External Magnetic Field Enhanced Chemo-Photothermal Combination Tumor. Therapy via Iron Oxide Nanoparticles

Transcription

1 Supporting information External Magnetic Field Enhanced Chemo-Photothermal Combination Tumor Therapy via Iron Oxide Nanoparticles Xiaomeng Guo, Wei Li, Lihua Luo, Zuhua Wang, Qingpo Li, Fenfen Kong, Hanbo Zhang, Jie Yang, Chunqi Zhu, Yongzhong Du, and Jian You* College of Pharmaceutical Sciences, Zhejiang University, 866 Yuhangtang Road, Hangzhou, Zhejiang , P. R. China * Corresponding Author: Jian You, College of Pharmaceutical Sciences, Zhejiang University, 866 Yuhangtang Road, Hangzhou , Zhejiang, China. Office: Fax: youjiandoc@zju.edu.cn. S-1

2 Figure S1. Characterization of Fe 3 O 4-10, Fe 3 O 4-60, Fe 3 O 4-120, Fe 3 O and Fe 3 O nanoparticles. (A) TEM images of the Fe 3 O 4 nanoparticles. Scale bar, 200 nm. (B) The size distribution of different sized nanoparticles. (C) XRD graphs of standard Fe 3 O 4 and the synthesized Fe 3 O 4-10 nanoparticles. (D) Magnetization of Fe 3 O 4-10 nanoparticles. S-2

3 Figure S2. Characterization of DOX loaded magnetic nanoparticles. (A) Storage stability of different sized DOX loaded nanoparticles at 37 C for 10 and 24 h. (B) Plasma stability of DOX loaded nanoparticles after 1, 3, 6, and 24 h incubation at 37 C. (C and D) Release profile of DOX from the five types of DOX loaded nanoparticles at different ph values (7.4 and 5.5). All of the data are represented as the average ± SD. S-3

4 Figure S3. Photos of different sized nanoparticle solutions (in PBS) with the same concentration of Fe 3 O 4 under the external magnetic field over time. S-4

5 Figure S4. Photos of different sized nanoparticle solutions (in solution with the similar viscosity of blood) with the same concentration of Fe 3 O 4 under the external magnetic field over time. S-5

6 Figure S5. Photos of different sized drug loaded nanoparticle solutions (in PBS) with the same concentration of Fe 3 O 4 under the external magnetic field over time. S-6

7 Figure S6. Photos of different sized drug loaded nanoparticle solutions (in solution with the similar viscosity of blood) with the same concentration of Fe 3 O 4 under the external magnetic field over time. S-7

8 Figure S7. Magnetic retention of different sized nanoparticles in simulated blood circulation system over time (flow velocity: artery, cm/s, in PBS). S-8

9 Figure S8. Magnetic retention of different sized nanoparticles in simulated blood circulation system over time (flow velocity: artery, cm/s, in solution with the similar viscosity of blood). S-9

10 Figure S9. Magnetic retention of different sized drug loaded nanoparticles in simulated blood circulation system over time (flow velocity: artery, cm/s, in PBS). S-10

11 Figure S10. Magnetic retention of different sized drug loaded nanoparticles in simulated blood circulation system over time (flow velocity: artery, cm/s, in solution with the similar viscosity of blood). S-11

12 Figure S11. (A) Cytotoxicity of MCF-7 cells at 24 h after treatment with Fe 3 O 4-10, Fe 3 O 4-60, Fe 3 O 4-120, Fe 3 O 4-200, and Fe 3 O nanoparticles. (B) Cytotoxicity of MCF-7 cells at 48 h after treatment with different sized Fe 3 O 4 nanoparticles. (C) Cytotoxicity of MCF-7 cells at 24 h after treatment with Fe 3 O 4-10, Fe 3 O 4-60, Fe 3 O 4-120, Fe 3 O 4-200, and Fe 3 O nanoparticles with the MF. (D) Cytotoxicity of MCF-7 cells at 48 h after treatment with different sized Fe 3 O 4 nanoparticles with the MF. All of the data are represented as the average ± SD. S-12

13 Figure S12. Time dependent cellular uptake of MCF-7 cells incubated with different nanoparticles (at the same concentration) for 1 h, 2 h, 4 h, 12 h, 24 h, and 48 h with or without the magnet. Scale bar, 100 µm. S-13

14 Figure S13. Fluorescence imaging of mice bearing bilateral subcutaneous S180 tumors on opposite flanks after intravenously injection of ICG labeled different sized Fe 3 O 4 nanoparticles. Red line circles tumors with a magnet attached. S-14

