Study Of Superparamagnetic Iron Oxide Nanoparticles Modified With 2-DG Targeting Rhabdomyosarcoma Cell

Purpose?1?Prepare Fe₃O₄@OA NPs ? Fe₃O₄@OA-OA₂-PEG NPs and Fe₃O₄@OA-OA₂-PEG-DG NPs,investigate the characterizations.?2?Compare the rhabdomyosarcoma A-673 cells uptake of Fe₃O₄@OA-OA₂-PEG-DGNPs?Fe₃O₄@OA-OA₂-PEG NPs and Fe₃O₄@OA-OA₂-PEG-DG NPs in the presence of 4.5 g / L D-glucose,to detect cells targeting uptake of Fe₃O₄@OA-OA₂-PEG-DG NPs.Method?1?Fe₃O₄@OA NPs were synthesized by thermal decomposition,Fe₃O₄@OA-OA₂-PEG NPs and Fe₃O₄@OA-OA₂-PEG-DG NPs were prepared by surface modification.Observe the shape?size and dispersibility of the nanoparticles under the transmission electron microscope?TEM?,detect the magnetic properties of nanoparticles by vibrating sample magnetometer?VSM?,identify surface coating formation on nanoparticles via thermogravimetric analysis?TGA?.?2?0.3 ?mol/m LFe₃O₄@OA-OA₂-PEG-DG NPs?Fe₃O₄@OA-OA₂-PEG NPs and Fe₃O₄@OA-OA₂-PEG-DG NPs in the presence of 4.5 g/L D-glucose were incubated with rhabdomyosarcoma A-673 cells for 10 min?30 min?1 h and 2 h,detect cells targeting absorption with prussian blue staining,observe T2 WI sequence ? SWI sequence signal intensity and multiple echo sequence T2 values by MRI scanning,measure cell iron content by atomic absorption spectroscopy.Results?1?The morphology of Fe₃O₄@OA NPs were like cube,the average core particle size was about 10.4 ± 0.8 nm,Fe₃O₄@OA NPs were monodisperse in hexane,Fe₃O₄ @OA-OA₂-PEG NPs were stable and well dispersed in aqueous solution.The saturation magnetization of Fe₃O₄@OA NPs and Fe₃O₄@OA-OA₂-PEG NPs was 81.2± 5.3 emu/g? 59.7 ± 4.8 emu/g,respectively.TGA indicated OA and PEG were bound to the surface of Fe 3O 4 nanoparticles.?2?Prussian blue staining showed,incubating with Fe₃O₄@OA-OA₂-PEG-DG NPs and Fe₃O₄@OA-OA₂-PEG NPs for 10 min?30 min,there were little blue Granules inside the cells;at 1 h,blue Granules inside the cells of Fe₃O₄@OA-OA₂-PEG-DG NPs group increased;after 2 h,there were siginificant cells uptake of Fe₃O₄@OA-OA₂-PEG-DG NPs more than Fe₃O₄ @OA-OA₂-PEG NPs,which can be inhibited by glucose.?3?MRI T2 WI showed,after incubating with A-673 cell for 10 min?30 min,the signal intension of Fe₃O₄@OA-OA₂-PEG-DG NPs group?Fe₃O₄@OA-OA₂-PEG NPs group and competitive inhibition group did not decreased obviously;at 1 h,the signal intension of the three groups decreased,Fe₃O₄ @OA-OA₂-PEG-DG NPs group was slightly lower than the other two groups;at 2 h,the signal intension of Fe₃O₄@OA-OA₂-PEG-DG NPs group reduced highly,significantly lower than Fe₃O₄@OA-OA₂-PEG NPs group and competitive inhibition group.?4?MRI multiple echo sequence showed,after incubating with A-673 cell for 10 min ? 30 min,there was no statistic difference of T2 value between Fe₃O₄@OA-OA₂-PEG-DG NPs group and Fe₃O₄@OA-OA₂-PEG NPs group ? Fe₃O₄@OA-OA₂-PEG-DG NPs group and competitive inhibition group;at 1 h ? 2 h,the difference of T2 value between Fe₃O₄@OA-OA₂-PEG-DG NPs group and Fe₃O₄@OA-OA₂-PEG NPs group ? Fe₃O₄@OA-OA₂-PEG-DG NPs group and competitive inhibition group was prominent.?5?MRI SWI showed,the signal intension of Fe₃O₄@OA-OA₂-PEG-DG NPs group ? Fe₃O₄@OA-OA₂-PEG NPs group and competitive inhibition group all declined gradually with the extension of incubation time.At any time,the signal intension of Fe₃O₄@OA-OA₂-PEG-DG NPs group was lower than the two other groups.?6?Atomic absorption spectroscopy revealed,at different time,the celluar iron amount of Fe₃O₄@OA-OA₂-PEG-DG NPs group was higher than that of Fe₃O₄@OA-OA₂-PEG NPs group and competitive inhibition group.In addition,there was statistic difference between Fe₃O₄@OA-OA₂-PEG-DG NPs group and the two other groups at any time.Conclusion?1?Nanoparticles were successfully modified with oleic acid?OA?and polyethylene glycol?PEG?on the surface,had good stability and water solubility.?2?Fe₃O₄@OA-OA₂-PEG-DG NPs targeted the highly glut-expressing A-673 cells well,PEG can resist cells nonspecific adhesion,which realize markers' specific identification for the receptors better,and make Fe₃O₄@OA-OA₂-PEG-DG NPs target tumor cell more precisely,Fe₃O₄@OA-OA₂-PEG-DG NPs is expect to be a new type of molecular magnetic resonance probe for malignant tumor targeting.SWI sequence is sensitive to low concentration of superparamagnetic iron oxide nanoparticles,which is expected to provide effective detection method for low concentration of contrast agents targeting tumors in vivo.