PEG shielded polymeric double-layered micelles for gene delivery
PEG shielded polymeric double-layered micelles for gene delivery
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DOI：
10.1016/j.jconrel.2004.11.005
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发表时间：
2005-02-16
影响因子：
10.8
通讯作者：
van Nostrum, CF
中科院分区：
文献类型：
--
作者：
Funhoff, AM;Monge, S;van Nostrum, CF
A-B和B-C嵌段共聚物的组合用于将DNA封装在pEG包被的颗粒内，其中A是用于DNA结合和缩合的阳离子嵌段（聚（甲基丙烯酸二甲氨基乙酯），pDMAEMA），B是疏水性嵌段（聚（甲基丙烯酸丁酯），pBMA），C是聚乙二醇（pEG）嵌段。 AB和BC嵌段共聚物是通过过渡金属介导的自由基聚合合成的。 AB嵌段共聚物具有固定的pBMA分子量3800g/mol和变化的pDMAEMA分子量（从22至65kg/mol），BC嵌段共聚物具有固定的组成（pBMA 9000g/mol；pEG 2000g/mol）。通过去污剂透析方法制备含有质粒DNA的颗粒。通过这种方法，形成了约 120 个边缘的颗粒（通过动态光散射 (DLS) 测定），具有接近中性的电荷，与 DMAEMA 块尺寸无关。 DLS 测量和凝胶电泳表明，该颗粒在 37℃ 的细胞培养基中非常稳定，并且能抵抗聚-L-天冬氨酸的阴离子交换。如果孵育 1 或 4 小时，颗粒能够转染 COS-7 和 OVCAR-3 细胞，且毒性较小；孵育24小时导致毒性增加。本文表明，使用 pDMAEMA-b-pBMA 和 pBMA-b-pEG 通过方便的去污剂透析方法可以获得带有接近中性电荷的小聚合复合物。这些颗粒对于体内实验可能很有趣，其中带有高正电荷的颗粒与血液成分具有不利的相互作用。 (C) 2004 Elsevier B.V. 保留所有权利。
A combination of A-B and B-C block copolymers was used to encapsulate DNA inside pEG coated particles, where A is a cationic block (poly(dimethylaminoethyl methacrylate), pDMAEMA) for DNA binding and condensation, B is a hydrophobic block (poly(butylmethacrylate), pBMA) and C is a polyethylene glycol (pEG) block. The AB and BC block copolymers were synthesized by transition metal mediated radical polymerization. The AB block copolymer had a fixed pBMA molecular weight of 3800 g/mol and a varying pDMAEMA molecular weight (from 22 to 65 kg/mol), the BC block copolymer had a fixed composition (pBMA 9000 g/mol; pEG 2000 g/mol). Plasmid DNA containing particles were made via a detergent dialysis method. By this method, particles of approximately 120 rim, as determined by dynamic light scattering (DLS), with a near neutral charge were formed, independent of the DMAEMA block size. DLS measurements and gel electrophoresis indicated that the particles were very stable in cell culture medium at 37 degreesC and resistant to anionic exchange by poly-L-aspartic acid. The particles were able to transfect COS-7 and OVCAR-3 cells with minor toxicity if incubated for I or 4 h; incubation for 24 h resulted in an increased toxicity. This paper shows that small polyplexes with near neutral charge can be obtained via a convenient detergent dialysis method using pDMAEMA-b-pBMA and pBMA-b-pEG. These particles may be interesting for in vivo experiments where particles with high positive charges have adverse interactions with blood components. (C) 2004 Elsevier B.V. All rights reserved.