Phagocytosis and immune response studies of Macrophage‐Nanodiamond Interactions in vitro and in vivo

• Published in: Journal of biophotonics Vol. 10; no. 10; pp. 1315 - 1326
• Main Authors: Huang, K.‐J., Lee, C.‐Y., Lin, Y.‐C., Lin, C.‐Y., Perevedentseva, E., Hung, S.‐F., Cheng, C.‐L.
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY‐VCH Verlag 01-10-2017
• Subjects: Animals; Biological Transport; bio‐labeling; cytotoxicity; immunological response; macrophage; Macrophages - cytology; Macrophages - drug effects; Macrophages - immunology; Macrophages - metabolism; Mice; nanodiamond; Nanodiamonds - toxicity; Phagocytosis - drug effects; RAW 264.7 Cells; nanodiamond; macrophage; immunological response; bio-labeling; cytotoxicity
• ISSN: 1864-063X; 1864-0648
• DOI: 10.1002/jbio.201600202

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.
The applications of nanodiamond (ND) 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). It was shown that 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.