Electronic Supplementary Information (ESI) For pH … · pH-Responsive Mitoxantrone (MX) Delivery Using Mesoporous Silica Nanoparticles (MSN) 5 ... The nitrogen adsorption/desorption

Electronic Supplementary Information (ESI) For

pH-Responsive Mitoxantrone (MX) Delivery Using Mesoporous Silica Nanoparticles (MSN)

5

Yanhang Ma,a†Lin Zhou,b† Haoquan Zheng,a Lei Xing,a Chenguang Li,a Jinghao Cuib* and Shunai Chea*

a School of Chemistry and Chemical Engineering, State Key Laboratory of Metal Matrix Composites, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, P. R. China. Fax: +86 21 5474 1297; Tel: +86

21 5474 2852; E-mail: [email protected] 10

b Department of Pharmaceutics, College of Pharmaceutical Science, Soochow University, 199 Renai Road,

Suzhou 215123,China. Fax: X+86–512–6588–2077; E-mail: [email protected]

1. Material

Cetyltrimethylammonium bromide (CTAB; SCRC, China), tetraethylorthosilicate (TEOS; TCI, 15

Japan), Mitoxantrone (MX). All materials were used as purchased without further purification.

2. Methods

2.1. Synthesis of MCM-41-Type MSN

Typically, 1 g CTAB was first dissolved in the mixture of 480 ml H2O and 7 ml NaOH (1.0 M). The 20

temperature of the solution was adjusted to 80 °C. Then 5.0 ml TEOS was added dropwise to the solution, followed by stirring for 2 h at 80 °C to give rise to white precipitation. The solid product was filtered, washed with deionized water and dried at 100 °C overnight. To remove the surfactant, the as-synthesized materials were extracted in the ethanolic solution of 1 M HCl.

2.2. Synthesis of methylate- and mercapto- functionalized MCM-41 25

The methylate group and mercapto group functionalized MCM-41 nanoparticles were synthesized by post grafting method. Typically, 0.5 g MCM-41 nanoparticles after removal of surfactant were suspended in 20 mL toluene. And then 0.42 mM Methyltriethoxysilane or 3-Mercaptopropyltrimethoxysilane was added dropwise under stirring, followed by refluxing for 12 h.

2.3. Synthesis of carboxyl group functionalized MSN 30

The carboxyl group functionalized mesoporous nanoparticles were synthesized by co-condensation method. 0.14 g C18-3-1 was dissolved in the mixture of 135 mL deionized water and 5.8 g ethanol at 80 °C. And then 0.174 g CES and 0.78 g TEOS were added simultaneously, followed by stirring for 1 h at 80 °C. The obtained solution was aged at 80 °C for 2 days. To removal the surfactant in the nanoparticles, the synthesized materials were extracted with the mixture of 90 mL THF and 10 mL 35% 35

HCl.

2.4. Characterizations

Powder X–ray diffraction (XRD) patterns were recorded on a Rigaku X–ray diffractometer D/MAX–2200/PC equipped with Cu Kα radiation (40 kV, 20 mA) at a rate of 1.0°/min over the range of 1–6° (2θ). The morphology of MCM-41 nanoparticles was observed with scanning electron 40

microscope (SEM, JEOL JSM–7401F) with an accelerating voltage of 1.0 kV. High–resolution transmission electron microscopy (HRTEM) images were taken with a JEOL JEM–3010 microscope operating at 300 kV. The nitrogen adsorption/desorption isotherms were measured at –196 °C with a Quantachrome Nova 4200E porosimeter. The surface area was calculated by the Brunauer–Emmett–Teller (BET). The pore size distribution was calculated by Barrett–Joyner–Halenda (BJH) method 45

according to the adsorption branch of the isotherm. The concentration of anti-cancer drugs (MX) in

Electronic Supplementary Material (ESI) for Journal of Materials ChemistryThis journal is © The Royal Society of Chemistry 2011

solution was measured by a UNICO UV–4802 UV–vis double beam spectrophotometer. The zeta-potential of the nanoparticles was measured in a Malvern ZS90 zeta potential analyzer.

2.5. Loading of MX in the mesopores of the MSN

The mitoxantrone was first prepared as a solution with the concentration of 200 µg/ml in ethanol. In a typical condition, 0.1 g MCM-41 nanoparticles were added into 10 ml MX solution with stirring for 5

24 h. The materials were obtained by centrifugation and then would be washed for three times with PBS of pH 7.4.

2.6. Release of MX from the MSN in PBS solution

In a typical release experiment, about 25 mg of the MCM-41 materials loading with MX was suspended by vibration in 30.0 ml of PBS solution with pH 4.0-7.4 at 37 °C. In the case of sampling, 2 10

ml homogenous solution were withdrew to centrifuged, followed by being measured with UV-vis spectrophotometer.

2.7. In-vitro cell assay

Cells were seeded in a 96-well plate at a seeding density of 5,000 per well in 100µL of RPMI 1640 medium with 10% FBS and 1% penicillin and streptomycin. After the cells were cultured at 37℃ for 15

24h, the growth medium was removed and fresh growth medium containing the predetermined amount of MX-loaded nanoparticles was added. After a 24h incubation, cells were washed three times with 100µL of PBS, and then 100µL of RPMI 1640 medium with 10% FBS was added. Cytotoxicity was assessed using MTT to measure the viability of the cells.10µL of MTT solution (5mg/mL) was added to each well. The plates were incubated for an additional 4h and then 100µL of 10%HCl-SDS were 20

added to dissolve the MTT formazan crystals. After the plates were cultured overnight, the absorbance of each well was measured at 570 nm in a microplate reader.

2.8. Fluorescent experiments

Cells were seeded in a 6 well plate at a seeding density of 2*105 per well in 2 mL of RPMI 1640 medium with 10% FBS and 1% penicillin and streptomycin. After the cells were cultured at 37 °C for 25

24 h with the concentration of CO2 of 5%, the growth medium was removed and fresh growth medium containing the predetermined amount of Rhodamine-loaded nanoparticles was added. After 24 h incubation, cells were washed three times with 2 mL of PBS and then were observed in a confocal laser microscopy.

30

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Electronic Supplementary Information (ESI) For pH … · pH-Responsive Mitoxantrone (MX) Delivery Using Mesoporous Silica Nanoparticles (MSN) 5 ... The nitrogen adsorption/desorption