biocompatibility cargo carrier with enhanced tumor-homing … · 2014-10-21 · 1 supplementary...
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Supplementary Information
Amphiphilized Poly(ethyleneimine) Nanoparticles: A Versatile Multi-
Cargo Carrier with Enhanced Tumor-Homing Efficiency and
Biocompatibility
Solji Park,a,b,† Keunsoo Jeong,a,c,† Eunjung Lee,a,† Jae Hyuk Lee,b Ji Young Yhee,a Ajay Singh,a,d
Joonseok Koh,d Sangyoup Lee,a Kwangmeyung Kim,a Ick Chan Kwon,a Chong Rae Park,c Jungahn
Kim,b,* Sehoon Kima,*
aCenter for Theragnosis, Korea Institute of Science and Technology, 39-1 Hawolgok-dong,
Seongbuk-gu, Seoul 136-791, KoreabDepartment of Chemistry, Kyung Hee University, Dongdaemoon-gu, Seoul 130-701, KoreacCarbon Nanomaterials Design Laboratory, Global Research Laboratory and Department of
Materials Science and Engineering, Seoul National University, Seoul 151-744, KoreadDepartment of Textile Engineering, Konkuk University, Seoul 143-701, Korea
Table S1. Molecular weights of aPEIs determine by 1H-NMR spectrometry
bPEI aPEI-0 aPEI-6.3 aPEI-25
NC181 - 5.2 5.2 5.2
NPEG1 - - 1.9 7.7
MW (kDa) 1.8 3.1 12.6 41.6
1 Number of conjugated octadecyl or PEG pendants per bPEI chain.
Electronic Supplementary Material (ESI) for Journal of Materials Chemistry B.This journal is © The Royal Society of Chemistry 2014
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1580N-H def.
2918, 2850
3462, 3260
C-H str.
N-H str. 1470C-H def.
1120C-N str.
aPEI-0
aPEI-6.3
Tans
mitt
ance
3445N-H str.O-H str.(adsorbed water)
2888C-H str. (PEG)
1342C-H def. (PEG)
1110C-O-C str. (PEG)
4000 3500 3000 2500 2000 1500 1000
aPEI-25
Wavenumber (cm-1)
3445N-H str.O-H str.(adsorbed water) 2888
C-H str. (PEG)1342
C-H def. (PEG)
1110C-O-C str. (PEG)
Fig. S1 FT-IR spectra of aPEIs.
30 nm 30 nm
(a) (b) 14.7 ± 2.6 nm24.7 ± 6.9 nm
Fig. S2 TEM images of (a) aPEI-0 and (b) aPEI-6.3 NPs. The average particle sizes were indicated.
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Fig. S3 Temporal absorbance change showing the colloidal stability of aPEI NPs (1 mg mL-1 in
deionized water (a) and 10% serum at 37C (b)): (black) aPEI-0, (red) aPEI-6.3, and (blue) aPEI-25.
The agglomerates of NPs formed in the course of time were removed with a syringe filter (pore
diameter 0.2 m) prior to every spectral measurement.
Log C (mM)
I 330
/ I32
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1E-8 1E-7 1E-6 1E-5 1E-4 1E-3 0.01 0.10.97
0.98
0.99
1.00
1.01
1.02
1.03
1.04
Fig. S4 Critical micelle concentration (CMC) was measured based on the partition change of pyrene
between the hydrophobic core and the aqueous environment upon micelle formation. The CMC
value of aPEI NPs was estimated as 6.3 M by monitoring the intensity ratio of the pyrene excitation
bands at 330 and 328 nm (I330/I328) as a function of particle concentration.
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50 μm
50 μm
Ctrl
Rub
rene
Optical Fluorescence
Fig. S5 Optical and fluorescence images of HeLa cells, non-treated (Ctrl) and treated with rubrene
for 12 h of incubation.
Ctrl 12 h 24 h
Opt
ical
Cy5
.5R
ubre
ne
50 μm 50 μm 50 μm
Fig. S6 Optical and fluorescence (Cy5.5 and rubrene channels) images of HeLa cells treated with
Cy5.5-labeled aPEI-25 NPs encapsulating rubrene for 12 and 24 h of incubation.