15 Figure S14. Confocal images of tumor slices taken from bilateral tumor modal mice injected with different sized Fe 3 O 4 nanoparticles with or without a magnet attached on the tumor. A great number of Fe 3 O 4 nanoparticles with strong DOX fluorescence could be found in the MF attached tumor, whereas in the other tumor without magnet attached, a rather weak DOX fluorescence was observed. S-15

16 Figure S15. Magnetic resonance imaging study of different sized Fe 3 O 4 nanoparticles. (A) T 1 -weighted images of aqueous solutions containing Fe 3 O 4-10, Fe 3 O 4-60, Fe 3 O 4-120, Fe 3 O 4-200, or Fe 3 O nanoparticles at various concentrations. (B) T 2 -weighted images of aqueous solutions containing different sized Fe 3 O 4 nanoparticles at various concentrations. (C) T 1 relaxation rates (R 1 ) of four types of nanoparticle solutions at different Fe concentrations. (D) T 2 relaxation rates (R 2 ) of four types of nanoparticle solutions at different Fe concentrations. (E) T 2 weighted images of MCF-7 cells after incubation with different nanoparticles for 1, 24, and 48 h. (F) R 2 values of MCF-7 cells after incubation with different nanoparticles. All of the data are represented as the average ± SD. S-16

17 Figure S16. In vivo T 2 -weighted MR images of mice after intravenously injection of different sized Fe 3 O 4 nanoparticles. Green and white arrows point to tumors on the left or right side, respectively. The left tumors in the right panel were attached with a magnet during the experiment. S-17

18 Figure S17. T 2 -weighted MR image intensity of mice in Figure S15 after intravenously injection of different sized Fe 3 O 4 nanoparticles. (A) T 2 -weighted MR image intensity of tumors without magnet at 1 h post injection. (B) T 2 -weighted MR image intensity of tumors with magnet at 1 h post injection. (C) T 2 -weighted MR image intensity of tumors without magnet at 2 h post injection. (D) T 2 -weighted MR image intensity of tumors with magnet at 2 h post injection. All of the data are represented as the average ± SD; the data were analyzed by one-way ANOVA (0.01 < *P 0.05; **P 0.01;). S-18

19 Figure S18. (A) The growth curves of tumors was monitored over the course of the study (n = 6). (B) All the S180 tumors (n = 6) were collected and weighted after the mice were sacrificed. (C) The photographs of tumors after 30 day-treatment. All of the data are represented as the average ± SD; the data were analyzed by one-way ANOVA (0.01 < *P 0.05; **P 0.01;). S-19

20 Figure S19. H&E staining of major organs sections obtained from mice models 30 days post nanoparticle implantation following intravenous injection on days 0, 3, 6 and 9. Scale bars are 100 µm. S-20

21 Table S1. Synthesis Parameters of Different-Sized Fe 3 O 4 Nanoparticles Prepared by Solvothermal Method Sample FeCl 3 6H 2 O (g) Na 3 Cit (g) NaOAc (g) PEG (g) EG (ml) Time (h) Fe 3 O Fe 3 O Fe 3 O Fe 3 O Table S2. Stability Study of Fe 3 O 4 Nanoparticles in PBS or Plasma NPs Size (nm), in PBS Size (nm), in plasma 0 h 6 h 24 h 0 h 6 h 24 h Fe 3 O ± ± ± ± ± ± 4.8 Fe 3 O ± ± ± ± ± ± 9.4 Fe 3 O ± ± ± ± ± ± 12.4 Fe 3 O ± ± ± ± ± ± 9.2 Fe 3 O ± ± ± ± ± ± 13.5 Table S3. Stability Study of DOX-loaded Fe 3 O 4 Nanoparticles in PBS or Plasma DOX-loaded Size (nm), in PBS Size (nm), in plasma NPs 0 h 6 h 24 h 0 h 6 h 24 h Fe 3 O ± ± ± ± ± ± 6.3 Fe 3 O ± ± ± ± ± ± 8.4 Fe 3 O ± ± ± ± ± ± 9.5 Fe 3 O ± ± ± ± ± ± 10.3 Fe 3 O ± ± ± ± ± ± 12.5 S-21

22 Table S4. Relaxivities (mm 1 s 1 ) and r 2 /r 1 Ratios of Fe 3 O 4 Nanoparticles Sample Relaxivity value [mm 1 s 1 ] r 1 r 2 r 2 / r 1 Fe 3 O Fe 3 O Fe 3 O Fe 3 O Fe 3 O S-22