Huang K-J, Lee C-Y, Lin Y-C, Lin C-Y, Perevedentseva E, Hung S-F, Cheng C-L
Department of Life Sciences, National Dong Hwa University, 1, Sec. 2 Da Hsueh Rd, Shoufeng, Hualien, 97401, Taiwan.
Institute of Biologicals, Development Center for Biotechnology (DCB), New Taipei City, 22180, Taiwan.
J Biophotonics. 2017 Oct;10(10):1315-1326. doi: 10.1002/jbio.201600202. Epub 2017 Jan 9.
The applications of nanodiamond as drug delivery and bio-imaging can require the relinquishing ND-drug conjugate via blood flow, where interaction with immune cells may occur. In this work, we investigated the ND penetration in macrophage and the immune response using the tissue-resident murine macrophages (RAW 264.7). Confocal fluorescence imaging, immunofluorescence analysis of nuclear translocation of interferon regulatory factor IRF-3 and transcriptional factor NF-κΒ, analysis of pro-inflammatory cytokines production IL-1β, IL-6 IL-10 with a reverse transcription-polymerase chain reaction technique were applied. The TNF-α factor production has been studied both in vitro at ND interaction with the macrophage and in vivo after ND injection in the mice blood system using immunoassay. The macrophage antibacterial function was estimated through E. coli bacterial colony formation. ND didn't stimulate the immune response and functionality of the macrophage was not altered. Using MTT test, ND was found negligibly cytotoxic to macrophages. Thus, ND can serve as a biocompatible platform for bio-medical applications. Left: Graphic representation of Nanodiamond internalization in macrophage. Right: (a) Fluorescence images of lysosomes, (b) nanodiamond and (c) merged image of nanodiamond internalization in macrophage.
纳米金刚石在药物递送和生物成像方面的应用可能需要通过血流释放纳米金刚石-药物偶联物，在此过程中可能会与免疫细胞发生相互作用。在这项工作中，我们使用组织驻留小鼠巨噬细胞（RAW 264.7）研究了纳米金刚石在巨噬细胞中的渗透情况以及免疫反应。应用了共聚焦荧光成像、干扰素调节因子IRF-3和转录因子NF-κB核转位的免疫荧光分析、采用逆转录-聚合酶链反应技术分析促炎细胞因子IL-1β、IL-6、IL-10的产生。使用免疫测定法研究了在体外纳米金刚石与巨噬细胞相互作用时以及在体内纳米金刚石注射到小鼠血液系统后TNF-α因子的产生情况。通过大肠杆菌菌落形成评估巨噬细胞的抗菌功能。纳米金刚石未刺激免疫反应，巨噬细胞的功能也未改变。使用MTT试验发现纳米金刚石对巨噬细胞的细胞毒性可忽略不计。因此，纳米金刚石可作为生物医学应用的生物相容性平台。左图：巨噬细胞中纳米金刚石内化的示意图。右图：（a）溶酶体的荧光图像，（b）纳米金刚石，（c）巨噬细胞中纳米金刚石内化的合并图像。
