Jummah Nurul, Satrialdi Satrialdi, Artarini Aluicia Anita, Anindyajati Anindyajati, Mudhakir Diky
Department of Pharmaceutics, School of Pharmacy, Institut Teknologi Bandung (ITB), Bandung 40132, Indonesia.
Department of Pharmacy, Faculty of Mathematics and Natural Science, Universitas Islam Makassar, Makassar 90245, Indonesia.
Adv Pharm Bull. 2024 Oct;14(3):613-622. doi: 10.34172/apb.2024.050. Epub 2024 Jun 22.
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.
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.
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.
NLC formulation could deliver genetic material to the nucleus and could be considered a gene therapy candidate for spermatogenesis.
本研究使用增强型绿色荧光蛋白（EGFP）-C1质粒模型评估纳米结构脂质载体（NLC）递送系统能否安全、准确地将核酸递送至细胞核。
采用乳化法制备NLC，并配备1,2-二油酰基-3-三甲基铵丙烷（DOTAP）用于阳离子脂质介导的转染，DOTAP与核酸发生静电相互作用。通过动态光散射（DLS）评估NLC的属性，包括粒径、多分散指数和zeta电位。使用透射电子显微镜分析形态结构。通过直接法评估包封率。使用共聚焦显微镜观察pEGFP-C1-NLC的细胞摄取机制以及pEGFP-C1穿透TM4细胞细胞核以表达EGFP的能力。
pEGFP-C1-NLC的粒径范围为56 - 88 nm，粒子电荷范围为-6.0至+1.3 mV。多分散指数<0.5表明粒径均匀性良好，pEGFP-C1在NLC中的包封率为92.06±2.295%。如NLC成功将pEGFP递送至细胞后EGFP表达所示，NLC制剂主要通过小窝介导的内吞作用内化。
NLC制剂可将遗传物质递送至细胞核，可被视为精子发生的基因治疗候选物。
