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Biological characterization of a novel hybrid copolymer carrier system based on glycogen

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Abstract

The effective drug delivery systems for cancer treatment are currently on high demand. In this paper, biological behavior of the novel hybrid copolymers based on polysaccharide glycogen were characterized. The copolymers were modified by fluorescent dyes for flow cytometry, confocal microscopy, and in vivo fluorescence imaging. Moreover, the effect of oxazoline grafts on degradation rate was examined. Intracellular localization, cytotoxicity, and internalization route of the modified copolymers were examined on HepG2 cell line. Biodistribution of copolymers was addressed by in vivo fluorescence imaging in C57BL/6 mice. Our results indicate biocompatibility, biodegradability, and non-toxicity of the glycogen-based hybrid copolymers. Copolymers were endocyted into the cytoplasm, most probably via caveolae-mediated endocytosis. Higher content of oxazoline in polymers slowed down cellular uptake. No strong colocalization of the glycogen-based probe with lysosomes was observed; thus, it seems that the modified externally administered glycogen is degraded in the same way as an endogenous glycogen. In vivo experiment showed relatively fast biodistribution and biodegradation. In conclusion, this novel nanoprobe offers unique chemical and biological attributes for its use as a novel drug delivery system that might serve as an efficient carrier for cancer therapeutics with multimodal imaging properties.

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Abbreviations

7-AAD:

7-aminoactinomycin D

DMSO:

dimethyl sulfoxide

DOTA:

1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid

EPR:

enhanced permeability and retention

ESI-MS:

electrospray ionization mass spectrometry

FITC:

fluorescein isothiocyanate

GG:

glycogen

HBSS:

Hanks’ Balanced Salt Solution

LGS:

liver glycogen synthase

MBCD:

methyl-β-cyclodextrin

MEM:

Minimum Essential Medium

MRI:

magnetic resonance imaging

NHS ester:

N-hydroxysuccinimide ester

ROI:

region of interest

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Acknowledgements

This work was supported by the Grant Agency of the Ministry of Health, Czech Republic [grant number 15-25781a], the Czech Science Foundation [grant number 13-08336S], the Charles University in Prague (Faculty of Science), and project GA UK [grant number 282216], and the confocal microscopy part (M. Pařízek) was supported by the Czech Science Foundation [grant number P108/12/1168]. We would like to thank Alena Sekerková for help with flow cytometry measurements.

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Authors and Affiliations

  1. Institute for Clinical and Experimental Medicine (IKEM), Videnska 1958/9, 140 21, Prague 4, Czech Republic

    Markéta Jirátová, Jan Kovář, Andrea Gálisová & Daniel Jirák

  2. Department of Physiology, Faculty of Science, Charles University in Prague, Viničná 7, 128 44, Praha 2, Czech Republic

    Markéta Jirátová

  3. Institute of Macromolecular Chemistry of the Academy of Sciences of the Czech Republic, Heyrovsky Sq. 2, 162 06, Prague 6, Czech Republic

    Aneta Pospíšilová, Maria Rabyk & Martin Hrubý

  4. Institute of Physiology of the Academy of Sciences of the Czech Republic, Videnska 1083, 142 20, Prague 4, Czech Republic

    Martin Pařízek

  5. Institute of Biophysics and Informatics, 1st Medicine Faculty Charles University, Salmovská 1, 12000, Prague 2, Czech Republic

    Andrea Gálisová & Daniel Jirák

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  1. Markéta Jirátová
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  2. Aneta Pospíšilová
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Correspondence to Daniel Jirák.

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All protocols were approved by the Ethical Committee of the Institute for Clinical and Experimental Medicine and the experiments were carried out in accordance with the European Communities Council Directive (86/609/EEC). All institutional and national guidelines for the care and use of laboratory animals were followed.

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The authors declare that they have no conflict of interest.

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Jirátová, M., Pospíšilová, A., Rabyk, M. et al. Biological characterization of a novel hybrid copolymer carrier system based on glycogen. Drug Deliv. and Transl. Res. 8, 73–82 (2018). https://doi.org/10.1007/s13346-017-0436-x

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