Li You-Wei, Chen Zheng-Guang, Zhao Zhou-She, Li Hong-Li, Wang Ji-Chen, Zhang Zong-Ming
You-Wei Li, Department of Radiology, Beijing Chuiyangliu Hospital Affiliated to School of Medicine, Tsinghua University, Beijing 100022, China.
World J Gastroenterol. 2015 Apr 14;21(14):4275-83. doi: 10.3748/wjg.v21.i14.4275.
To prepare the specific magnetic resonance (MR) probes for detection of hepatocellular carcinoma (HCC) using one-pot method.
The carboxylated dextran-coated nanoparticles were conjugated with anti-α-fetoprotein (anti-AFP) or anti-glypican 3 (anti-GPC3) antibodies through 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride/N-hydroxysuccinimide (EDC/NHS)-mediated reaction to synthesize the probes. The physical and chemical properties of the probes were determined by transmission electron microscopy (TEM) and dynamic light scattering, and the relaxivity was compared to uncombined ultrasmall superparamagnetic iron oxide nanoparticles (USPIONs) using a 1.5T clinical MR scanner. The binding efficiency of the antibodies to nanoparticles was measured with an ultraviolet-visible spectrophotometer. In addition, the probes were incubated with targetable cells in vitro.
The superparamagnetic MR probes (anti-GPC3-USPION probe and anti-AFP-USPION probe) were synthesized using one-pot method. Their mean hydrodynamic diameter was 47 nm with a broader slight size distribution. The coupling efficiency of carboxylated dextran-coated ultrasmall superparamagnetic iron oxide (USPIO) with anti-GPC3 or anti-AFP antibody was 15.9% and 88.8%, respectively. Each of the USPIO nanoparticles may bind 3 GPC3 antibodies or 12 AFP antibodies. The statistical analysis showed no significance (P > 0.05) in shortening the T1 and T2 values when comparing the USPIO-AFP or USPIO-GPC3 to USPIO. Analysis of TEM images revealed that anti-GPC3-USPION probes and anti-AFP-USPION probes could specifically enter into the HepG2 cell by combining with the GPC3 receptors or AFP receptors, whereas the HepG2 cell sample incubated with USPIONs showed no or few nanoparticles in the cytoplasm.
The synthesized probes using one-pot method can be used for in vitro experimental study and have potential clinical application in MR imaging for detection of hepatocellular carcinomas.
采用一锅法制备用于检测肝细胞癌（HCC）的特异性磁共振（MR）探针。
通过1-（3-二甲基氨基丙基）-3-乙基碳二亚胺盐酸盐/ N-羟基琥珀酰亚胺（EDC / NHS）介导的反应，将羧化葡聚糖包被的纳米颗粒与抗甲胎蛋白（抗AFP）或抗磷脂酰肌醇蛋白聚糖3（抗GPC3）抗体偶联，以合成探针。通过透射电子显微镜（TEM）和动态光散射测定探针的物理和化学性质，并使用1.5T临床MR扫描仪将弛豫率与未结合的超小超顺磁性氧化铁纳米颗粒（USPIONs）进行比较。用紫外可见分光光度计测量抗体与纳米颗粒的结合效率。此外，将探针与可靶向细胞在体外孵育。
采用一锅法合成了超顺磁性MR探针（抗GPC3-USPION探针和抗AFP-USPION探针）。它们的平均流体动力学直径为47 nm，尺寸分布稍宽。羧化葡聚糖包被的超小超顺磁性氧化铁（USPIO）与抗GPC3或抗AFP抗体的偶联效率分别为15.9％和88.8％。每个USPIO纳米颗粒可以结合3个GPC3抗体或12个AFP抗体。统计分析表明，将USPIO-AFP或USPIO-GPC3与USPIO比较时，在缩短T1和T2值方面无统计学意义（P> 0.05）。TEM图像分析显示，抗GPC3-USPION探针和抗AFP-USPION探针可通过与GPC3受体或AFP受体结合而特异性进入HepG2细胞，而与USPIONs孵育的HepG2细胞样品在细胞质中未显示或仅显示少量纳米颗粒。
采用一锅法合成的探针可用于体外实验研究，在肝细胞癌的MR成像检测中具有潜在的临床应用价值。
