Size-Modulable Nanoprobe for High-Performance. Ultrasound Imaging and Drug Delivery against

Transcription

1 Size-Modulable Nanoprobe for High-Performance Ultrasound Imaging and Drug Delivery against Cancer Lu Zhang,,,#, Tinghui Yin,#, Bo Li, Rongqin Zheng*,, Chen Qiu, Kit S. Lam*,, Qi Zhang, Xintao Shuai*,, Guangdong Provincial Key Lab of Liver Disease and Department of Medical Ultrasonic, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou , China Department of Biochemistry and Molecular Medicine, UC Davis Comprehensive Cancer Center, University of California Davis, Sacramento, California 95817, USA PCFM Lab of Ministry of Education, School of Materials Science and Engineering, Sun Yat-sen University, Guangzhou , China # These authors contributed equally to this work. Corresponding Authors Xintao Shuai: shuaixt@mail.sysu.edu.cn. Kit S. Lam: kslam@ucdavis.edu. Rongqin Zheng: zhengrq@mail.sysu.edu.cn. 1

2 Supporting Figures Figure S1. Synthetic approach of ph sensitive diblock copolymer mpeg-pasp(dea-co-his-co-dip). 2

3 Figure S2. 1 H NMR spectra of mpeg-pasp(dea-co-his-co-dip) and mpeg-pbla in DMSO-d 6. The DP of PAsp(DIP), PAsp(DEA) and PAsp(His) were calculated to be 50, 20 and 10 by calculating the peak intensity ratio of the terminal CH 3 group of DIP group at 0.95 ppm, the terminal CH 3 group of DEA group at 1.00 ppm and the double bond group of His group at 6.80 ppm, compared to the CH 2 group of PEG at 3.62 ppm. 3

4 Figure S3. FTIR spectra of mpeg-pasp(dea-co-his-co-dip) and mpeg-pbla. After aminolysis reaction, the characteristic peak at 1734 cm -1 of ester (s, v C=O, ester ) disappeared and the characteristic peak at 1658 cm -1 of amide (s, v C=O, amide ) was obviously intensified. Figure S4. GPC traces of mpeg-pasp(dea-co-his-co-dip) and mpeg-pbla with DMF as an eluent. 4

5 Figure S5. Particle sizes of DOX-PPEHD and PFP/PFB/DOX-PPEHD measured by DLS at different conditions. Data were shown as mean ± SD (n = 3). 5

6 Figure S6. Serum stability of PFP/PFB/DOX-PPEHD nanoprobe in (a) PBS solution of ph 7.4 with/without 10% FBS and (b) PBS solution and RPMI-1640 culture medium with 10% fetal bovine serum of ph 6.8. (c) Serum stability of PFP/PFB/DiR-PPEHD nanoprobe in PBS solution of ph 7.4 with 10% FBS and ph 6.8 culture medium (incubation temperature: 37 C, HCl (0.1 N) was used to adjust the ph of culture medium, data were mean ± SD, n = 3). 6

7 Figure S7. Transmission electron microscopy images of (a) DOX-PPEHD nanoprobes and (b) PFP/PFB/DOX-PPEHD nanoprobes at ph 7.4 under room temperature, PFP/PFB/DOX-PPEHD at ph 6.8 under room temperature (c) and under 45 C (d), PFP/PFB/DOX-PPEHD at ph 6.8 plus LFUS under room temperature (e) and under 45 C (f). LFUS if applied: 1 MHz, 50% duty cycle, acoustic pressure 2.0 MPa, 5 min. All samples were stained with uranyl acetate. 7

8 Figure S8. (a) Variation of DOX fluorescence intensity of PFP/PFB/DOX-PPEHD nanoprobe aqueous solution at ph 7.4 (nanoprobe concentration: 0.5 mg ml -1 ). (b) DOX fluorescence intensity changes of PFP/PFB/DOX-PPEHD solution after different LFUS irradiation time at ph 6.8 plus 45 C (nanoprobe concentration: 0.5 mg ml -1 ). (c) DOX fluorescence intensity changes of PFP/PFB/DOX-PPEHD solution after different LFUS irradiation times at ph 6.8 (each time 5 min, nanoprobe concentration: 0.5 mg ml -1 ). All measurements were conducted 2 h after condition adjustment. (d) In vitro quantitative DOX release from the PFP/PFB/DOX-PPEHD aqueous solution (nanoprobe concentration: 1 mg ml -1, data were mean ± SD of three parallel samples). LFUS if applied: 1 MHz, 50% duty cycle, acoustic pressure 2.0 MPa. 8

9 Figure S9. The standard curves of (a) PFP and (b) PFB detected using a gas chromatography mass spectrometry analysis system (GC-MS, Trace GC 2000-DSP). 9

10 Figure S10. In vitro power Doppler imaging of the nanoprobes DOX-PPEHD and PFP/PFB/DOX-PPEHD at different conditions. Nanoprobe concentration: 1.0 mg ml -1. Power Doppler imaging: 15L8-w broadband high-frequency linear transducer, frequency of 10 MHz and a transmitted power of 18 db. 10

11 Figure S11. (a) Variation of DiR fluorescence intensity of PFP/PFB/DiR-PPEHD nanoprobe aqueous solution at different ph. (b) Serum stability of DiR fluorescence spectrum in PFP/PFB/DiR-PPEHD nanoprobe in PBS solution of ph 7.4 with 10% FBS at different time (incubation temperature: 37 C). 11

12 Figure S12. (a) In vivo DiR fluorescence imaging showing tumor accumulation of PFP/PFB/DiR-PPEHD vesicle and DiR-PPEHD vesicle at different time points after tail vein injection into nude mice bearing C6 glioma xenograft. (b) In vivo DiR fluorescence imaging showing tumor non-targeting of the PFP/PFB/DiR-MBs at different time points after tail vein injection into nude mice. (c) Ex vivo DiR fluorescence imaging of the organs and tumors excised at 48 h post-injection time from the same animal. From the left to right of each figure in c show the heart, liver, spleen, lung, kidney and tumor. (DiR dose: 4 mg per kg body weight). 12

13 Figure S13. Tumor-focusing power Doppler US imaging of animals after tail vein injection of PFP/PFB/DOX-PPEHD nanoprobe at 37 C and 45 C. Power Doppler mode: a broadband ML6-15D high-frequency linear transducer, frequency of 7.5 MHz and transmitted power of 18 db. The squares and arrows indicated the imaging zone highlighting tumor. DiR dose in (a): 4 mg per kg body weight, DOX dose in (b): 2.5 mg per kg body weight. 13

14 Figure S14. Body weight curves of the experimental animals receiving different therapeutic formulations via tail vein at an interval of 3 days (n=6, dose per injection: 2.5 mg DOX per kg body weight, LFUS if applied: 1 MHz, 50% duty cycle, acoustic pressure 2.0 MPa, 3 times, 5 min each. Data were mean ± SD of three parallel samples). 14

15 Table S1. Particle size of PFP/PFB/DOX-PPEHD (A) and PFB/DOX-PPEHD (B) at ph 6.8 under different temperatures. All measurements were conducted 30 min after condition adjustment. Data were shown as mean ± SD (n = 3). Temperature ( C) Size of A (nm) 426±16 437±22 485±19 509±18 432±34 362±47 359±44 Size of B (nm) 415±20 423±18 419±22 426±24 441±21 466±26 498±29 15