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空的或负载基因的脂多糖 - 胺纳米聚合物囊泡的稳定性和毒性

Stability and toxicity of empty or gene-loaded lipopolysaccharide-amine nanopolymersomes.

作者信息

Wang Qinmei, Chen Ying, Wang Lichun, Zhang Xinchun, Huang Hongzhang, Teng Wei

机构信息

Key Laboratory on Assisted Circulation, Ministry of Health, Cardiovascular Division, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, People's Republic of China.

Hospital of Stomatology, Institute of Stomatological Research, Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou, People's Republic of China.

出版信息

Int J Nanomedicine. 2015 Jan 13;10:597-608. doi: 10.2147/IJN.S74156. eCollection 2015.

Abstract

Successful in vivo gene delivery mediated by nonviral vectors requires efficient extracellular and intracellular gene delivery, but few studies have given attention to the former. That is why numerous gene delivery systems have succeeded in vitro, while the expected clinical success has not come about. To realize efficient extracellular gene delivery, the stability of vectors and/or their complexes with genes in body fluids is first required, which prevents loaded genes from premature unloading and degradation. Furthermore, the storage stability of vectors under common conditions is important for their widespread applications. Lipopolysaccharide-amine nanopolymersomes (NPs), a gene vector developed by our group recently, have higher than 95% in vitro transfection efficiency in mesenchymal stem cells when delivering pEGFP, and induce significant angiogenesis in zebrafish when delivering plasmid encoding vascular endothelial growth factor deoxyribonucleic acid (pVEGF). To reveal their extracellular delivery ability and storage stability, in this study their stability in various simulant physiological environments and storage conditions was systematically studied by monitoring their changes in disassembly, size, zeta potential, and transfection efficiency. Additionally, damage to the mitochondria of mesenchymal stem cells was evaluated. Results show that NPs and plasmid deoxyribonucleic acid (pDNA)-loaded NPs (pNPs) have acceptable stability against dilution, anions, salts, pH, enzyme, and serum, presumably assuring their efficient extracellular delivery in vivo. Moreover, both the lyophilized NPs at room temperature and NP/pNP solution at 4°C have high storage stability, and pNPs show low damage to the mitochondria. The acceptable stability of NPs combined with compatibility and efficient gene transfection highlight their huge potential in the clinic as a gene delivery vector.

摘要

非病毒载体介导的体内基因递送成功需要高效的细胞外和细胞内基因递送,但很少有研究关注前者。这就是为什么众多基因递送系统在体外取得成功,而预期的临床成功却尚未实现的原因。为了实现高效的细胞外基因递送,首先需要载体及其与基因在体液中的复合物具有稳定性,这可防止负载的基因过早卸载和降解。此外,载体在常见条件下的储存稳定性对于其广泛应用也很重要。脂多糖 - 胺纳米聚合物囊泡(NPs)是我们小组最近开发的一种基因载体,在递送pEGFP时,在间充质干细胞中的体外转染效率高于95%,在递送编码血管内皮生长因子脱氧核糖核酸的质粒(pVEGF)时,可在斑马鱼中诱导显著的血管生成。为了揭示它们的细胞外递送能力和储存稳定性,在本研究中,通过监测它们在解体、大小、zeta电位和转染效率方面的变化,系统地研究了它们在各种模拟生理环境和储存条件下的稳定性。此外,还评估了对间充质干细胞线粒体的损伤。结果表明,NPs和负载质粒脱氧核糖核酸(pDNA)的NPs(pNPs)在抗稀释、阴离子、盐、pH、酶和血清方面具有可接受的稳定性,这大概确保了它们在体内的高效细胞外递送。此外,室温下的冻干NPs和4°C下的NP/pNP溶液都具有高储存稳定性,并且pNPs对线粒体的损伤较小。NPs可接受的稳定性与兼容性和高效基因转染相结合,突出了它们作为基因递送载体在临床上的巨大潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76c0/4298338/218b373d7018/ijn-10-597Fig1.jpg

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