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Fig. S7 In vitro cytotoxicity of aPEI-25 NPs against HeLa cells, evaluated by the colorimetric MTT
assay. Cells were treated with NPs for 1 and 24 h of incubation and assayed in comparison with
untreated cells (ctrl).
aPE
I-6.3
5 m 1 h 1 d 7 d12 hBefore
aPE
I-0aP
EI-6
.3aP
EI-0
5 m 1 h 1 d 7 d12 hBefore
(a)
(b)
1.4
1.0
0.8
0.2 Fluo
resc
ence
1.4
1.2
1.0
0.8 Fluo
resc
ence
1.6
Fig. S8 In vivo NIRF images showing the biodistribution of Cy5.5-labeled aPEI-0 and aPEI-6.3 NPs
encapsulating IR780, intravenously injected into SCC7 tumor-bearing mice (n = 5). 2-Channel NIRF
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signals were taken for Cy5.5 (a: ex = 675 nm, em = 720 nm) and IR780 (b: ex = 745 nm, em = 800
nm) channels, at the indicated time points before and after tail vein injection of the NPs. Red circles
indicate the tumor location.
(a)
(b)
IR78
0C
y5.5
Fluo
resc
ence
2.0
4.0
6.0
3.5
4.0
4.5
IR78
0C
y5.5
2.0
4.0
6.0
3.2
3.6
4.0
Fluo
resc
ence
IR78
0
4.0
5.0
6.0
Fluo
resc
ence
(c)
Fig. S9 Ex vivo NIRF images showing the biodistribution of free IR780 (a) and IR780 loaded in the
Cy5.5-labeled aPEI NPs (b: aPEI-0 NPs and c: aPEI-6.3 NPs) in SCC7 tumor-bearing mice. The
organs were collected from the mouse bodies at 7 d post-injection. 2-Channel NIRF signals were
taken for Cy5.5 (ex = 675 nm, em = 720 nm) and IR780 (ex = 745 nm, em = 800 nm).
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Pdot aPEI-25 aPEI-25/Pdot-40-30-20-10
0102030
Zeta
pot
entia
l (m
V)
Fig. S10 Zeta potentials of Pdot, aPEI-25 NPs, and Pdot-loaded aPEI-25 NPs (Pdot: aPEI-25 = 1:100
by weight).
0 0.65 6.5 32.5 65
Fig. S11 Gel retardation assay with aPEI-25 NPs/DNA complexes. N/P ratios are indicated for every
lane. The squares in the photo indicate free DNA.
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600 700 800 900
excited at 500 nm excited at 710 nm
FL
Inte
nsity
(a.u
.)
Wavelength (nm)
IR780 in THF
no excitation of IR780at 500 nm
600 700 800 900
indirectly excited IR780 fluorescence by FRET from Pdot
excited at 500 nm
FL In
tens
ity (a
.u.)
Wavelength (nm)
IR780/Pdot dual-loaded aPEI-25 NPs
FRET-quenchedPdot fluorescence
Fig. S12 PL spectra (red: excited at 500 nm, blue; excited at 710 nm) of IR780 solution in THF (left)
and water-dispersed aPEI-25 NPs co-loaded with both Pdot and IR780 (right).
aPE
I-0aP
EI-6
.3aP
EI-2
5
Lung Kidney Liver
200 ㎛ 200 ㎛ 200 ㎛
Fig. S13 Representative histological photomicrographs of hematoxylin (H) and eosin (E) stained
organ sections resected at 7 d post-injection of aPEI NPs.
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aPEI-6.3
aPEI-25
0.4
0.6
0.8
1.0
Che
milu
min
esce
nce
PBS
aPEI-6
.3
aPEI-2
50
1
2
3
4
5
6
7
CL In
tens
ity (a
.u.)
(a) (b)
Fig. S14 (a) Inflammation imaging obtained with a H2O2–responsive chemiluminescence (CL)
nanoprobe in a mouse that was intradermally injected with aPEI-6.3 and aPEI-25 NPs. (b) Generated
CL intensities indicating the degree of inflammation induced by PBS or aPEI NPs.