Clima Lilia, Craciun Bogdan Florin, Gavril Gabriela, Pinteala Mariana
"Petru Poni" Institute of Macromolecular Chemistry, Romanian Academy, Centre of Advanced Research in Bionanoconjugates and Biopolymers, Grigore Ghica Voda Alley, 41 A, 700487 Iasi, Romania.
Polymers (Basel). 2019 Aug 6;11(8):1313. doi: 10.3390/polym11081313.
Polyethylene glycol (PEG) functionalization of non-viral vectors represents a powerful tool through the formation of an overall surface charge shielding ability, which is fundamental for efficient nucleic acid delivery systems. The degree of non-viral vector PEGylation and the molecular weight of utilized PEG is crucial since the excessive use of PEG units may lead to a considerable reduction of the DNA-binding capacity and, subsequently, in a reduction of in vitro transfection efficiency. Herein, we report a detailed study on a series of dynamic combinatorial frameworks (DCFs) containing PEGylated squalene, poly-(ethyleneglycol)-bis(3-aminopropyl) of different lengths, and branched low molecular weight polyethylenimine components, reversibly connected in hyperbranched structures, as efficient dynamic non-viral vectors. The obtained frameworks were capable of forming distinct supramolecular amphiphilic architectures, shown by transmission electron microscopy (TEM) and dynamic light scattering (DLS), with sizes and stability depending on the length of PEG units. The interaction of PEGylated DCFs with nucleic acids was investigated by agarose gel retardation assay and atomic force microscopy (AFM), while their transfection efficiency (using pCS2+MT-Luc DNA as a reporter gene) and cytotoxicity were evaluated in HeLa cells. In addition, the data on the influence of the poly-(ethyleneglycol)-bis(3-aminopropyl) length in composition of designed frameworks over transfection efficiency and tolerance in human cells were analyzed and compared.
非病毒载体的聚乙二醇（PEG）功能化是一种强大的工具，可通过形成整体表面电荷屏蔽能力来实现，这对于高效核酸递送系统至关重要。非病毒载体的PEG化程度和所用PEG的分子量至关重要，因为过度使用PEG单元可能会导致DNA结合能力大幅下降，进而降低体外转染效率。在此，我们报告了一项关于一系列动态组合框架（DCF）的详细研究，这些框架包含不同长度的聚乙二醇化角鲨烯、聚（乙二醇）-双（3-氨丙基）以及支链低分子量聚乙烯亚胺成分，它们以超支化结构可逆连接，作为高效的动态非病毒载体。通过透射电子显微镜（TEM）和动态光散射（DLS）表明，所获得的框架能够形成独特的超分子两亲结构，其尺寸和稳定性取决于PEG单元的长度。通过琼脂糖凝胶阻滞试验和原子力显微镜（AFM）研究了聚乙二醇化DCF与核酸的相互作用，同时在HeLa细胞中评估了它们的转染效率（使用pCS2 + MT-Luc DNA作为报告基因）和细胞毒性。此外，还分析并比较了设计框架组成中聚（乙二醇）-双（3-氨丙基）长度对转染效率和人体细胞耐受性影响的数据。
