Department of Biomedical Engineering, Yale University, New Haven, CT 06511, USA.
J Control Release. 2012 Nov 28;164(1):41-8. doi: 10.1016/j.jconrel.2012.09.020. Epub 2012 Oct 5.
The use of biodegradable polymers provides a potentially safe and effective alternative to viral and liposomal vectors for the delivery of plasmid DNA to cells for gene therapy applications. In this work we describe the formulation of a novel nanoparticle (NP) system containing a blend of poly(lactic-co-glycolic acid) and a representative poly(beta-amino) ester (PLGA and PBAE respectively) for use as gene delivery vehicles. Particles of different weight/weight (wt/wt) ratios of the two polymers were characterized for size, morphology, plasmid DNA (pDNA) loading and surface charge. NPs containing PBAE were more effective at cellular internalization and transfection (COS-7 and CFBE41o-) than NPs lacking the PBAE polymer. However, along with these delivery benefits, PBAE exhibited cytotoxic effects that presented an engineering challenge. Surface coating of these blended particles with the cell-penetrating peptides (CPPs) mTAT, bPrPp and MPG via a PEGylated phospholipid linker (DSPE-PEG2000) resulted in NPs that reduced surface charge and cellular toxicity to levels comparable with NPs formulated with only PLGA. Additionally, these coated nanoparticles showed an improvement in pDNA loading, intracellular uptake and transfection efficiency, when compared to NPs lacking the surface coating. Although all particles with a CPP coating outperformed unmodified NPs, respectively, bPrPp and MPG coating resulted in 3 and 4.5× more pDNA loading than unmodified particles and approximately an order of magnitude improvement on transfection efficiency in CFBE41o- cells. These results demonstrate that surface-modified PBAE containing NPs are a highly effective and minimally toxic platform for pDNA delivery.
可生物降解聚合物的使用为将质粒 DNA 递送到细胞中用于基因治疗应用的病毒和脂质体载体提供了一种潜在的安全有效的替代方法。在这项工作中，我们描述了一种新型纳米颗粒 (NP) 系统的配方，该系统包含聚（乳酸-共-乙醇酸）和代表性的聚（β-氨基）酯（PLGA 和 PBAE 分别）的混合物，用作基因传递载体。对不同重量/重量 (wt/wt) 比的两种聚合物的粒子进行了大小、形态、质粒 DNA (pDNA) 负载和表面电荷的表征。含有 PBAE 的 NPs 比不含 PBAE 聚合物的 NPs 更有效地进行细胞内化和转染（COS-7 和 CFBE41o-）。然而，除了这些传递益处之外，PBAE 还表现出细胞毒性作用，这给工程带来了挑战。通过 PEG 化磷脂链接（DSPE-PEG2000）将这些混合粒子的表面用穿透肽 (CPP) mTAT、bPrPp 和 MPG 进行涂层，得到的 NPs 降低了表面电荷和细胞毒性，使其与仅用 PLGA 配制的 NPs 相当。此外，与没有表面涂层的 NPs 相比，这些涂覆的纳米颗粒在 pDNA 负载、细胞内摄取和转染效率方面表现出改善。尽管所有带有 CPP 涂层的粒子的性能均优于未修饰的 NPs，但 bPrPp 和 MPG 涂层分别使 pDNA 负载增加了 3 倍和 4.5 倍，并且在 CFBE41o-细胞中的转染效率提高了约一个数量级。这些结果表明，表面修饰的含 PBAE 的 NPs 是一种高效且毒性最小的 pDNA 传递平台。
