CRGD functionalized 2,1,3-benzothiadiazole (BTD)-containing two-photon absorbing red-emitter-conjugated amphiphilic poly(ethylene glycol)-: Block -poly(ϵ-caprolactone) for targeted bioimaging

Shanshan Wu; Fengyu Su; Hansa Y. Magee; Deirdre R. Meldrum; Yanqing Tian

doi:10.1039/c9ra06694b

CRGD functionalized 2,1,3-benzothiadiazole (BTD)-containing two-photon absorbing red-emitter-conjugated amphiphilic poly(ethylene glycol)-: Block -poly(ϵ-caprolactone) for targeted bioimaging

Shanshan Wu, Fengyu Su, Hansa Y. Magee, Deirdre R. Meldrum, Yanqing Tian

Research output: Contribution to journal › Article › peer-review

10 Scopus citations

Abstract

A two-photon absorbing (2PA) red emitter group was chemically conjugated onto amphiphilic poly(ethylene glycol)-block-poly(ϵ-caprolactone) (PEG-b-PCL) copolymers, and further grafted with cyclo(Arg-Gly-Asp) (cRGD) peptide to form micelle 1. Micelle 1 with cRGD targeting groups were used for targeted bioimaging. For comparison, micelle 2 without the cRGD targeting groups were also prepared and investigated. The micelles were characterized using dynamic light scattering (DLS), showing average diameters of around 77 nm. The cRGD targeting group is known to bind specifically with α_vβ₃ integrin in cancer cells. In this study, α_vβ₃ integrin overexpressed human glioblastoma U87MG cell line and α_vβ₃ integrin deficient human cervical cancer HeLa cell line were chosen. Results showed that the cRGD targeting group enhanced the cellular uptake efficiency of the micelles significantly in α_vβ₃ integrin rich U87MG cells. Higher temperature (37 °C versus 4 °C) and calcium ions (with 3 M calcium chloride in the cell culture medium versus no addition of calcium ions) enhanced the cellular uptake efficiency, suggesting that the uptake of the micelles is through the endocytosis pathway in cells. A 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) viability assay was used to evaluate the cytotoxicity of the micelles and no significant cytotoxicity was observed. The BTD-containing two-photon absorbing emitter in the micelles showed a two-photon absorbing cross-section of 236 GM (1 GM = 1 × 10^-50 cm⁴ s per photonper molecule) at 820 nm, which is among the highest values reported for red 2PA emitters. Because of the two-photon absorbing characteristics, micelle 1 was successfully used for two-photon fluorescence imaging targeted to U87MG cells under a two-photon fluorescence microscope. This study is the first report regarding the targeted imaging of a specific cancer cell line (herein, U87MG) using the BTD-conjugated-fluorophore-containing block copolymers.

Original language	English (US)
Pages (from-to)	34235-34243
Number of pages	9
Journal	RSC Advances
Volume	9
Issue number	59
DOIs	https://doi.org/10.1039/c9ra06694b
State	Published - 2019

ASJC Scopus subject areas

General Chemistry
General Chemical Engineering

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@article{0801c136d7254d2c9cbcde2d2f9bdfea,

title = "CRGD functionalized 2,1,3-benzothiadiazole (BTD)-containing two-photon absorbing red-emitter-conjugated amphiphilic poly(ethylene glycol)-: Block -poly(ϵ-caprolactone) for targeted bioimaging",

