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KOREAN CHEMICAL SOCIETY
immobile, to allow growing of the seeds in the solution.
Mostly, the solution color was gradually changed within a
few minutes. After a few hours, the GNPs were produced
and washed with water several times followed by a centrifu-
gation at 10 000 rpm for 10 min for several times to remove
free PSs to obtain GNPs as a greenish-black solid.
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Cell Culture and Photo-Irradiation. From the cell line
bank of the Seoul National University Cancer Research Center
A549 (human lung carcinoma) cells were obtained. The cells
were grown in a media solution contains RPMI-1640 (Sigma-
Aldrich, Munich, Germany) with 10% fetal bovine serum and
ꢀ
1
% penicillin at 37 C in a humidified atmosphere with 5%
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CO in air. In the procedures, phosphate-buffered saline (PBS,
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Sigma-Aldrich), trypsin-ethylene diamine tetraacetic acid
ꢀ
(
EDTA) solution, an incubator (37 C, 5% CO ), SynergyHT
2
fluorescence multi-detection reader (BioTek, Winooski, VT,
USA), and optical microscope (model CK40-32 PH; Olympus,
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8, 2350.
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. E. S. Kang, T. H. Lee, Y. Liu, K.-H. Han, W. K. Lee,
Tokyo, Japan) were used. PDT was performed using a LED
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(
735–785 nm, total light dose 2 J/cm for 15 min).
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0. J.-E. Chang, Y. Liu, T. H. Lee, W. K. Lee, I. Yoon, K. Kim,
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MTT Assay and Cell Viability. Cells (1 × 10, 100 μL)
were placed in a 96-well microplate. A 24 h later, the
medium was removed and the plates were washed with
PBS for two times. Mixed medium (100 μL) of each PS
and GNPs were added to the each well. A 24 h later, the
medium was removed and the plates washed with PBS for
three times, followed by adding fresh medium (100 μL).
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After irradiation (2 J/cm , 15 min) of the plates, the MTT
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assay was performed to evaluate PDT activity. In each well
MTT solution was added and incubated for 1 h, and mea-
sured absorbance at 450 nm using a fluorescence multi-
detection reader. The plates were photo-irradiated followed
by 12 and 24 h incubations. Each group consisted of three
wells for three replicate experiments.
1
1
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(1 × 10 cells) were plated on confocal dishes and incubated.
A 24 h later, the medium was removed and dishes were
washed with PBS for three times. Then each PS and GNPs
2
2
(1 μM) were added to the dishes. A 24 h later, the dishes were
washed with PBS for three times. After the fixation solution
was added (10 min), followed by washing with PBS and
staining with diamidino-2-phenylindole (DAPI, 200 nM), cell
images were acquired using CLSM by an LSM 510 META
microscope (Carl Zeiss,Oberkochen, Germany).
Acknowledgments. This research was supported by the
National Research Foundation (NRF) of Korea grant funded
by the Korea government (NRF-2017R1A2B4010615).
2
3. Y.-H. Chen, C.-Y. Tsai, P.-Y. Huang, M.-Y. Chang,
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Supporting Information. Additional supporting informa-
tion is available in the online version of this article.
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Bull. Korean Chem. Soc. 2020, Vol. 41, 230–233
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