Supporting Information. Silver Nanoparticle-Gated Mesoporous Silica-Coated Gold Nanorods. Low Premature Release and Multifunctional
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1 Supporting Information Silver Nanoparticle-Gated Mesoporous Silica-Coated Gold Nanorods Low Premature Release and Multifunctional Cancer Theranostic Platform Zhehua Zhang, a, Changhui Liu, a,, Junhui Bai, a Cuichen Wu, d Yue Xiao, a Yinhui Li, a Jing Zheng, a, * Ronghua Yang a,c, * and Weihong Tan a,d a State Key Laoratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, 4182, China; Department of Chemistry and Environmental Engineering, Hunan City University, Yiyang, 413, P. R. China; c School of Chemistry and Biological Engineering, Changsha University of Science and Technology, Changsha, 414, China; d Center for Research at the Bio/Nano Interface, Department of Chemistry and Department of Physiology and Functional Genomics, Shands Cancer Center and UF Genetics Institute, University of Florida, Gainesville, Florida, , USA; These authors contriuted equally to this work *To whom correspondence should e addressed: zhengjing213@hnu.edu.cn; Yangrh@pku.edu.cn Fax: S1
2 EXPERIMENTAL SECTION Instruments. Transmission electron microscopy (TEM) images were otained with a JEOL-31 using an accelerating voltage of 1 kv. Zeta potential experiments and dynamic light scattering (DLS) measurements were performed at 25 o C using a Malvern Zeta Sizer Nanoseries (Nano ZS9). Fourier transform infrared (FT-IR) spectra were otained from a TENSOR 27 spectrometer (Bruker Instruments Inc., Germany). UV-Vis spectra were collected using a Hitachi U-41 spectrophotometer (Kyoto, Japan). All fluorescence measurements were performed on a PTI Fluorescence System (Photo Technology International, Birmingham, NJ). Confocal laser scanning microscopy (CLSM) images were otained on a Fluoview TM FV 1 (Olympus, Japan). MTT assay was otained on a Synergy 2 Multi-Mode Microplate Reader (Bio-Tek, Winooski, TV). Synthesis of AuNRs. CTAB solution (2. ml,.2 M) was mixed with 2. ml of.5 mm HAuCl 4. After which,.24 ml ice-cold NaBH 4 (.1 M) was added with vigorous stirring, the solution was stirring for 2 minutes under 25 ºC until turned to rownish-yellow. Seed solution was aged 2 h efore further use. The growth solution was prepared y mixing CTAB solution (48 ml,.2 M), HAuCl 4 (48 ml, 1. mm) and AgNO 3 (96 μl, 4. mm) together in a 25 ml flask. Upon addition of ascoric acid (672 μl, 8 mm) under stirring, the solution ecame colorless. Finally, AuNRs growth was initiated y adding μl seeds. The solution changed color gradually in 1~2 minutes. Synthesis of AuNR@MS. Mesoporous silica coating on AuNRs was prepared as reported previously with some modifications. Briefly, 3 ml of the as-synthesized AuNRs was centrifuged at 1 rpm for 3 minutes to remove excess CTAB surfactant, then re-dispersed in 14. ml Milli-Q water. Under vigorous stirring, μl (.1 M) NaOH solution was added followed y three injections of 4 μl 2% TEOS-methanol solution at 3 minute intervals. The mixture was reacted for 24 h at room temperature. The as-synthesized AuNR@MS was centrifuged, washed with deionized water and dried under vacuum. Synthesis of AuNR@MS-NH 2. The as-synthesized AuNR@MS otained was dispersed in 14 ml ethanol containing 35 μl APTES, after reaction for 24 h at room temperature, centrifuged at 12 rpm for 3 minutes and the precipitates were washed with ethanol. The otained precipitates was further reacted in 2 ml 1 mg/ml NH 4 NO 3 -ethanol solution under 6 ºC overnight to remove the surfactant template S2