abstract = "A two-photon absorbing (2PA) red emitter group was chemically conjugated onto amphiphilic poly(ethylene glycol)-block-poly(ϵ-caprolactone) (PEG-b-PCL) copolymers, and further grafted with cyclo(Arg-Gly-Asp) (cRGD) peptide to form micelle 1. Micelle 1 with cRGD targeting groups were used for targeted bioimaging. For comparison, micelle 2 without the cRGD targeting groups were also prepared and investigated. The micelles were characterized using dynamic light scattering (DLS), showing average diameters of around 77 nm. The cRGD targeting group is known to bind specifically with αvβ3 integrin in cancer cells. In this study, αvβ3 integrin overexpressed human glioblastoma U87MG cell line and αvβ3 integrin deficient human cervical cancer HeLa cell line were chosen. Results showed that the cRGD targeting group enhanced the cellular uptake efficiency of the micelles significantly in αvβ3 integrin rich U87MG cells. Higher temperature (37 °C versus 4 °C) and calcium ions (with 3 M calcium chloride in the cell culture medium versus no addition of calcium ions) enhanced the cellular uptake efficiency, suggesting that the uptake of the micelles is through the endocytosis pathway in cells. A 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) viability assay was used to evaluate the cytotoxicity of the micelles and no significant cytotoxicity was observed. The BTD-containing two-photon absorbing emitter in the micelles showed a two-photon absorbing cross-section of 236 GM (1 GM = 1 × 10-50 cm4 s per photonper molecule) at 820 nm, which is among the highest values reported for red 2PA emitters. Because of the two-photon absorbing characteristics, micelle 1 was successfully used for two-photon fluorescence imaging targeted to U87MG cells under a two-photon fluorescence microscope. This study is the first report regarding the targeted imaging of a specific cancer cell line (herein, U87MG) using the BTD-conjugated-fluorophore-containing block copolymers.",

author = "Shanshan Wu and Fengyu Su and Magee, {Hansa Y.} and Meldrum, {Deirdre R.} and Yanqing Tian",

note = "Funding Information: Financial support was provided by the Microscale Life Sciences Center, a NIH Center of Excellence in Genomic Sciences at Arizona State University (Grant 5P50 HG002360, Dr Deirdre Meldrum, PI, Director), National Natural Science Foundation of China (21574061, 21601037 and 21604036), Guangdong Industry Polytechnic (KJ2019-003), and Guangdong Province Engineering Technology Centre for Molecular Probe and Biomedicine Imagining. Authors also would like to thank Dr Yongzhong Li and Dr Weiwen Zhang for discussions. Publisher Copyright: {\textcopyright} 2019 The Royal Society of Chemistry.",

year = "2019",

doi = "10.1039/c9ra06694b",

language = "English (US)",

volume = "9",

pages = "34235--34243",

journal = "RSC Advances",

issn = "2046-2069",

publisher = "Royal Society of Chemistry",

number = "59",

}

TY - JOUR

T1 - CRGD functionalized 2,1,3-benzothiadiazole (BTD)-containing two-photon absorbing red-emitter-conjugated amphiphilic poly(ethylene glycol)-

T2 - Block -poly(ϵ-caprolactone) for targeted bioimaging

AU - Wu, Shanshan

AU - Su, Fengyu

AU - Magee, Hansa Y.

AU - Meldrum, Deirdre R.

AU - Tian, Yanqing

N1 - Funding Information: Financial support was provided by the Microscale Life Sciences Center, a NIH Center of Excellence in Genomic Sciences at Arizona State University (Grant 5P50 HG002360, Dr Deirdre Meldrum, PI, Director), National Natural Science Foundation of China (21574061, 21601037 and 21604036), Guangdong Industry Polytechnic (KJ2019-003), and Guangdong Province Engineering Technology Centre for Molecular Probe and Biomedicine Imagining. Authors also would like to thank Dr Yongzhong Li and Dr Weiwen Zhang for discussions. Publisher Copyright: © 2019 The Royal Society of Chemistry.

