title: Cellular level loading and heating of superparamagnetic iron oxide nanoparticles.
abstract: Superparamagnetic iron oxide nanoparticles (NPs) hold promise for a variety of biomedical applications due to their properties of visualization using magnetic resonance imaging (MRI), heating with radio frequency (rf), and movement in an external magnetic field. In this study, the cellular loading (uptake) mechanism of dextran- and surfactant-coated iron oxide NPs by malignant prostate tumor cells (LNCaP-Pro5) has been studied, and the feasibility of traditional rf treatment and a new laser heating method was evaluated. The kinetics of cell loading was quantified using magnetophoresis and a colorimetric assay. The results showed that loading of surfactant-coated iron oxide NPs with LNCaP-Pro5 was saturable with time (at 24 h) and extracellular concentration (11 pg Fe/cell at 0.5 mg Fe/mL), indicating that the particles are taken up by an "adsorptive endocytosis" pathway. Dextran-coated NPs, however, were taken up less efficiently (1 pg Fe/cell at 0.5 mg Fe/mL). Loading did not saturate with concentration suggesting uptake by fluid-phase endocytosis. Magnetophoresis suggests that NP-loaded cells can be held using external magnetic fields in microcirculatory flow velocities in vivo or in an appropriately designed extracorporeal circuit. Loaded cells were heated using traditional rf (260A, 357 kHz) and a new laser method (532 nm, 7 ns pulse duration, 0.03 J/pulse, 20 pulse/s). Iron oxide in water was found to absorb sufficiently strongly at 532 nm such that heating of individual NPs and thus loaded cells (1 pg Fe/cell) was effective (<10% cell survival) after 30 s of laser exposure. Radio frequency treatment required higher loading (>10 pg Fe/cell) and longer duration (30 min) when compared to laser to accomplish cell destruction (50% viability at 10 pg Fe/cell). Scaling calculations show that the pulsed laser method can lead to single-cell (loaded with NPs) treatments (200 degrees C temperature change at the surface of an individual NP) unlike traditional rf heating methods which can be used only for bulk tissue level treatments. In a mixture of normal and NP-loaded malignant tumor cells, the malignant cells were selectively destroyed after laser exposure leaving the unloaded normal cells intact. These studies hold promise for applications in cell purification and sorting and extracorporeal blood treatments in vitro.
authors.affiliation: Department of Mechanical Engineering, University of Minnesota, 111 Church Street SE, Minneapolis, MN 55455, USA.
authors.forename: Venkat S
authors.initials: VS
authors.lastname: Kalambur
authors.forename: Ellen K
authors.initials: EK
authors.lastname: Longmire
authors.forename: John C
authors.initials: JC
authors.lastname: Bischof
mesh_terms: D000465:Algorithms; D045744:Cell Line, Tumor; D003124:Colorimetry; D005290:Ferric Compounds / Q000737:chemistry*; D006361:Heating / Q000379:methods*; D006801:Humans; D007700:Kinetics; D008279:Magnetic Resonance Imaging; D008280:Magnetics; D008297:Male; D053758:Nanoparticles / Q000737:chemistry*; D010316:Particle Size; D011471:Prostatic Neoplasms / Q000473:pathology / Q000628:therapy*; D013696:Temperature
chemical_list: D005290:Ferric Compounds; C000499:ferric oxide