Preparation, characterization, and evaluation of chitosan-based nanoparticles as CpG ODN carriers
Preparation, characterization, and evaluation of chitosan-based nanoparticles as CpG ODN carriers
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DOI：
10.1080/13102818.2019.1578690
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发表时间：
2019-01-01
影响因子：
1.4
通讯作者：
Yamazaki, Tomohiko
中科院分区：
文献类型：
--
作者：
Iswanti, Febriana Catur;Nurulita, Indah;Yamazaki, Tomohiko
基于壳聚糖的颗粒由于其在组织中的粘膜粘附性、生物相容性、低毒性和在抗原转运中的有效性而成为理想的药物递送材料。如何有效地制备高稳定性的壳聚糖基微粒仍然是一个挑战。本研究以壳聚糖为载体制备了Toll样受体9配体--胞嘧啶-磷酸-鸟嘌呤寡脱氧核苷酸（cytosine-phosphate-guanine oligodeoxynucleotides，CpG ODNs）纳米粒，并将其用于诱导Th-2型向Th-1型免疫应答的转变。由于对DNA酶的低耐受性和游离CpG ODN进入细胞的较少摄取，所以开发带正电荷的载体对于将CpG ODN有效递送到免疫细胞中是必要的。采用离子凝胶法制备了壳聚糖纳米粒，并分别用动态光光谱仪和Zeta电位仪测定了纳米粒的粒径和表面电荷。合成的壳聚糖纳米粒在196 ° C处有两个分散峰，分散度为100%。29岁？nm和1.33 μ m。0.36？m，zeta电位为+3.3？0.4？MV.壳聚糖纳米颗粒对小鼠RAW 264巨噬细胞的毒性通过利用水溶性四唑盐WST-8的甲瓒染料测定来测量。我们的壳聚糖纳米颗粒对RAW 264细胞没有细胞毒性。最后，我们使用人外周血单个核细胞评价了负载在壳聚糖纳米颗粒上的CpG ODNs的免疫刺激活性。CpG ODNs能显著提高IL-6和IFN-γ的分泌水平。与游离CpG ODN相比，这些结果表明壳聚糖纳米粒可以作为CpG ODNs的载体。
Chitosan-based particles are desirable materials for drug delivery because of their muco-adhesiveness in tissues, biocompatibility, low toxicity and effectiveness in antigen transport. It is still a challenge to prepare chitosan-based particles with high stability and by an effective method. Here we developed chitosan-based nanoparticles for the delivery of toll-like receptor 9 ligands, cytosine-phosphate-guanine oligodeoxynucleotides (CpG ODNs), which can induce the change from Th-2 type to Th-1 type immune response. Because of the low tolerance to DNase and less uptake into the cells of the free CpG ODNs, the development of positively charged carriers is necessary for the effective delivery of CpG ODNs into immune cells. Chitosan nanoparticles were prepared by ionic gelation, and the nanoparticles size and surface charge were measured by dynamic light spectroscopy and zeta potential analyzer, respectively. The synthesized chitosan nanoparticles showed two dispersion peaks, at 196???29?nm and 1.33???0.36??m, with a zeta potential of +3.3???0.4?mV. The toxicity of the chitosan nanoparticles to murine RAW264 macrophages was measured by formazan dye assay utilizing the water-soluble tetrazolium salt WST-8. Our chitosan nanoparticles exhibited no cytotoxicity to RAW264 cells. Finally, we evaluated the immunostimulatory activity of CpG ODNs loaded on chitosan nanoparticles using human peripheral blood mononuclear cells. CpG ODNs significantly enhanced the secretion level of interleukin-6 and interferon-? by cells compared to the free CpG ODNs. These results indicated that chitosan nanoparticles could be a good candidate for the delivery of CpG ODNs.