Superparamagnetic iron oxide nanoparticles may affect endothelial progenitor cell migration ability and adhesion capacity.
OBJECTIVE
Cell labeling with superparamagnetic iron oxide (SPIO) nanoparticles enables non-invasive tracking of transplanted cells. The aim of this study was to investigate whether SPIO nanoparticles have an effect on endothelial progenitor cell (EPC) functional activity and the feasibility of a protocol for labeling swine- and rat-origin EPC using SPIO nanoparticles at an optimized low dosage.


METHODS
EPC were isolated from the peripheral blood of swine and bone marrow of rat and characterized. After ex vivo cultivation, EPC were labeled with SPIO nanoparticles (to make a series of final concentrations, 50, 100, 200 and 400 microg/mL) or vehicle control. We also investigated the long-term effects of 200 microg/mL SPIO nanoparticles on EPC (4, 8, 12 and 16 days after labeling). The labeling efficiency was tested through Prussian blue (PB) staining and the intracellular iron uptake was also measured quantitatively and confirmed. EPC proliferation and migration were determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and transwell chamber assay, respectively. An EPC adhesion assay was performed by replating the cells on fibronectin-coated dishes and then counting the adherent cells. EPC apoptosis was evaluated using an Annexin V-FITC apoptosis kit.


RESULTS
SPIO nanoparticles impaired EPC migration and promoted EPC adhesion. EPC proliferation and apoptosis were not affected. SPIO nanoparticles could label EPC efficiently at 200 microg/mL overnight without significantly affecting EPC functional activity.


CONCLUSIONS
SPIO nanoparticles impaired the EPC migration ability and promoted the EPC adhesion capacity. EPC could be labeled efficiently at an appropriate concentration (200 microg/mL) without significantly affecting their functional activity.
Yang
Jin-Xiu
JX
Department of Cardiology, The First Affiliated Hospital, Medical School of Zhejiang University, Hangzhou, China.
Tang
Wei-Liang
WL
Wang
Xing-Xiang
XX
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D003911:Dextrans; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D005295:Ferrocyanides / Q000378:metabolism; D052203:Ferrosoferric Oxide / Q000494:pharmacology*; D007501:Iron / Q000378:metabolism; D058185:Magnetite Nanoparticles; D053758:Nanoparticles / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013194:Staining and Labeling; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000648:ultrastructure; D034421:Sus scrofa
D003911:Dextrans; D005295:Ferrocyanides; D058185:Magnetite Nanoparticles; D007501:Iron; C065507:ferumoxides; C000170:ferric ferrocyanide; D052203:Ferrosoferric Oxide