Nguyen Hien V, Campbell Katrin, Painter Gavin F, Young Sarah L, Walker Greg F
School of Pharmacy, University of Otago, Dunedin 9016, New Zealand.
Department of Pathology, University of Otago, Dunedin 9016, New Zealand.
Pharmaceutics. 2020 Nov 27;12(12):1150. doi: 10.3390/pharmaceutics12121150.
The aim of this study is to prepare and characterize an amino-dextran nanoparticle (aDNP) platform and investigate two loading strategies for unmethylated cytosine-phosphate-guanine (CpG) oligonucleotide. aDNP was prepared by desolvation of amino-dextran followed by the chemical crosslinking of amino groups. Size, surface charge, and surface morphology of aDNP was determined by dynamic light scattering and transmission electron microscopy. CpG was either loaded onto aDNP by adsorption (CpG-adsorbed-aDNP) or conjugated to aDNP (CpG-conjugated-aDNP). In vitro cytokine production by bone marrow-derived dendritic cells (BMDCs) was measured by flow cytometry. aDNPs size and zeta potential could be controlled to produce uniform particles in the size range of 50 to 300 nm, surface charge of -16.5 to +14 mV, and were spherical in shape. Formulation control parameters investigated included the anti-solvent, water-to-anti-solvent ratio, level of amine functionality of dextran, and the molar ratio of glutaraldehyde to amine. aDNP could be lyophilized without additional cryoprotectant. Unloaded cationic aDNP (+13 mV) showed acceptable in vitro hemolysis. Unloaded and CpG-loaded aDNPs showed no cytotoxicity on BMDCs. CpG-loaded nanoparticles stimulated cytokine production by BMDCs, the level of cytokine production was higher for CpG-conjugated-aDNP compared to CpG-absorbed-aDNP. aDNP is a promising new drug delivery platform as its offers versatility in loading and tuning of particle properties.
本研究的目的是制备并表征一种氨基葡聚糖纳米颗粒（aDNP）平台，并研究未甲基化胞嘧啶-磷酸-鸟嘌呤（CpG）寡核苷酸的两种负载策略。通过氨基葡聚糖的去溶剂化，随后对氨基进行化学交联来制备aDNP。通过动态光散射和透射电子显微镜测定aDNP的尺寸、表面电荷和表面形态。CpG要么通过吸附负载到aDNP上（CpG吸附-aDNP），要么与aDNP共轭（CpG共轭-aDNP）。通过流式细胞术测量骨髓来源的树突状细胞（BMDCs）的体外细胞因子产生。aDNP的尺寸和zeta电位可以控制，以产生尺寸范围为50至300nm、表面电荷为-16.5至+14mV的均匀颗粒，且形状为球形。研究的制剂控制参数包括反溶剂、水与反溶剂的比例、葡聚糖的胺官能团水平以及戊二醛与胺的摩尔比。aDNP可以在不添加额外冷冻保护剂的情况下冻干。未负载的阳离子aDNP（+13mV）在体外显示出可接受的溶血率。未负载和负载CpG的aDNP对BMDCs均无细胞毒性。负载CpG的纳米颗粒刺激BMDCs产生细胞因子，与CpG吸附-aDNP相比，CpG共轭-aDNP的细胞因子产生水平更高。aDNP是一种有前途的新型药物递送平台，因为它在负载和调节颗粒性质方面具有多功能性。
