The Effect of Uncoated SPIONs on hiPSC-Differentiated Endothelial Cells

Abstract

Background: Endothelial progenitor cells (EPCs) were indicated in vascular repair, angiogenesis of ischemic organs, and inhibition of formation of initial hyperplasia. Differentiation of endothelial cells (ECs) from human induced pluripotent stem cells (hiPSC)-derived endothelial cells (hiPSC-ECs) provides an unlimited supply for clinical application. Furthermore, magnetic cell labelling offers an effective way of targeting and visualization of hiPSC-ECs and is the next step towards in vivo studies. Methods: ECs were differentiated from hiPSCs and labelled with uncoated superparamagnetic iron-oxide nanoparticles (uSPIONs). uSPION uptake was compared between hiPSC-ECs and mature ECs isolated from patients by software analysis of microscopy pictures after Prussian blue cell staining. The acute and long-term cytotoxic effects of uSPIONs were evaluated by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay) and Annexin assay. Results: We showed, for the first time, uptake of uncoated SPIONs (uSPIONs) by hiPSC-ECs. In comparison with mature ECs of identical genetic background hiPSC-ECs showed lower uSPION uptake. However, all the studied endothelial cells were effectively labelled and showed magnetic properties even with low labelling concentration of uSPIONs. uSPIONs prepared by microwave plasma synthesis did not show any cytotoxicity nor impair endothelial properties. Conclusion: We show that hiPSC-ECs labelling with low concentration of uSPIONs is feasible and does not show any toxic effects in vitro, which is an important step towards animal studies.

3D bioprinting of nanoparticle-laden hydrogel scaffolds with enhanced antibacterial and imaging properties

“…Interestingly, there were no significant decreases in cell viability at day 7 for the NIH3T3 cells in contrast to the HUVEC results, suggesting greater tolerance of the fibroblast line against the NP-associated cytotoxic effects (as reported before ( Kong et al., 2011 ; Sahu et al., 2016 )). Of note, in both EC and fibroblast cells, clathrin and caveolae-mediated endocytosis are the common receptors used for the uptake of NPs, including the SPIONs ( Behzadi et al., 2017 ; Pottler et al., 2017 ; Rejman et al., 2004 ; Salingova et al., 2019 ). Thus, the SPION uptake in these cells can be regulated by manipulating these cell receptor pathways, as well as tuning the NP properties (e.g., size, protein corona, and surface charge) ( Behzadi et al., 2017 ; Rejman et al., 2004 ).…”

Nanotechnology-Assisted Cell Tracking

“…Prussian blue [48,49,51,[57][58][59]64,65,88,109,114,116,117,124,127,138,154,155,160,177,187,189,[209][210][211][212][213][214][215][216][217][218][219][220][221][222][223][224] Oil Red O [58,225] Alizarin red [225] Dyes for NP coating Propidium iodide (PI) [118] Nile red [89] Moreover, ex vivo checking, and quantification of cell uptake was performed by coating NPs with PI or Nile red dye. As example, NP coating with PI was successfully adopted to detect cells incorporating gold NPs through flow cytometry [118], whereas Nile red labeling of hybrid iron oxide/polymeric NPs was used to check in vitro uptake into macrophages [89].…”