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Light and electron microscopic detection of inflammation-targeting liposomes encapsulating high-density colloidal gold in arthritic mice

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Abstract

Objective

We have previously demonstrated the efficient and time-dependent transvascular localization of Sialyl Lewis X (SLX)-liposomes to inflammatory sites, but the final target of the SLX-liposomes remained uncertain. The aim of this study was to identify the target cells of the liposomes within the inflamed joints of collagen antibody-induced arthritis (CAIA) model mice.

Methods

SLX-liposomes and unlabeled liposomes encapsulating high-density colloidal gold were administered intravenously into the caudal vein of CAIA mice on day 5 after induction of arthritis when the inflammatory score was maximal (n = 6 per group). Six hours or 24 h after liposome administration, animals were euthanized and hind limbs and ankles were excised without perfusion. After fixation, synovial tissues were examined by light microscopy after silver enhancement of colloidal gold or by transmission electron microscopy.

Results

Silver-enhanced signals were detected within the cells around E-selectin-positive blood vessels in the synovium of the SLX-liposome group. These cells were positive for the macrophage/monocyte marker F4/80 or neutrophil marker Ly-6G. Transmission electron microscopy detected the colloidal gold signals together with liposome-like structures within the phagosomes of synovial macrophages. Transmission electron microscopy and energy dispersive X-ray spectrometry could determine gold elements in the lysosomes of synovial macrophages.

Conclusions

The results of the current study demonstrate that SLX-liposomes primarily targeting E-selectin in activated endothelial cells could potentially deliver their contents into inflammatory cells around synovial blood vessels in arthritic joints.

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Acknowledgments

We thank Dr. Matsukawa for kindly providing anti-Ly-6G antibodies, Drs. Hirohata and Yonezawa for fruitful discussions, Dr. Ogawa for assistance with animal experiments, Mr. Urata for assistance with EDS analysis, and Drs. Otani, Hirai and Mr. Minematsu (Katayama Chemical Co. Ltd., Osaka, Japan) for assistance with liposome preparations. This work was supported in part by a Grant-in-Aid for Scientific Research on Innovative Areas (No. 24107516) to T.O., a Grant-in-Aid for Scientific Research (No. 23592215) to K.N., and a Grant-in-Aid for Challenging Exploratory Research (No. 24659590) to Y.N. from the Ministry of Education, Culture, Sports, Science, and Technology (MEXT) of Japan.

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Authors and Affiliations

  1. Department of Molecular Biology and Biochemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, 700-8558, Japan

    Ami Maehara, Emi Matsumoto, Yoshifumi Ninomiya & Toshitaka Oohashi

  2. Faculty of Physical Education, International Pacific University, Okayama, 709-0863, Japan

    Ami Maehara

  3. Department of Human Morphology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, 700-8558, Japan

    Keiichiro Nishida & Aiji Ohtsuka

  4. Central Research Laboratory, Okayama University Medical School, Okayama, 700-8558, Japan

    Masumi Furutani

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  1. Ami Maehara
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Correspondence to Toshitaka Oohashi.

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Maehara, A., Nishida, K., Furutani, M. et al. Light and electron microscopic detection of inflammation-targeting liposomes encapsulating high-density colloidal gold in arthritic mice. Inflamm. Res. 63, 139–147 (2014). https://doi.org/10.1007/s00011-013-0682-4

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  • DOI: https://doi.org/10.1007/s00011-013-0682-4

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