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Polypyrrole-Methylene Blue Nanoparticles as a Single Multifunctional Nanoplatform for Near-Infrared Photo-induced Therapy and Photoacoustic

Imaging

by

Thi Tuong Vy Phan,a,c Subramaniyan Bharathiraja,a Van Tu Nguyen,a,c Madhappan Santha Moorthy,a

Panchanathan Manivasagan,a Kang Dae Lee,d and Junghwan Oh,a,b,*

aMarine-Integrated Bionics Research Center, Pukyong National University, Busan 48513, Republic of

Korea.bDepartment of Biomedical Engineering and Center for Marine-Integrated Biotechnology (BK21 Plus),

Pukyong National University, Busan 48513, Republic of Korea.cInterdisciplinary Program of Biomedical Mechanical & Electrical Engineering, Pukyong National

University, Busan 48513, Republic of Korea.dDepartment of Otolaryngology – Head and Neck Surgery, Kosin University College of Medicine, Busan

48513, Republic of Korea

*The corresponding author

Email: jungoh@pknu.ac.kr

Fax: (82)51-629-5779

Office phone: (82)51-629-5771

Electronic Supplementary Material (ESI) for RSC Advances.This journal is © The Royal Society of Chemistry 2017

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Figure S1: (a) The standard curve of MB, and (b) The plot of the loading amount of MB on the PPy NPs.

y = 0.007x - 0.0007(R² = 1)

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0 20 40 60 80 100

Abso

rptio

n (a

.u.)

Concentration (µg/ml)

Figure S2: The standard curve of PPy-MB NPs (based on 808 nm peak absorption).

y = 0.2038x + 0.1163R² = 0.9889

0

0.5

1

1.5

2

2.5

3

0 2 4 6 8 10 12 14

Abso

rptio

n (a

.u)

Concentration (µg/ml)

(a) (b)

0

0.5

1

1.5

2

2.5

3

0 0.5 1 1.5 2 2.5 3

Load

ing

ratio

(%)

(Mas

s of

MB/

Mas

s of

PPy

-MB

NPs

)*10

0%)

A mount of feeding MB (mg)

3

Figure S3: (a) Zeta potential of PPy, PPy-SDS and PPy-MB NPs, (b) Zeta potentials are recorded for PPy-MB NPs over 12 days.

Figure S4: (a) The UV-Vis-NIR absorption spectra of PPy-MB NPs at the first day and the 60th day of long-term storage test. (b) Decay profiles of the DPBF consumed in the presence of PPy-MB NPs (at 50 µg/ml concentration) with 808 nm laser at 0.5 W/cm2 at the first day and the 60th day. (c) The photos of PPy-MB NPs in different media after 20 days storage. (d) The particles size of PPy-MB NPs at the first

day and the 60th day of long-term storage test.

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

500 600 700 800 900

Abso

rptio

n (a

.u.)

Wavelength (nm)

day 0day 60

(a)

60

65

70

75

80

85

90

95

100

105

0 5 10 15 20 25

DPBF

com

sum

e (%

)

Time (min)

day 0day 60

(b)

0

10

20

30

40

50

60

day 0 day 60

SIze

(nm

)

(d)(c)

-35

-30

-25

-20

-15

-10

-5

0

0 3 6 9 12

Zeta

pot

entia

l (m

V)

Time (Days)

(b)

+14.77

-37.71

-28.47

-60-50-40-30-20-100102030405060

-45

-35

-25

-15

-5

5

15

25

35

45

Size

(nm

)

Zeta

pot

eint

ial (

mV)

(a)

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Figure S5: The UV-Vis-NIR absorption spectra of PPy-MB NPs with DPBF (diluted in DMSO) under 808 nm laser during 0-25 minutes.

Figure S6: FTIR of PPy-MB NPs before and after 808 nm laser (1 W/cm2) irradiation for 30 minutes.

5001000150020002500300035004000

Tran

smitt

ance

(%)

Wavelength (cm-1)

Before irradiation

After irradiation

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Figure S7: The cell viability (with MTT assay) of (a) MDA-MB-231 cells and (b) NHDF-Neo cells

incubated with PPy-MB NPs with different concentrations for 24, 48, and 72 hours.

Figure S8: Fluorescence images of MDA-MB-231 cells incubated with PPy-MB NPs with different concentrations for 24 hours. (a) Control without nanoparticles (b) Incubated with 50 µg/ml PPy-MB NPs,

(c) Incubated with 100 µg/ml PPy-MB NPs, and (d) Incubated with 200 µg/ml PPy-MB NPs.

0

10

20

30

40

50

60

70

80

90

100

110

0 50 100 200 300 400 500 600 700 800 900 1000

Cel

l via

bilit

y (%

)

Concentration of PPy-MB (µg/ml)

24h 48h 72h

0

10

20

30

40

50

60

70

80

90

100

110

0 50 100 200 300 400 500 600 700 800 900 1000

Cel

l via

bilit

y (%

)

Concentration of PPy-MB (µg/ml)

24h 48h 72 h

(a) (b)

(c) (d)

200 µm 200 µm

200 µm200 µm

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Figure S9: The cell viability (with MTT assay) of MDA-MB-231cells treated with PPy NPs, and PPy-MB NPs at various concentrations. The cells were irradiated under 808 nm laser for 6 minutes at 0.5

W/cm2.

0

20

40

60

80

100

120

0 10 30 50

Cel

l via

bilit

y (%

)

Concentration (µg/ml)

PPy+Laser PPy-MB+Laser

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Figure S10: Experimental setup of photoacoustic microscopy (PAM) laser diode on PPy-MB NPs injected samples.

63 µg/ml 125 µg/ml 250 µg/ml 500 µg/ml

0.60.40.2

(b)

(c)

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

600 700 800 900

Abs

orpt

ion

(a.u

.)

Wavelength (nm)

PPy-MBPPy

905 nm

(a)

PPy

PPy-

MB

Figure S11: (a) The UV-Vis absorption of PPy and PPy-MB NPs (the amount of PPy is 50 µg/mL), (b) and (c) PA responses of PPy NPs and PPy-MB NPs at various concentrations.

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y = 0.0008x + 0.0603(R² = 0.9149)

0

0.1

0.2

0.3

0.4

0.5

0 100 200 300 400 500

PA A

mpl

itude

(a.u

.)

Concentration (µg/ml)

y = 0.0007x + 0.0543(R² = 0.9176)

0

0.1

0.2

0.3

0.4

0.5

PA A

mpl

itude

(a.u

.)

Concentration (µg/ml)

(a) (b)

Figure S12: The standard curve of PA amplitude using (a) PPy NPs and (b) PPy-MB NPs.