of HUVECs are affected by citric-acid coated SPIO in a concentration-dependent manner.