摘要
目的合成具备MR显像和基因传输功能的纳米载体,探讨载体对间充质干细胞(MSCs)的基因传输功能及其联合生物发光成像(BLI)和MRI对MSCs双模态示踪的能力。方法合成三元共聚物聚乙二醇-聚天冬氨酸(聚乙烯亚胺)-超顺磁性氧化铁纳米颗粒载体(PAI/SPION);利用凝胶阻滞实验分析载体携带质粒(pDNA)的能力;电位及粒度测定仪测量载体复合pDNA后的粒径和电位;采用大鼠股骨骨髓分离培养MSCs;采用流式细胞仪、激光共聚焦显微镜评估PAI/SPION/pDNA对MSCs的基因转染效率;联合BLI和MRI双模态示踪MSCs。结果成功合成了载体PAI/SPION。氨基与质粒磷酸根的摩尔比(N/P)=3.0时,PAI/SPION可完全复合pDNA。N/P=12时,粒径趋于稳定,为(74.8±8.1)nm,电位为(12.2±1.5)mV,载体对MSCs的基因转染率为(71.2±2.3)%。激光共聚焦显微镜下,胞浆内可见大量表征PAI/SPION的绿色荧光和表征pDNA的红色荧光,且MSCs生物发光强度最高,T2*WI标准化信号强度最低。结论本研究成功合成了MRI可视的基因传输载体PAI/SPION;载体可高效传输pDNA至大鼠MSCs,且可成功联合BLI和MRI双模态示踪MSCs。
Objective To synthesize MRI-visible gene transfer nanocarrier and to assess the gene delivery ability of nanocarrier to mesenchymal stem cells(MSCs)and monitored the MSCs using bioluminescence imaging(BLI)and MRI.Methods The terpolymer nanocarrier poly(ethylene glycol)-block-poly(L-aspartic acid)-grafted polyethylenimine functionalized with superparamagnetic iron oxide nanoparticles(PAI/SPION)was synthesized.The degree of binding between PAI/SPION and pDNA was determined with agarose gel electrophoresis.The Zeta potential and particle size of polymer vectors were measured with a Zeta-Plus instrument.MSCs were isolated and cultured from the femurs of the rats.The gene transfection efficiency of PAI/SPION/pDNA to MSCs was evaluated by flow cytometry and laser confocal microcopy.And the BLI and MRI were combined to monitor MSCs.Results PAI/SPION vectors were synthesized successfully.PAI/SPION could condense pDNA entirely at a nitrogen of non-viral vector/phosphorus of plasmid(N/P)ratio of 3.0.The particle size of PAI/SPION/pDNA attained a constant size of(74.8±8.1)nm at a N/P ratio of 12,and the Zeta potential reached(12.2±1.5)mV,the transfection efficiency of PAI/SPION/pDNA to MSCs was(71.2±2.3)%.The MSCs showed significantly stronger PAI/SPION(green)and pDNA(red)fluorescence under the laser confocal microcopy,the bioluminescence light intensity of the MSCs attained the highest values and the normalized MR T2*WI signal intensity attained the lowest values.ConclusionThe study successfully synthesizes an MRI-visible non-viral gene vector PAI/SPION,which possesses a higher transfection efficiency in transferring pDNA into MSCs and can dually monitor the MSCs using BLI and MRI.
作者
吴春
姜在波
朱康顺
黄明声
关守海
李征然
钱结胜
李名安
单鸿
WU Chun J IANG Zaibo ZHU Kangshun HUANG Mingsheng GUAN Shouhai LI Zhengran QIAN Jiesheng LIMing'an SHAN Hong"(Department of Radiology, the Third Affiliated Hospital, Insitute of Interventional Radiology, Sun Yat-sen University, Guangzhou 510630, China)
出处
《中国医学影像技术》
CSCD
北大核心
2017年第2期171-176,共6页
Chinese Journal of Medical Imaging Technology
基金
广东省自然科学基金博士启动项目(2015A030310171)
关键词
间充质干细胞
基因
生物发光
磁共振成像
Mesenchymal stromal cells
Genes
Bioluminescence
Magnetic resonance imaging