Poly(epsilon-lysine) dendrons and nucleic acids complexes for non-viral delivery of bacteriophage DNA into bacterial cells

IF 4.2 Q2 NANOSCIENCE & NANOTECHNOLOGY
Orode UV Aniejurengho, S. Meikle, M. Santin
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引用次数: 0

Abstract

Phage therapy has been advocated as an alternative to antibiotics in the treatment of bacterial infections. However, this approach can be affected by batch-to-batch variability in the harvesting, isolation and storage of specie-specific bacteriophages. The present paper presents the development of synthetic nanostructured carriers that can form complexes with isolated bacteriophage DNA, while ensuring the penetration of the carrier through bacterial wall and membrane. The complexation capability of these poly(epsilon-lysine) dendrons with bacteriophage DNA was tested by mixing the hyperbranched molecules at various charge ratios (132:1 to 0.06:1 of positively charged carriers to negatively charged DNA). These electrostatic complexes were assessed by gel retardation, ethidium bromide displacement assay and transmission electron microscopy. The self-assembly of poly(epsilon-lysine) dendrons with the nucleic acids led to charge ratio-dependent electrostatic complexes. Transfection of both complexed and naked DNA in Proteus mirabilis bacteria demonstrated that the viral DNA maintained its replicative ability with the formation of whole viral particles. Additionally, the dendron’s cytotoxicity was assayed by lactate dehydrogenase (LDH) release and 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl-tetrazolium bromide (MTT) reduction assays. In both assays, 15% cytotoxicity and even lower (6%) necrotic effects were observed by MTT and LDH respectively. Overall, these findings demonstrate that it is possible to complex bacteriophage DNA with poly(epsilon-lysine) dendrons, of three branching generations (Gen3K), and suggests that Gen3K has the potential to be used as a reliable alternative to antibiotics.
多聚赖氨酸树突和核酸复合物,用于将噬菌体DNA非病毒递送到细菌细胞中
噬菌体疗法一直被提倡作为抗生素治疗细菌感染的替代方法。然而,这种方法可能受到收获、分离和储存特定物种噬菌体的批次差异的影响。本文介绍了合成纳米结构载体的发展,该载体可以与分离的噬菌体DNA形成复合物,同时确保载体穿透细菌壁和膜。这些多聚赖氨酸树突与噬菌体DNA的络合能力通过以不同的电荷比(132:1至0.06:1带正电的载体与带负电的DNA)混合这些超支化分子来测试。通过凝胶阻滞、溴化乙锭置换法和透射电镜对这些静电配合物进行了评价。聚赖氨酸树突与核酸的自组装导致电荷比依赖的静电复合物。在奇异变形杆菌中转染复合DNA和裸DNA表明,病毒DNA在形成完整病毒颗粒时保持了复制能力。此外,通过乳酸脱氢酶(LDH)释放和3-(4,5-二甲基噻唑-2-基)2,5-二苯基溴化四唑(MTT)还原测定树突的细胞毒性。在这两项试验中,MTT和LDH分别观察到15%的细胞毒性和更低(6%)的坏死作用。总的来说,这些发现表明,可以将噬菌体DNA与三个分支代(Gen3K)的聚赖氨酸(epsilon-lysine)树突复杂,并表明Gen3K有可能被用作抗生素的可靠替代品。
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来源期刊
CiteScore
6.00
自引率
1.70%
发文量
24
期刊介绍: Proceedings of the Institution of Mechanical Engineers Part N-Journal of Nanomaterials Nanoengineering and Nanosystems is a peer-reviewed scientific journal published since 2004 by SAGE Publications on behalf of the Institution of Mechanical Engineers. The journal focuses on research in the field of nanoengineering, nanoscience and nanotechnology and aims to publish high quality academic papers in this field. In addition, the journal is indexed in several reputable academic databases and abstracting services, including Scopus, Compendex, and CSA's Advanced Polymers Abstracts, Composites Industry Abstracts, and Earthquake Engineering Abstracts.
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