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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

2

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

8

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.