Del Prado Anselmo, Civantos Ana, Martínez-Campos Enrique, Levkin Pavel A, Reinecke Helmut, Gallardo Alberto, Elvira Carlos
Instituto de Ciencia y Tecnología de Polímeros, CSIC, Juan de la Cierva 3, 28006 Madrid, Spain.
Departamento de Química Orgánica, Facultad de Ciencias, Universidad Autónoma de Madrid, 28049 Madrid, Spain.
Polymers (Basel). 2020 Nov 17;12(11):2724. doi: 10.3390/polym12112724.
Non-viral vectors are a safety tool for gene therapy to deliver therapeutic genes. Among the different non-viral vectors, polyvinylpyrrolidone (PVP), a well-known hydrosoluble, neutral, and non-toxic polymer, satisfies the requirements and becomes a suitable candidate for gene delivery. In this study, we describe the preparation of polyvinylpyrrolidones decorated with pyrrolidine, piperidine, and piperazine groups, and evaluate them in vitro as non-viral gene carriers. The properties of these new systems are compared with those of hyperbranched polyethyleneimine (PEI) used as a positive control. Their ability to complex DNA at different N/P molar ratios, from 1:1 up to 10:1, was studied through agarose gel electrophoresis and dynamic light scattering. The resulting complexes (polyplexes) were characterized and evaluated in vitro with murine fibroblast (Swiss 3T3) as non-viral gene carriers, using luciferase as the reporter gene and a calcein cytocompatibility assay. All the copolymers condensed DNA to a particle average size between 100-400 nm when used at N/P ratios of 4:1 or higher. The copolymers with piperidine groups showed higher transfection efficiency than the pyrrolidine and piperazine modified copolymers, and even higher than the positive control of PEI at N/P ratios of 4:1 or higher. All the synthesized polyplexes from an aminated PVP displayed a general tendency of high cytocompatibility (75-95%) in comparison with the positive control PEI (55%).
非病毒载体是用于基因治疗以递送治疗性基因的一种安全工具。在不同的非病毒载体中，聚乙烯吡咯烷酮（PVP），一种众所周知的水溶性、中性且无毒的聚合物，满足相关要求，成为基因递送的合适候选者。在本研究中，我们描述了用吡咯烷、哌啶和哌嗪基团修饰的聚乙烯吡咯烷酮的制备，并在体外将它们评估为非病毒基因载体。将这些新体系的性质与用作阳性对照的超支化聚乙烯亚胺（PEI）的性质进行比较。通过琼脂糖凝胶电泳和动态光散射研究了它们在1:1至10:1的不同N/P摩尔比下与DNA复合的能力。以荧光素酶作为报告基因并采用钙黄绿素细胞相容性测定法，将所得复合物（多聚体）作为非病毒基因载体，用小鼠成纤维细胞（瑞士3T3）在体外进行表征和评估。当以4:1或更高的N/P比使用时，所有共聚物都将DNA浓缩成平均粒径在100 - 400 nm之间的颗粒。含哌啶基团的共聚物显示出比含吡咯烷和哌嗪修饰的共聚物更高的转染效率，并且在4:1或更高的N/P比下甚至高于PEI阳性对照。与阳性对照PEI（55%）相比，所有由胺化PVP合成的多聚体都表现出高细胞相容性（75 - 95%）的总体趋势。