PY - 2019

Y1 - 2019

N2 - A two-photon absorbing (2PA) red emitter group was chemically conjugated onto amphiphilic poly(ethylene glycol)-block-poly(ϵ-caprolactone) (PEG-b-PCL) copolymers, and further grafted with cyclo(Arg-Gly-Asp) (cRGD) peptide to form micelle 1. Micelle 1 with cRGD targeting groups were used for targeted bioimaging. For comparison, micelle 2 without the cRGD targeting groups were also prepared and investigated. The micelles were characterized using dynamic light scattering (DLS), showing average diameters of around 77 nm. The cRGD targeting group is known to bind specifically with αvβ3 integrin in cancer cells. In this study, αvβ3 integrin overexpressed human glioblastoma U87MG cell line and αvβ3 integrin deficient human cervical cancer HeLa cell line were chosen. Results showed that the cRGD targeting group enhanced the cellular uptake efficiency of the micelles significantly in αvβ3 integrin rich U87MG cells. Higher temperature (37 °C versus 4 °C) and calcium ions (with 3 M calcium chloride in the cell culture medium versus no addition of calcium ions) enhanced the cellular uptake efficiency, suggesting that the uptake of the micelles is through the endocytosis pathway in cells. A 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) viability assay was used to evaluate the cytotoxicity of the micelles and no significant cytotoxicity was observed. The BTD-containing two-photon absorbing emitter in the micelles showed a two-photon absorbing cross-section of 236 GM (1 GM = 1 × 10-50 cm4 s per photonper molecule) at 820 nm, which is among the highest values reported for red 2PA emitters. Because of the two-photon absorbing characteristics, micelle 1 was successfully used for two-photon fluorescence imaging targeted to U87MG cells under a two-photon fluorescence microscope. This study is the first report regarding the targeted imaging of a specific cancer cell line (herein, U87MG) using the BTD-conjugated-fluorophore-containing block copolymers.

AB - A two-photon absorbing (2PA) red emitter group was chemically conjugated onto amphiphilic poly(ethylene glycol)-block-poly(ϵ-caprolactone) (PEG-b-PCL) copolymers, and further grafted with cyclo(Arg-Gly-Asp) (cRGD) peptide to form micelle 1. Micelle 1 with cRGD targeting groups were used for targeted bioimaging. For comparison, micelle 2 without the cRGD targeting groups were also prepared and investigated. The micelles were characterized using dynamic light scattering (DLS), showing average diameters of around 77 nm. The cRGD targeting group is known to bind specifically with αvβ3 integrin in cancer cells. In this study, αvβ3 integrin overexpressed human glioblastoma U87MG cell line and αvβ3 integrin deficient human cervical cancer HeLa cell line were chosen. Results showed that the cRGD targeting group enhanced the cellular uptake efficiency of the micelles significantly in αvβ3 integrin rich U87MG cells. Higher temperature (37 °C versus 4 °C) and calcium ions (with 3 M calcium chloride in the cell culture medium versus no addition of calcium ions) enhanced the cellular uptake efficiency, suggesting that the uptake of the micelles is through the endocytosis pathway in cells. A 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) viability assay was used to evaluate the cytotoxicity of the micelles and no significant cytotoxicity was observed. The BTD-containing two-photon absorbing emitter in the micelles showed a two-photon absorbing cross-section of 236 GM (1 GM = 1 × 10-50 cm4 s per photonper molecule) at 820 nm, which is among the highest values reported for red 2PA emitters. Because of the two-photon absorbing characteristics, micelle 1 was successfully used for two-photon fluorescence imaging targeted to U87MG cells under a two-photon fluorescence microscope. This study is the first report regarding the targeted imaging of a specific cancer cell line (herein, U87MG) using the BTD-conjugated-fluorophore-containing block copolymers.

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U2 - 10.1039/c9ra06694b

DO - 10.1039/c9ra06694b

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SN - 2046-2069

VL - 9

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EP - 34243

JO - RSC Advances

JF - RSC Advances

IS - 59

ER -

CRGD functionalized 2,1,3-benzothiadiazole (BTD)-containing two-photon absorbing red-emitter-conjugated amphiphilic poly(ethylene glycol)-: Block -poly(ϵ-caprolactone) for targeted bioimaging

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