Department of Pharmacy, School of Pharmacy and Medical Technology, Putian University, Putian, China/Key Laboratory of Pharmaceutical Analysis and Laboratory Medicine in University of Fujian Province, Putian University, Putian, China.
Department of Medicinal Chemistry, School of Pharmacy, Fujian Medical University, Fuzhou, China/Fujian Key Laboratory of Drug Target Discovery and Structural and Functional Research, Fuzhou, China.
Pak J Pharm Sci. 2024 Jul;37(4):903-915.
The present study employed lysine as a modifying agent for chitosan (CS) to synthesise a novel CS derivative (LGCS) intended for siRNA delivery. The successful grafting of lysine to CS was characterized using FT-IR and the introduction of the lysine moiety resulted in improved solubility and buffering capacity of CS. The Zeta potential and size of LGCS/siRNA nanoparticles (NPs) were evaluated using dynamic light scattering (DLS) and the results were verified by transmission electron microscopy (TEM). Evaluation of LGCS's siRNA binding capacity was conducted using a gel retardation assay. The results showed that LGCS could effectively bind to siRNA and form a complex with a hydrated diameter of about 97.2 ± 1.3 nm. Furthermore, cytotoxicity assays conducted on RSC96 cells demonstrated that LGCS exhibited lower toxicity compared to linear polyethyleneimine (PEI) 25k. In vitro, cellular uptake assays also revealed that LGCS displayed excellent transfection efficiency. The results of our study lead us to the conclusion that LGCS holds great promise as a gene delivery vector.
本研究采用赖氨酸作为壳聚糖（CS）的修饰剂，合成了一种新型 CS 衍生物（LGCS），旨在用于 siRNA 的递送。成功地将赖氨酸接枝到 CS 上，通过傅里叶变换红外光谱（FT-IR）进行了表征，并且赖氨酸部分的引入提高了 CS 的溶解度和缓冲能力。通过动态光散射（DLS）评估了 LGCS/siRNA 纳米粒子（NPs）的 Zeta 电位和粒径，并用透射电子显微镜（TEM）进行了验证。通过凝胶阻滞实验评估了 LGCS 的 siRNA 结合能力。结果表明，LGCS 可以有效地与 siRNA 结合，形成一种具有约 97.2 ± 1.3nm 水合直径的复合物。此外，在 RSC96 细胞上进行的细胞毒性试验表明，与线性聚乙烯亚胺（PEI）25k 相比，LGCS 的毒性较低。体外细胞摄取实验也表明，LGCS 具有优异的转染效率。我们的研究结果表明，LGCS 作为一种基因传递载体具有很大的应用潜力。
