Folate-PEG-folate-graft-polyethylenimine-based gene delivery
Folate-PEG-folate-graft-polyethylenimine-based gene delivery
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DOI：
10.3109/10611860108997923
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发表时间：
2001-01-01
影响因子：
4.5
通讯作者：
Kim, SW
中科院分区：
文献类型：
--
作者：
Benns, JM;Maheshwari, A;Kim, SW
通过将叶酸连接到单官能PEG的两端，然后接枝到PEI上，合成了叶酸-聚乙二醇-叶酸-接枝-聚乙烯亚胺（FPF-g-PEI）。使用比尔定律通过测量363 nm处的UV吸光度来确定接枝率。测定了载体的pH分布、直径和形状。利用荧光素酶报告基因质粒DNA在正常平滑肌细胞（SMC）和CT 26结肠腺癌细胞中优化转染效率。游离叶酸在中性电荷比下抑制FPF-2.3g-PEI的转染。MTT比色法测定PEI与修饰载体的相对毒性。使用ELISA和RT PCR在蛋白质和mRNA水平上测定与这些聚合物载体复合的pmIFN-γ在基因表达方面的治疗潜力。确定FPF-g-PEI具有接枝到每个PEI分子上的2.3个叶酸-PEG-叶酸（FPF）线性聚合物。测得的平均分子量类似于33，500 Mw，pH曲线是内体破坏能力的特征。原子力显微镜（AFM）和动态激光光散射（DLLS）表明FPF-2.3g-PEI和PEI（以2 w/w的比例）有效地浓缩质粒DNA，产生平均直径接近150 nm的斜球形聚合物。FPF-2.3g-PEI在细胞毒性和癌细胞转染效率方面优于PEI上级。平滑肌细胞对叶酸栓系复合物没有特异性，其中PEI/pLuc复合物产生更高的效率。
Folate-polyethylene glycol-folate-grafted-polyethylenimine (FPF-g-PEI), was synthesized by linking folic acid to both ends of a mono-functional PEG and then grafting to PEI. The graft ratio was determined using Beer's law by measuring the UV absorbance at 363 nm. The pH profile, diameter and shape of the carriers were determined. Transfection efficiencies were optimized in normal smooth muscle cells (SMC) and CT 26 colon adenocarcinoma cells using plasmid DNA encoding luciferase reporter gene. Free folic acid was shown to inhibit transfection with FPF-2.3g-PEI at neutral charge ratio. Relative toxicity between PEI and the modified carrier was measured using MTT colorimetric assay. Therapeutic potential of pmIFN-gamma complexed with these polymeric carriers in terms of gene expression was determined at protein and mRNA levels using ELISA and RT PCR. FPF-g-PEI was determined to have 2.3 folate-PEG-folate (FPF) linear polymers grafted to each PEI molecule. The average molecular weight was measured to be similar to 33,500 Mw and the pH profile was characteristic of endosomal disruption capacity. Atomic Force Microscopy (AFM) and Dynamic Laser Light Scattering (DLLS) indicated FPF-2.3g-PEI and PEI (at 2 w/w ratio) efficiently condensed plasmid DNA resulting in oblique spheroid polyplexes with a mean diameter of similar to 150 nm. FPF-2.3g-PEI was superior to PEI in terms of cytotoxicity and transfection efficiency in cancer cells. Smooth muscle cells showed no specificity for folate tethered complexes, where PEI/pLuc complexes yielded higher efficiencies.