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Electronic Supplementary Information (ESI) For
pH-Responsive Mitoxantrone (MX) Delivery Using Mesoporous Silica Nanoparticles (MSN)
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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.
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I. SEM
The SEM
II. N2 a
Figure nanopa
The N2 aThe measu(Barrett-Joy
M images o
M image sho
adsorption/
S2. (a) N2 aarticles after
Table
MCM-41
adsorption/dured BET (Byner-Halend
of MCM-41
Figure
ows that the
/desorption
adsorption/dr calcinations
S1. Porous
Surface a
8
a–c Cal
desorption aBrunauer-Eda) average
1 nanoparti
e S1. SEM im
e MSN are s
n results of
desorption iss shown in F
and compos
areaa (m2/g)
834
culated from
analysis of mmett-Tell pore diame
icles
mages of MC
spherical sh
f MCM-41
sotherm andFigure 1.
sitional prop
Pore volu
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m N2 adsorp
the extracteler) surface eter was 2.2
CM-41 nano
hape with th
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perties of MC
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ption/desorpt
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25 nm and th
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CM-41 nano
) Pore s
2
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revealed ty834 m2/g. he pore volu
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oparticles.
sizec (nm)
2.25
ypical type The corresume was 0.
CM-41
IV isothermsponding BJ73 cm3/g.
ms. JH
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II
Fi
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II. Mole
igure S3. (a
The two seigure S3a. A07 nm (Fig
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V. Load
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ystem at dif
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ed when thee on silanolo the loss of
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under variou
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nd (c) pH 4.0
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us pH condi
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from 3 to 8when the pHX.
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8, H
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V. Rel
Figure S5functionalize
Fig. S5a
group functpH 7.4. Fodelivery. Tmolecules. bonding ordeterminedgroups andneeded to fubecause of
ease of MX
. (a) 6% ed MCM-41
shows the tionalized s
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The pH-resr electrostat
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files of MXigh release (as shown hat organiclivery of Mions. The qemical analre 6.0 and reason. Fig. nteraction b
ic group fu
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X in variouspercentagein Fig. S4b
c groups coMX in Fig. S
quantitativelysis. The re6.3 mmol/gS4c reveals
between carb
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pH
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FiM4.5
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afThh Ade15
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20
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VI. The s
igure S6. XRCM-41nano0) for 48h.
The presenfter loading he surface aand 48 h. H
An obvious emonstratesrea after treaeleased at pHfter treated f
stability of
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MCM-41 M
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Table S
MC
Materials
oaded MCM-d in 7.4 for 4d in 4.0 for 4lated from N
k (2θ~2.5°e the drug. Nvolume and at pH 4.0, Mof surface aof mesopo
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the mainteresults showll decrease be released t 676 m2/g,een occupiet be caused e in PBS so
ms and pore 4) for 48h; (
tional prope
various stage
Pore volum(cm3/g)
0.3 0.2 0.538
n data.
enance of mw that the cafter treatedand mesop
, has been d by MX. Tby the colla
olution at pH
size distribuc) materials
erties of
es.
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ution of (a) Ms released in
sizec m) .8 .8 .2
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MX-loaded n PBS (pH
anoparticlesf mesoporesH 7.4 for 24y preservedMX, which
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at at pH 7.4
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VII. Effe
This resu
a favorable VIII. N2
fun
Figure S8functionalize
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Figure
ult showed t drug carrie
adsorptioctionalized
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that the MCer for cancer
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CM-41 nanor therapy.
on results
rption isotheup functiona
cancer cell.
SMMC-7721
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.
cells for MC
how low cyt
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totoxicity an
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distributioncapto group
oparticles.
and could be
and mer
n of (a) cap functionaliz
e employed
capto grou
arboxyl grozed MCM-41
as
up
oup 1.
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Table S2.
All the threunctionalizehe pore size
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X. Uptand Figureprresp To skindincrcomwhi
take efficied MCF-7.
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he high inhib
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ell lines, Rhwn in the rest increases. ncy in A54
SMMC-772
ells. 1, 2 and1000 mg/m
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