Gadolinium labelled nanoliposomes as the platform for MRI theranostics: in vitro safety study in liver cells and macrophages

Pavlína Šimečková,Jiřina Procházková,František Hubatka,Ondrej Kováč,Josef Slavík,Kateřina Pěnčíková,Josef Mašek,Stuart Macaulay,Miroslav Machala,Jana Stráská,Pavel Kulich,Milan Raška,Robert Mikulík,Dominik Hrebík,Jiří Neča,Jaroslav Turánek,Pavlína Turánek Knötigová,Jan Kotouček,Pavel Diviš

doi:10.1038/s41598-020-60284-z

Abstract

Gadolinium (Gd)–based contrast agents are extensively used for magnetic resonance imaging (MRI). Liposomes are potential nanocarrier–based biocompatible platforms for development of new generations of MRI diagnostics. Liposomes with Gd–complexes (Gd–lip) co–encapsulated with thrombolytic agents can serve both for imaging and treatment of various pathological states including stroke. In this study, we evaluated nanosafety of Gd–lip containing PE-DTPA chelating Gd+3 prepared by lipid film hydration method. We detected no cytotoxicity of Gd–lip in human liver cells including cancer HepG2, progenitor (non–differentiated) HepaRG, and differentiated HepaRG cells. Furthermore, no potential side effects of Gd–lip were found using a complex system including general biomarkers of toxicity, such as induction of early response genes, oxidative, heat shock and endoplasmic reticulum stress, DNA damage responses, induction of xenobiotic metabolizing enzymes, and changes in sphingolipid metabolism in differentiated HepaRG. Moreover, Gd–lip did not show pro–inflammatory effects, as assessed in an assay based on activation of inflammasome NLRP3 in a model of human macrophages, and release of eicosanoids from HepaRG cells. In conclusion, this in vitro study indicates potential in vivo safety of Gd–lip with respect to hepatotoxicity and immunopathology caused by inflammation.

Highlights

Gadolinium (Gd)–based contrast agents are extensively used for magnetic resonance imaging (MRI)
Gadolinium containing liposome (Gd–lip) preparations were represented by unilamellar liposomes with low polydispersity as demonstrated by dynamic light scattering (DLS) and cryo–transmission electron microscopy (TEM) methods
We found no changes in any tested parameters, which indicates that neither ctrl–lip nor Gd–lip induced activation of inflammasome in THP1 cells and production of pro–inflammatory cytokines in differentiated HepaRG

Summary

Introduction

Gadolinium (Gd)–based contrast agents are extensively used for magnetic resonance imaging (MRI). Liposomes can serve as diagnostic as well as theranostic agents for imaging and treatment of various pathological states and illnesses such as cancer, ischemic stroke and vasculature of different organs including liver and spleen[6,7]. Different surface coating of liposomes including polyethylene glycol, hyaluronic acid and polysaccharides[9,10,11] or N-(2-hydroxypropyl) methacrylamide polymers[12] are suitable to achieve targeting and long–circulation properties of liposomes intended as carriers for contrast agents. By incorporating both PE-DTPA (Gd) and a near infrared dye, liposomal formulations can be used for multimodal imaging[13]. A more detailed study of potential toxicological effects of lipid–based Gd+3 formulations on different organs and cell types including liver cells and immune cells is of importance for a future clinical application in patients

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Results

Conclusion