Hangjie Wang, Wei Li, Junyan Chen, Rong Chen, Yuwei Qi, Linshuang Shen, Kaidi Chen, Lewei Dai, Yuxin Sheng, An Wang, Hong Wang, Chujian Chen, Xiao Cheng, Mancang Gu
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引用次数: 0
Abstract
Context: Lipid nanoparticles (LNPs) are the primary non-viral vectors for siRNA delivery. However, synthetic lipids face issues, such as low lysosomal escape efficiency and high cost.
Objective: This study aimed to use three natural lipids to construct LNPs, optimize their preparation and freeze-drying processes, and evaluate their siRNA delivery efficiency in vitro.
Materials and methods: Coix seed lipid [Coix lacryma-jobi L. var. mayuen (Roman.) Stapf (Poaceae), CSL], Brucea javanica seed lipid [Brucea javanica (L.) Merr. (Simaroubaceae), BJL], and Soybean oil [Glycine max (L.) Merr. (Fabaceae), SO] were used to construct LNPs. The Z-average size, zeta potential, Polymer Dispersity Index, and N/P ratio of the LNPs were characterized. Transmission electron microscope was used for morphology observation and the MTS assay for cytotoxicity. Confocal laser scanning microscope assessed cell uptake, lysosomal escape, and co-localization of lipid droplets. The efficiency of siRNA knockdown was evaluated in three cells using qPCR and Western blot. The freeze-drying processes were optimized.
Results: The optimal LNPs exhibited a size of 160-180 nm, zeta of 44-50 mV, and PDI of <0.2. At 200 μg/mL, the LNPs did not affect cell viability. CSL-LNPs, BJL-LNPs, and SO-LNPs reduced KRASG12D mRNA levels in AsPC-1 cells by 67.87 ± 3.89, 47.18 ± 7.65, and 42.52 ± 8.90%, respectively. Freeze-dried LNPs retained their basic physical properties and the three LNPs reducing KRASG12D mRNA levels by 58.47 ± 4.00, 51.83 ± 4.57, and 38.00 ± 4.89%, respectively.
Discussion and conclusion: Natural lipids are promising components for LNPs construction, offering new avenues for siRNA delivery in gene therapy.
期刊介绍：
Pharmaceutical Biology will publish manuscripts describing the discovery, methods for discovery, description, analysis characterization, and production/isolation (including sources and surveys) of biologically-active chemicals or other substances, drugs, pharmaceutical products, or preparations utilized in systems of traditional medicine.
Topics may generally encompass any facet of natural product research related to pharmaceutical biology. Papers dealing with agents or topics related to natural product drugs are also appropriate (e.g., semi-synthetic derivatives). Manuscripts will be published as reviews, perspectives, regular research articles, and short communications. The primary criteria for acceptance and publication are scientific rigor and potential to advance the field.
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