In Vitro Gene Transfection with Surface-Modified Gelatin Nanoparticles

Klaus Zwiorek, Julia Kloeckner, E. Wagner, C. Coester
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引用次数: 2

Abstract

With the development of the two step desolvation method for the preparation of gelatin nanoparticles, it is possible to reproducibly generate homogeneous colloidal spheres. After the establishment of a surface modification to achieve stable nucleotide loading onto the particles, the goal of this study was to test this new biodegradable and simple producible non-viral gene delivery system in vitro. Two different types of gelatin nanoparticles, referring to size and zeta (ζ) potential were investigated. Therefore, we varied conditions as for example the loaded DNA amount and the conjugation media to find a preferable setup. All preparations were tested on B16F10 murine melanoma cells using pCMVLuc as reporter gene. To evaluate our results, we used commonly known, but non-biodegradable, polyethylenimine (PEI) polyplexes as "gold standard" for in vitro transfection. Additionally, we performed accompanying cell viability assays and hemolysis studies with the preparations tested to substantiate the thesis of low cell toxicity of gelatin nanoparticles. Different setups resulted in efficient gene delivery. The achieved levels of gene expression were good but lower as with optimized PEI polyplexes. Nevertheless, the already achieved results show gelatin nanoparticles as promising biodegradable alternative to existing non-viral gene delivery systems.
表面修饰明胶纳米颗粒的体外基因转染
随着两步脱溶法制备明胶纳米颗粒的发展,制备均匀的胶体球成为可能。在建立表面修饰以实现稳定的核苷酸装载到颗粒上之后,本研究的目标是在体外测试这种新的可生物降解且简单可生产的非病毒基因传递系统。研究了两种不同类型的明胶纳米颗粒的大小和ζ (ζ)电位。因此,我们改变条件,例如负载的DNA量和结合介质,以找到一个较好的设置。以pCMVLuc为报告基因,在B16F10小鼠黑色素瘤细胞上进行实验。为了评估我们的结果,我们使用了众所周知的,但不可生物降解的聚乙烯亚胺(PEI)多聚物作为体外转染的“金标准”。此外,我们还进行了伴随的细胞活力测定和溶血研究,以证实明胶纳米颗粒的低细胞毒性。不同的设置导致了有效的基因传递。获得的基因表达水平较好,但与优化的PEI多聚体相比较低。然而,已经取得的结果表明,明胶纳米颗粒是现有非病毒基因传递系统的有前途的可生物降解替代品。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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