3 CTAB, followed y wash with ethanol and dried under vacuum. Synthesis of AuNR@MS-COOH. 4 mgaunr@ms-nh 2 was added into 12mL DMSO containing 96 mg succinic anhydride and 15 μl triethylamine and stirred at 4 ºC for 24 h. The particle was recovered y centrifugation and washed with ethanol to otain AuNR@MS-COOH. Preparation of AuNR@MS-P1.The conjugation of P1 and AuNR@MS-COOH was ased on the EDC/NHS method. A solution of EDC (187.5 μl, 1 mg/ml) and sulfo-nhs (62.5 μl, 1 mg/ml) in MES uffer (.1 M, ph 6.) was prepared, and 1 mg AuNR@MS-COOH was dispersed into the mixture and incuated at 37 C for 15 minutes to activate the caroxyl group. After adjusting the uffer ph to 7.4 y 1 M NaOH solution, P1 (1 μl, 5 μm) was added, and shaken at 37 C for 6 h. The resultant P1-conjugated AuNR@MS were washed y PB uffer (25 mm, ph 7.4) three times to remove unreacted iomolecules. Effect of Laser Induced Temperature Change. The photothermal property of our nanoparticles were susequently veried through a temperature elevation experiment using a 78 nm laser as the irradiation source. and AuNR@MS were at the concentration of 2 μg/ml, NIR laser irradiated at 3.W/cm 2 for different time. HeLa and L2 cells incuated with AuNR@MS@AgNPs-AS1411 were treated with trypsin, centrifuged at 2 rpm for 5 minutes to remove trypsin and cell culture medium, then re-dispersed in PBS efore temperature measurement. S3
4 4 3 Zeta Potential / mv a c d e f -2-3 Figure S1. Zeta potential measurements for AuNR (a), AuNR@MS (), AuNR@MS-NH 2 (c), AuNR@MS-COOH (d), AuNR@MS-P1 (e) and AuNR@MS@ AgNPs (f). 1 a Si-OH Si-O Transmittance / % OH and NH 2 c C=O N-H Wavenumer / cm -1 Figure S2. FT-IR spectra of the synthesized AuNR@MS (a), AuNR@MS-NH 2 (), and AuNR@MS-COOH (c). S4
5 Fluorscence Intensity / A Fluorscence Intensity / B a Col 11 v Col 16 Col 11 v Col 18 Col 11 v Col 2 Col 11 v Col Wavelength / nm Figure S3. (A) Fluorescence spectra of TMPyP 4 solution (.5 µm) with increasing equivalences of AuNRs (from top to ottom:, 6.3, 15.8, 31, 62, 91, 119 and 147 µm) with an excitation wavelength of 421 nm. (B) Fluorescence spectrum of SOSG efore (a) and after AuNRs addition (). ex = 54 nm. The concentration of TMPyP 4 is.5 µm while the concentration of AuNRs is 147 µm Wavelength / nm 1 c Release/% a Time/h Figure S4. Real-time records of the fluorescence emission at 66 nm, reflecting changes of the AuNR@TMPyP in PBS (a), 1 mm tripeptide (), and 1 mm GSH (c). ex= 421 nm. S5
6 Fluorescence Intensity / P1:Ag + =1:4 P1:Ag + =1:6 P1:Ag + =1:8. a c Figure S5. Relative fluorescence intensity of TMPyP 4 released from AuNR@TMPyP with various molar ratios of Ag + to P1 in PBS (a), in PBS containing 5 % human serum (), and ) +5. mm GSH (c) at room temperature. Cells Cells + AuNR@MS@AgNPs-AS1411-FITC at 37 o C Cells + AuNR@MS@AgNPs-AS1411-FITC at 4 o C Cells + AuNR@MS@AgNPs-AS1411-FITC at 37 o C+trypsin Cells + AuNR@MS@AgNPs-AS1411-FITC at 4 o C+trypsin Figure S6. The internalization of AuNR@MS@AgNPs-AS1411-FITC. Flow cytometry results of AuNR@MS@AgNPs-AS1411-FITC inding with HeLa cells under different conditions. Cells, 5 k/sample. S6
7 Cell viaility / % A Cell viaility / % B Nanoparticle Concentration / Ìg/mL Nanoparticle Concentration / Ìg/mL Figure S7. Cell relative viaility of (A) HeLa and (B) L2 cells after treated with AuNR@MS@AgNPs-AS1411 following y white-light irradiation cell viaility / % Irrdiation time / min Figure S8. Cell relative viaility of HeLa cells treated with 78 nm CW laser irradiation at the dose of 3. W/cm 2 for different duration time. S7
8 12 Cell viaility / % TMPyP 4 Concentration / ÌM Figure S9. Cell relative viaility of HeLa cells upon additions of different concentration of TMPyP 4 following y irradiation 78 nm continuous wave laser at the dose of 3. W/cm 2 for 1 min. Temperature æ a Time/min Figure S1. Temperature-time curves of HeLa cells (a) and L2 cells () upon treated with AuNR@MS@AgNPs-AS1411 followed y 78 nm laser irradiation at the dose of 3. W/cm 2. S8
9 cell viaility (%) without NIR irradition with NIR irrdiation a c Figure S11. Comination of photothermal and photodynamic therapy of L2 cells. cells only (a); a+aunr@ms@agnps-as1411 (); (c). Concentration of AuNR@MS@AgNPs-AS1411 and AuNR@TMPyP were 1 μg/ml. 78 nm laser irradiation at the dose of 3. W/cm 2 for 1 min. S9
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