Department of Pharmaceutical Sciences, School of Pharmacy, Northeastern University, Boston, Massachusetts 02115, United States.
Biomacromolecules. 2012 Apr 9;13(4):1074-85. doi: 10.1021/bm2017993. Epub 2012 Mar 19.
The main objective of this study was to evaluate macrophage-targeted alginate nanoparticles as a noncondensing gene delivery system for potential anti-inflammatory therapy. An external gelation method was employed to form plasmid DNA-encapsulated alginate nanoparticles. The nanoparticle surface was modified with a peptide sequence containing tuftsin (TKPR), and transfection efficiency was determined in J774A.1 macrophages. The effect of transfected mIL-10 in blocking expression of tumor necrosis factor-alpha (TNF-α) was evaluated in lipopolysaccharide (LPS)-stimulated cells. Scrambled peptide- and tuftsin-modified cross-linked alginate nanoparticles efficiently encapsulated plasmid DNA and protected against DNase I degradation. The transgene expression efficiencies, measured using GFP and mIL-10 expressing plasmid DNA, were highest with tuftsin-modified nanoparticles. Levels of TNF-α were significantly lower (p < 0.0001) in LPS-stimulated cells that were transfected with mIL-10 using alginate nanoparticles. The results of the study show that noncondensing alginate nanoparticles can efficiently deliver plasmid DNA, leading to sustained in vitro gene expression in macrophages.
本研究的主要目的是评估巨噬细胞靶向海藻酸钠纳米粒作为一种非凝结基因传递系统，用于潜在的抗炎治疗。采用外部凝胶化方法形成包封质粒 DNA 的海藻酸钠纳米粒。纳米粒表面用含有 tuftsin（TKPR）的肽序列进行修饰，并在 J774A.1 巨噬细胞中测定转染效率。用转染的 mIL-10 阻断脂多糖（LPS）刺激细胞中肿瘤坏死因子-α（TNF-α）表达的效果进行了评估。用 scrambled 肽和 tuftsin 修饰的交联海藻酸钠纳米粒有效地包封了质粒 DNA 并防止了 DNAse I 降解。使用 GFP 和 mIL-10 表达质粒 DNA 测量的转基因表达效率，tuftsin 修饰的纳米粒最高。用海藻酸钠纳米粒转染 mIL-10 的 LPS 刺激细胞中 TNF-α 的水平显著降低（p < 0.0001）。研究结果表明，非凝结海藻酸钠纳米粒可以有效地传递质粒 DNA，导致巨噬细胞中持续的体外基因表达。
