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In Vitro and In Vivo Evaluation of Fluorescently Labeled Borocaptate-Containing Liposomes

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Abstract

Boron neutron capture therapy (BNCT), a binary cancer therapeutic modality, has moved to a new phase since development of accelerator-based neutron sources and establishment of BNCT centers in Finland and Japan. That stimulated efforts for better boron delivery agent development. As liposomes have shown effective boron delivery properties and sufficient tumor retention, fluorescent liposome labelling may serve as a rapid method to study initial ability of newly synthesized liposomes to be captured by tumor cells prior to experiments on boron accumulation and neutron irradiation. In this work, we studied the accumulation and biodistribution of pegylated liposomes with encapsulated borocaptate (BSH) and a fluorescent label (Nile Red) in U87 (human glioblastoma), SW-620 (human colon carcinoma), SK-MEL-28 (human melanoma), FetMSC (mesenchymal human embryo stem cells), and EMBR (primary embryocytes) cell lines as well as an orthotopic xenograft model of U87 glioma in SCID mice. Results indicate that fluorescent microscopy is effective at determining the intracellular localization of the liposomes using a fluorescent label. The synthesized, pegylated liposomes showed higher accumulation in tumors compared to normal cells, with characteristic concentration peaks in SW-620 and U87 cell lines, and provided in vivo tumor selectivity with several-fold higher tumor tissue fluorescence at the 6-h timepoint.

Fluorescent images of U-87 glioma cells after 24 hours of incubation with BSH-containing liposomes labeled with lipophilic Nile Red (red color)and water-soluble FITC-Dextran (green color); cell nuclei in blue color (DAPI-staining) (×400). Scale bar is 50 μm. Fluorescent labelling serves as anexpress method to study liposome delivery efficiency prior to boron accumulation evaluation and BNCT irradiation experiments.

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Authors and Affiliations

  1. Laboratory of Medical and Biological Problems of BNCT, Novosibirsk State University, Novosibirsk, Russian Federation

    Vladimir Kanygin, Alexander Zaboronok, Evgenii Zavjalov, Rinat Mukhamadiyarov, Aleksandr Kichigin, Anna Kasatova, Ivan Razumov & Roman Sibirtsev

  2. Faculty of Medicine, University of Tsukuba, Tsukuba, Ibaraki, Japan

    Alexander Zaboronok & Bryan J. Mathis

  3. Department of Neurosurgery, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8575, Japan

    Alexander Zaboronok

  4. Laboratory of Ultrastructural Research, Research Institute of Clinical and Experimental Lymphology – Branch of the Institute of Cytology and Genetics SB RAS, Novosibirsk, Russian Federation

    Iuliia Taskaeva

  5. Laboratory of BNCT, Novosibirsk State University, Novosibirsk, Russian Federation

    Iuliia Taskaeva

  6. Budker Institute of Nuclear Physics, Novosibirsk, Russian Federation

    Iuliia Taskaeva & Anna Kasatova

  7. Center for Genetic Resources of Laboratory Animals, Institute of Cytology and Genetics SB RAS, Novosibirsk, Russian Federation

    Evgenii Zavjalov & Ivan Razumov

  8. Research Institute for Complex Issues of Cardiovascular Diseases SB RAS, Kemerovo, Russian Federation

    Rinat Mukhamadiyarov

Authors
  1. Vladimir Kanygin
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  2. Alexander Zaboronok
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  3. Iuliia Taskaeva
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  4. Evgenii Zavjalov
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  6. Aleksandr Kichigin
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  10. Bryan J. Mathis
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Contributions

Conceptualization: Iuliia Taskaeva, Rinat Mukhamadiyarov, Evgenii Zavjalov and Ivan Razumov; Data acquisition, analysis and interpretation: Evgenii Zavjalov, Ivan Razumov, Roman Sibirtsev, Rinat Mukhamadiyarov, Iuliia Taskaeva, Aleksandr Kichigin, Anna Kasatova, Alexander Zaboronok; Funding and resources acquisition: Vladimir Kanygin, Rinat Mukhamadiyarov, Evgenii Zavjalov and Ivan Razumov; Project administration and supervision: Vladimir Kanygin and Evgenii Zavjalov; Writing original draft, review and editing: Alexander Zaboronok, Aleksandr Kichigin and Bryan J. Mathis.

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Correspondence to Alexander Zaboronok.

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For this type of study, the ethical approval was not required, as all manipulations with animals were carried out in the specialized animal center in compliance with research practices in accordance with the directives of the European Community (86/609 / EEC).

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Kanygin, V., Zaboronok, A., Taskaeva, I. et al. In Vitro and In Vivo Evaluation of Fluorescently Labeled Borocaptate-Containing Liposomes. J Fluoresc 31, 73–83 (2021). https://doi.org/10.1007/s10895-020-02637-5

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