Development of a non-viral gene vector for enhancing gene transfection efficiency

• Published in: Journal of drug delivery science and technology 2022-09, Vol.75, p.103669, Article 103669
• Main Authors: Li, Yue, Yu, Ting, Han, Long-zhe, Jin, Li-li, Jin, Yong, Quan, Ji-shan
• Format: Article
• Language: English
• Subjects: Cytotoxicity; Endocytosis; Lipopolyplexes; Transfection efficiency
• ISSN: 1773-2247
• DOI: 10.1016/j.jddst.2022.103669

Construction of a safe and high transfection efficiency non-viral gene delivery vector to improve the effectiveness of gene delivery is vital for cancer treatment. Herein, we used 1, 2-Dioleoyl-3-Trimethylammonium-Propane (DOTAP), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and cholesterol to prepare cationic liposomes in combination with poly(aspartate-graft-PEI423) (PAE) and plasmid DNA to prepare Liposomes-poly(aspartate-graft-PEI423)/DNA (LP-PAE/DNA) to improve gene transfection efficiency. LP-PAE/DNA particle size was about 220 nm, the zeta potential was about 20 mV and the morphology was spherical. The LP-PAE/DNA lipopolyplexes showed higher transfection efficiency at a weight ratio of 10:1:1 in HeLa, A549 and SPC-A1 cell line. LP-PAE/DNA lipopolyplexes toxicity in different cell lines and the main endocytic pathways through which LP and LP-PAE mediate DNA into cells was examined. In addition, the results of hemolysis test, red blood cell morphology observation and blood routine experiments showed that LP-PAE/DNA would not cause systemic hematological toxicity. The histological analysis including hematoxylin-eosin (HE) staining and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay confirmed lipopolyplexes/liposomes mediate the entry of therapeutic genes into the body not causing damage to normal organs and enhancing tumor tissue apoptosis relative to more traditional approaches. Therefore, it could be hoped the feasibility of LP-PAE as an efficient and promising platform for gene delivery. [Display omitted] •Lipopolyplexes formulation (LP:PAE:DNA) is in a weight ratio of 10:1:1 with favorable genetic drug delivery characteristics.•Lipopolyplexes show enhanced transfection effect than liposomes in HeLa, A549 and SPC-A1 cell line.•Lipopolyplexes show good biocompatibility and low toxicity.•Cellular uptake pathway for LP-PAE/DNA is clathrin-mediated endocytosis and caveolae-mediated endocytosis.