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 journalArticle

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.

Original languageEnglish (US)
Pages (from-to)34235-34243
Number of pages9
JournalRSC Advances
Volume9
Issue number59
DOIs
StatePublished - Jan 1 2019

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Micelles
Polyethylene glycols
Photons
Integrins
Cells
Cytotoxicity
Calcium
Fluorescence
Ions
Imaging techniques
polycaprolactone
benzo-1,2,3-thiadiazole
Calcium Chloride
Calcium chloride
Fluorophores
Dynamic light scattering
Bromides
Cell culture
Peptides
Block copolymers

ASJC Scopus subject areas

  • Chemistry(all)
  • Chemical Engineering(all)

Cite this

@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",
year = "2019",
month = "1",
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doi = "10.1039/c9ra06694b",
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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

PY - 2019/1/1

Y1 - 2019/1/1

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.

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