The Duality of Iron Oxide Nanoparticles in Cancer Therapy: Amplification of Heating
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1 Supporting Information The Duality of Iron Oxide Nanoparticles in Cancer Therapy: Amplification of Heating Efficiency by Magnetic Hyperthermia and Photothermal Bimodal Treatment Ana Espinosa, Riccardo Di Corato, Jelena Kolosnjaj-Tabi, Patrice Flaud, Teresa Pellegrino, and Claire Wilhelm, * Laboratoire Matière et Systèmes Complexes, UMR 757, CNRS and University Paris Diderot, 7525 Paris cedex 13, France. Istituto Italiano di Tecnologia, I Genoa, Italy. 1
2 FIGURES T CAMERA ( C) Calibration mm MHT 52 khz T FLUO ( C) Figure S1. Calibration of the infrared (IR) camera temperature measurement. A suspension of iron oxide nanocubes (at [Fe] = 6 mm) was heated under a MHT protocol (52 khz and 25 mt) and temperature was recorded simultaneously using an optical fluorometer and the IR camera. The fluorometer probe was immersed in the iron oxide suspension and the IR camera focused on the surface of the sample. 4 3 LASER.3 W/cm 2 LASER.8 W/cm 2 T ( C) [Fe] mm Figure S2. Heat generation (temperature increase) by iron oxide nanocubes in suspension at high concentrations ([Fe] = -25 mm) under LASER mode after 5 min at.3 and.8 W/cm 2. 2
3 Figure S3. Laser attenuation as a function of iron concentration. Temperature elevation mapping of lateral view of samples under LASER mode at.3 W/cm 2. Measurements were performed outside the magnetic coil setup for [Fe] = 6, 25 and 1 mm after 1 s. Two areas were selected: area 1 (temperature integration over the entire sample area) and area 2 (temperature integration within the first layers 3 mm). A Absorbance [Fe] gallol-peg 25 mm 12 mm.8 mg/ml.4 mg/ml λ (nm) B T ( C) LASER.3 W/cm 2 LASER.8 W/cm [gallol-peg] mg/ml Figure S4. Contribution of the nanocubes surface coating (gallol-peg) to the photo-thermal properties. (A). UV-Vis-NIR absorbance spectra of the (gallol-peg) surface coating (in mg/ml) corresponding to iron oxide nanocubes samples at [Fe] = 12 and 25 mm. (B) Corresponding temperature increase after 5 min under LASER mode at.3 and.8 W/cm 2. 3
4 12 9 LASER.8 W/cm 2 T ( C) nm 88 nm 164 nm Figure S5. Temperature increase of iron oxide nanocubes suspension ([Fe] = 12 mm) after 3 s under LASER mode at.8 W/cm 2 and for three different laser wavelengths (68, 88 and 164 nm). SLP (W/g) khz H (mt) Figure S6. Heating efficiency of iron oxide nanocubes in suspension as a function of magnetic field. Heating capacity (SLP (W/g)) as a function of intensity of the applied magnetic field at 7 khz in the range of mt for iron oxide nanocubes suspension. 4
5 T ( C) W/cm W/cm 2.8 W/cm 2.3 W/cm time (min) Figure S7. Heat generation by iron oxide nanocubes in suspension as a function of power intensity of 88 nm-laser. Temperature increase in the LASER mode at [Fe] = 12 mm, for different NIR-laser powers (.3,.8, 1.5, 2.5 W/cm²). T ( C) MHT Fe 3 O 4 NCs.8 W/cm 2 CoFe 2 O 4 NPs Fe 2 O 3 NPs.3 W/cm 2 LASER DUAL LASER DUAL Figure S8. Heat generation by different magnetic nanomaterials in suspension subjected to the three treatments. Temperature increase after 1 s for three magnetic materials (2 nm-iron oxide nanocubes, 9 nm-cobalt ferrite nanoparticles and 9 nm-iron oxide nanoparticles) after applying the different treatments (MHT at 9 khz, 25 mt, LASER at.3 and.8 W/cm 2 and DUAL). The iron concentration was [Fe] = 25 mm in all samples. For all three magnetic nanomaterials, the magnetic and laser heating are cumulative. Because of their excellent magnetic heating performance, iron oxide nanocubes are the most efficient nano-heaters in the DUAL-mode. 5
6 8 6 SKOV-3 g Fe/cell Fe [mm] Figure S9. Cell capture profile (in SKOV-3 cells) of nanocubes as a function of the extracellular iron concentration. At [Fe] =.2 mm, the maximum cellular uptake was reached. 6
7 Figure S1. Therapeutic efficiency in vitro. Detection of the expression of apoptosis-related proteins in cell lysates after the different treatments (MHT, LASER and DUAL (MHT + LASER) for 1 s, and DUAL (MHT + LASER) for 6 s) normalized to the control condition. The list includes caspases activators & regulators, Bcl-2 proteins involved in the intrinsic pathway of caspase activation, proteins reflecting a mitochondrial damage, DNA damage, inhibitors of apoptosis and regulators, heat shock proteins and death receptors involved in the extrinsic pathway of caspase activation. 7
8 Normalized tumor growth Control DUAL non-injected Days Figure S11. In vivo therapy control tumors. Average tumor growth in non-injected mice non-treated (Control) and submitted to the DUAL protocol (DUAL non-injected) during the 8 days following the 3 days of treatment (groups of 6 and 4 mice, respectively). Figure S12. Transmission electron micrographs of tumor tissue. Transmission electron micrographs comparing tumoral stromata of a control and a regressed tumor. (A) General view showing large zones containing tightly bound and organized collagen fibers in the untreated tumor. (B) General view showing large zones of denatured nanocube-loaded collagen zones and occasional scattered, poorly organized collagen fibers bundles that are not loaded with nanocubes (scale bars 5 micrometers). 8
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