In vivo bioimaging of macrophages using fluorescent microcapsules as a model carrier for the delivery of high-molecular cargo
Anastasia Yu. Sapach 1, Olga A. Sindeeva 1, Mikhail V. Nesterchuk 1,2 Alexandra A. Tsitrina 2, Ekaterina S. Prikhozhdenko 3, Roman A. Verkhovskii 3, Yuri V. Kotelevtsev 1,2, Gleb B. Sukhorukov 1,4
1 Skolkovo Institute of Science and Technology, Skolkovo Innovation Center, 143025, Moscow, Russia
2 Koltzov Institute of Developmental Biology of Russian Academy of Sciences, Moscow, 119334, Russia
3 Saratov State University, Saratov 410012, Russia
4 Queen Mary University of London, London E1 4NS, U.K.
Abstract
The "Trojan horse" is a hot topic of using the organism's own cells for delivery of drug carriers in cell therapy. So, macrophages mediated delivery of synthetic microcapsules based drug carriers is a very promising strategy to enhance control over the distribution of cells and cargo and improve targeting.
The aim of the study was to develop an approach for in vivo bioimaging of macrophages using microcapsules based on biocompatible polyelectrolytes and containing two fluorescent labels as a cargo (BSA-Cy7 in the capsule interior) and a layer (RITC-BSA in the shell).
The capsules were fabricated by layer-by-layer (LbL) method and characterized with SEM and CLSM techniques.
In this work, we investigate the effects of microcapsules on the viability, motility and proliferation of immortalised (RAW 264.7) and primary cells (BMDM) macrophages in vitro using the MTT test, CLSM, flow cytometry. In vivo and ex vivo, we compare the biodistribution and localisation of both macrophage types with microcapsules using the IVIS SpectrumCT In Vivo Imaging System. It was found that the ratio of 5 capsules per cell does not affect both types of macrophages functionality in vitro. In addition, the capsules allow us to safely detect both types of macrophages injected into the tail vein during 24 h in vivo and to evaluate the localization in organs and tissues in more detail ex vivo.
Thus, fluorescent multilayer polymer microcapsules can be used as a promising approach for tracking and loading macrophage mediated delivery systems.
This work was supported by Russian Science Foundation (project no. 19-75-10043).
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Anastasiia Sapach
Skolkovo Institute of Science and Technology, Skolkovo Innovation Center, Moscow, Russia
Russia
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