NLC 将 EGFP 质粒输送到 TM4 细胞核,实现靶向基因治疗。

IF 3.1 Q2 PHARMACOLOGY & PHARMACY Advanced pharmaceutical bulletin Pub Date : 2024-10-01 Epub Date: 2024-06-22 DOI:10.34172/apb.2024.050
Nurul Jummah, Satrialdi Satrialdi, Aluicia Anita Artarini, Anindyajati Anindyajati, Diky Mudhakir
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引用次数: 0

摘要

目的:本研究以增强型绿色荧光蛋白(EGFP)-C1质粒为模型,评估了纳米结构脂质载体(NLC)递送系统能否安全、准确地将核酸递送至细胞核:方法:采用乳化法配制了NLC,并在阳离子脂质介导的转染中加入了1,2-二油酰-3-三甲基铵丙烷(DOTAP),DOTAP能与核酸发生静电相互作用。通过动态光散射(DLS)评估了 NLC 的属性,包括尺寸、多分散指数和 zeta 电位。透射电子显微镜分析了其形态结构。采用直接法评估了包埋效率。使用共聚焦显微镜观察了 pEGFP-C1-NLC 的细胞摄取机制以及 pEGFP-C1 穿透 TM4 细胞核表达 EGFP 的能力。结论:NLC 配方可将遗传物质输送到细胞内:NLC制剂能将遗传物质输送到细胞核,可作为精子发生基因治疗的候选药物。
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NLC Delivery of EGFP Plasmid to TM4 Cell Nuclei for Targeted Gene Therapy.

Purpose: This study evaluated whether a nanostructured lipid carrier (NLC) delivery system could safely and accurately deliver nucleic acids to the cell nucleus using the enhanced green fluorescent protein (EGFP)-C1 plasmid model.

Methods: The NLC was formulated using the emulsification method and equipped for cationic lipid-mediated transfection with 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), which interacts electrostatically with nucleic acid. The NLC attributes, including size, polydispersity index, and zeta potential, were assessed by dynamic light scattering (DLS). The morphological structure was analyzed using transmission electron microscopy. Entrapment efficiency was evaluated by a direct method. Cellular uptake mechanisms of pEGFP-C1-NLC and the ability of pEGFP-C1 to penetrate the nucleus of TM4 cells to express EGFP were observed using confocal microscopy.

Results: pEGFP-C1-NLC exhibited particle sizes in the range 56-88 nm with a particle charge range of -6.0 to+1.3 mV. The polydispersity index<0.5 showed good size uniformity, and entrapment efficiency of pEGFP-C1in the NLC was 92.06±2.295%. The NLC formulation was internalized predominantly via caveolae-mediated endocytosis, as indicated by EGFP expression following successful delivery of pEGFP by the NLC into the cells.

Conclusion: NLC formulation could deliver genetic material to the nucleus and could be considered a gene therapy candidate for spermatogenesis.

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来源期刊
Advanced pharmaceutical bulletin
Advanced pharmaceutical bulletin PHARMACOLOGY & PHARMACY-
CiteScore
6.80
自引率
2.80%
发文量
51
审稿时长
12 weeks
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