Department of Biomaterials Engineering, Kangwon National University , Chuncheon 200-701, Republic of Korea.
ACS Nano. 2013 Nov 26;7(11):9771-9. doi: 10.1021/nn403275p. Epub 2013 Oct 9.
Electrodeposited nanorods serving as multivalent bridges were fabricated and surface-decorated with ligands for immune cells. Gold and nickel solutions were sequentially electrodeposited on nanoporous anodized disc templates and the template was dissolved to retrieve bisegmented nanorods with different lengths. Gold and nickel segmented nanorods were surface-immobilized with mannose and RGD peptides to prepare immune-cell recruiting nanorods. Surface-functionalization of nanorods were confirmed by fluorescence-labeling of each ligands and confocal microscopy. Dendritic cells and T cells were co-incubated with the surface-functionalized nanorods, and the proximity between the nanorods and the immune cells was visualized by variable pressure scanning electron microscopy and confocal microscopy. The long nanorods were associated with the immune cells, whereas the shorter nanorods were rather endocytosed by cells, suggesting a feasibility of the longer nanorods as bridging for the cells. Cytokine releases from the immune cells were monitored by cultivating lipopolysaccharide-activated dendritic cells with T cells. Interleukine-2 and interferon-γ release profiles showed a strong correlation with the length of the nanorod, where the 4 μm nanorods induced the highest levels of cytokine release compared to 1 or 2 μm nanorods. Thus, we concluded that the proximity of the immune cells increased by bridging the immune cells with the nanobridging system, which subsequently increased cytokine release by facilitating the antigen presentation process.
作为多价桥的电沉积纳米棒被制造出来，并在表面用免疫细胞的配体进行修饰。金和镍溶液依次在纳米多孔阳极氧化铝模板上电沉积，然后溶解模板以回收具有不同长度的双节纳米棒。金和镍分段纳米棒被甘露糖和 RGD 肽表面固定化，以制备免疫细胞招募纳米棒。通过对每个配体进行荧光标记和共焦显微镜观察，证实了纳米棒的表面功能化。将表面功能化的纳米棒与树突状细胞和 T 细胞共孵育，通过变压扫描电子显微镜和共焦显微镜观察纳米棒与免疫细胞的接近程度。长纳米棒与免疫细胞相关联，而较短的纳米棒则被细胞内吞，这表明较长的纳米棒作为细胞桥的可行性。通过用 T 细胞培养脂多糖激活的树突状细胞来监测免疫细胞释放的细胞因子。白细胞介素-2 和干扰素-γ的释放谱与纳米棒的长度呈强相关，其中 4μm 的纳米棒与 1μm 或 2μm 的纳米棒相比，诱导细胞因子释放的水平最高。因此，我们得出结论，通过纳米桥接系统将免疫细胞桥接起来，可以增加免疫细胞的接近程度，从而通过促进抗原呈递过程增加细胞因子的释放。
