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目的制备一种以凝溶胶蛋白(GSN)为靶点的包裹液态氟碳的纳米级PLGA高分子造影剂(GSN-PLGA-PFP),并评价其体外热致相变、靶向及显影能力。方法采用双乳化法及碳二亚胺法制备靶向相变型高分子超声造影剂。检测此造影剂的一般性质;显微镜加热板法进行热致相变;激光共聚焦显微镜体外观察GSN-PLGA-PFP被小鼠腹水型肝癌高淋巴道转移Hca-F细胞摄取的情况,初步评价其寻靶能力;体外低强度聚焦超声(LIFU)仪辐照后观察其超声显影效果。结果 GSN-PLGA-PFP的平均粒径为(328.59±3.82)nm,造影剂表面平均Zeta电位为(-10.36±1.34)mV,造影剂表面GSN单抗连接率高达98.31%,且于43.5℃时纳米粒开始发生相变;体外摄取实验显示Hca-F细胞可较多地摄取GSNPLGA-PFP,体外行LIFU辐照后可观察到显影效果明显增强。结论自制携Gelsolin单抗包载PFP的PLGA造影剂热致相变效果明显,可与Hca-F细胞特异度结合且体外显影效果较好。
Objective To prepare a nano-scale PLGA polymeric contrast agent (GSN-PLGA-PFP) encapsulating liquid fluorocarbon targeting gelsolin (GSN) and evaluate its in vitro pyrogenic phase-changing, targeting and developing ability. Methods Double-emulsion method and carbodiimide method were used to prepare targeted phase-change polymer ultrasound contrast agent. The general properties of this contrast agent were examined. The phase change of the plate was observed by the heating method of microscope. The uptake of GSN-PLGA-PFP by Hca-F cells in mouse ascites hepatocarcinoma with high lymphatic metastasis was observed by laser scanning confocal microscopy. Targeting ability; low intensity focused ultrasound (LIFU) instrument after irradiation to observe the effect of ultrasound imaging. Results The mean particle size of GSN-PLGA-PFP was (328.59 ± 3.82) nm, the average Zeta potential of the contrast agent was (-10.36 ± 1.34) mV, and the conjugation rate of GSN McAb was 98.31% In vitro experiments showed that Hca-F cells could ingest more GSNPLGA-PFP, and the development effect of LFPU in vitro could be observed. CONCLUSION: The PLGA contrast agent loaded with Gelsolin mAb can significantly induce thermal induced phase transition, and can specifically bind to Hca-F cells with better imaging in vitro.