Abstract

Stem cell tracking in cellular therapy and regenerative medicine is an urgent need, superparamagnetic iron oxide nanoparticles (IONPs) could be used as contrast agents in magnetic resonance imaging (MRI) that allows visualization of the implanted cells ensuring they reach the desired sites in vivo. Herein, we report the study of the interaction of 3,4-dihydroxyhydrocinnamic acid (DHCA) functionalized IONPs that have desirable properties for T2 - weighted MRI, with bone marrow-derived primary human mesenchymal stem cells (hMSCs). Using the multiparametric high-content imaging method, we evaluate cell viability, formation of reactive oxygen species, mitochondrial health, as well as cell morphology and determine that the hMSCs are minimally affected after labelling with IONPs. Their cellular uptake is visualized by transmission electron microscopy (TEM) and Prussian Blue staining, and quantified using an iron specific colourimetric method. In vitro and in vivo studies demonstrate that these IONPs are biocompatible and can produce significant contrast enhancement in T2-weighted MRI. Iron oxide nanoparticles are detected in vivo as hypointense regions in the liver up to two weeks post injection using 9.4 T MRI. These DHCA functionalized IONPs are promising contrast agents for stem cell tracking by T2-weighted MRI as they are biocompatible and show no evidence of cytotoxic effects on hMSCs.

Highlights

  • In recent years, research on the development of stem cell therapy has intensified

  • Most studies have reported the biocompatibility of iron oxide nanoparticles (IONPs) with in vitro and in vivo studies, and it is generally accepted within the scientific community that these are safer materials to use in comparison to other magnetic resonance imaging (MRI) contrast agents, such as gadolinium-based nanomaterials for example[11,12,13]

  • This study allowed us to determine the suitability of iron oxide nanoparticles synthesized as potential MRI contrast agents

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Summary

Introduction

Research on the development of stem cell therapy has intensified. The potential to use stem cells (SC) in tissue engineering and regenerative medicine is promising, as their use has already been implemented in a few human clinical trials[1,2,3,4]. Most studies have reported the biocompatibility of IONPs with in vitro and in vivo studies, and it is generally accepted within the scientific community that these are safer materials to use in comparison to other MRI contrast agents, such as gadolinium-based nanomaterials for example[11,12,13] Even though they are widely used and accepted, it is still necessary to assess the toxicity of newly developed IONPs and it is difficult to obtain a consensus amongst researchers on the methods used for determining their toxicity. As highlighted by Paul Weiss et al in ACS Nano editorial in November 201614, the lack of standardization when it comes to the study of nanoparticles remains an obstacle to their potential use in biomedical applications To this date, assessment of toxicity is routinely done through colourimetric assays such as MTT or MTS assays, it has been demonstrated that there is some interaction of dyes with IONPs and this leads to false positive results[15]. This allowed us to confirm the in vivo potential of the IONPs as safe and biocompatible T2-weighted MRI contrast agents

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