Assessing cell-nanoparticle interactions by high content imaging of biocompatible iron oxide nanoparticles as potential contrast agents for magnetic resonance imaging

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.

In vitro toxicity screening of magnetite nanoparticles by applying mesenchymal stem cells derived from human umbilical cord lining

“…Moreover, we observed changes in cell morphology and loss of the continuous monolayer at the highest Fe 3 O 4 NP concentrations (≥50 μg/mL) in accordance with recent findings showing morphological alterations, characterized by the nuclear fragmentation (Periasamy et al, ), modification in cytoskeleton components (Dulinska‐Molak et al, ) and changes in the actin cytoskeleton (Hachani et al, ) depending on concentration and time of exposure.…”

“…To date, most in vitro Fe 3 O 4 NP toxicity information was derived from studies carried out on immortalized or tumorigenic cells (Coccini, Caloni, Ramírez Cando, & De Simone, ; Guadagnini, Moreau, Hussain, Marano, & Boland, ; Mahmoudi, Hofmann, Rutishauser, & Fink, ; Malvindi et al, ; Sanganeria, Sachar, et al, ). Recent studies have started to apply MSCs for Fe 3 O 4 NP toxicity evaluation (in terms of cytotoxicity, MSC differentiation capacity and iron uptake), which mostly derive from BM (Hachani et al, ; Harrison et al, ; Rosenberg et al, ), few from UC tissue (Dulinska‐Molak et al, ; Hu et al, ; Periasamy, Athinarayanan, Alhazmi, Alatiah, & Alshatwi, ) and only one from CL‐MSC (Sanganeria, Chandra, Bahadur, & Khanna, ).…”

Iron Oxide Nanoparticle-Induced Autophagic Flux Is Regulated by Interplay between p53-mTOR Axis and Bcl-2 Signaling in Hepatic Cells

“…Cells onto 96-well clean bottom plates at the density of 5000 cells per well and treated with different concentration of fluorescently-labelled NPs for 2, 6, 8, and 24 h. Afterwards, cells were washed with PBS three times and incubated with 0.1% TrypanBlue in order to quench the fluorescent signal of NPs attached to the cell surface. TrypanBlue is widely used as a quencher of FITC fluorescence and is excluded from viable cells [40][41][42]. The NPs uptake was assessed by measuring of fluorescent intensity (Ex: 485 nm; Em: 540 nm) using TECAN microplate reader SpectraFluor Plus (TECAN, Mannedorf, Switzerland).…”

A Modular Millifluidic Platform for the Synthesis of Iron Oxide Nanoparticles with Control over Dissolved Gas and Flow Configuration

“…Magnetic nanoparticles represent a class of nanomaterials of particular interest in the biomedical field [1][2][3], with potential applications in thermal tissue ablation [4,5], immunoassays [6,7], magnetic resonance imaging [8][9][10][11], magnetic particle imaging [12], drug delivery [13][14][15][16], pathogen diagnostic assays [17] and tissue repair [18]. For these biomedical applications, iron oxide nanoparticles (IONPs) still represent the most promising magnetic nanomaterials due to their biocompatibility [1,4].…”