Permeation of PEO-PBLA-FITC polymeric micelles in aortic endothelial cells

Jiahorng Liaw, Takao Aoyagi, Kazunori Kataoka, Yasuhisa Sakurai, Teruo Okano

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Purpose. To determine aortic endothelial cells permeation ability and mechanisms of the aqueous block copolymeric micelles, poly(ethylene oxide)- poly (β-benzyl L-aspartate) (PEO-PBLA) chemically conjugated with fluorescein isothiocyanate (FITC) by transport study and confocal laser scanning microscopy. Methods. The block copolymers' PEO-PBLA-FITC was first synthesized and characterized by gel permeation chromatography (GPC) reflect index, UV, fluorescence detectors, and critical micelles concentrations (CMC), and atomic force microscopy (AFM). Permeation ability and mechanisms of polymeric micelles in aortic endothelial cells were evaluated by incubating with NaF, NaN3, wortmannin, cytochalasin B inhibitors, at 20°C, and under reverse conditions. FITC and latex particles (40 nm) were also used for comparison of transport ability. The extent of localization of uptake polymeric micelles was established by confocal laser scanning microscopy. Results. The size of the aqueous PEO-PBLA-FITC polymeric micelles was detected at around 56 nm with unimodal distribution by AFM. The CMC test revealed the fluorescence intensity increased to around 0.01 ~ 0.05 mg/ml. NaF, NAN3, wortmannin, cytochalasin B, 20°C, and reverse experiments inhibited the absorption of polymeric micelles through aortic endothelial cells with apparent permeability coefficients (P) of 18.07 ± 1.03 to 12.98 ± 0.93, 11.31 ± 0.77, 12.44 ± 1.23, 6.40 ± 0.23, 11.11 ± 0.46, and 10.22 ± 1.09 x 10-7 cm/sec, respectively. Also, the permeation of FITC and latex on aortic endothelial cells was 70.02 ± 4,71, and 2.05 ± 0.41 x 10-7 cm/sec, respectively. Confocal laser microscopy showed that fluorescent compounds were distributed in the intracellular cytoplasm and nucleus. Conclusions. PEO-PBLA-FITC copolymeric micelles in an aqueous system were transported by energy-dependent endocytosis with 18.07 x 10-7 cm/sec penetrated range and were localized on intracellular and nucleus endothelial cells.

Original languageEnglish
Pages (from-to)213-220
Number of pages8
JournalPharmaceutical Research
Volume16
Issue number2
DOIs
Publication statusPublished - 1999

Fingerprint

Ethylene Oxide
Endothelial cells
Micelles
Polyethylene oxides
Fluorescein
Aspartic Acid
Permeation
Endothelial Cells
Confocal Microscopy
Microscopic examination
Cytochalasin B
Critical micelle concentration
Latex
Lasers
Atomic Force Microscopy
Atomic force microscopy
Fluorescence
Scanning
Sodium Azide
Hydraulic conductivity

Keywords

  • Endocytosis
  • Endothelial
  • FITC
  • Polymeric micelles

ASJC Scopus subject areas

  • Chemistry(all)
  • Pharmaceutical Science
  • Pharmacology

Cite this

Permeation of PEO-PBLA-FITC polymeric micelles in aortic endothelial cells. / Liaw, Jiahorng; Aoyagi, Takao; Kataoka, Kazunori; Sakurai, Yasuhisa; Okano, Teruo.

In: Pharmaceutical Research, Vol. 16, No. 2, 1999, p. 213-220.

Research output: Contribution to journalArticle

Liaw, Jiahorng ; Aoyagi, Takao ; Kataoka, Kazunori ; Sakurai, Yasuhisa ; Okano, Teruo. / Permeation of PEO-PBLA-FITC polymeric micelles in aortic endothelial cells. In: Pharmaceutical Research. 1999 ; Vol. 16, No. 2. pp. 213-220.
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AU - Sakurai, Yasuhisa

AU - Okano, Teruo

PY - 1999

Y1 - 1999

N2 - Purpose. To determine aortic endothelial cells permeation ability and mechanisms of the aqueous block copolymeric micelles, poly(ethylene oxide)- poly (β-benzyl L-aspartate) (PEO-PBLA) chemically conjugated with fluorescein isothiocyanate (FITC) by transport study and confocal laser scanning microscopy. Methods. The block copolymers' PEO-PBLA-FITC was first synthesized and characterized by gel permeation chromatography (GPC) reflect index, UV, fluorescence detectors, and critical micelles concentrations (CMC), and atomic force microscopy (AFM). Permeation ability and mechanisms of polymeric micelles in aortic endothelial cells were evaluated by incubating with NaF, NaN3, wortmannin, cytochalasin B inhibitors, at 20°C, and under reverse conditions. FITC and latex particles (40 nm) were also used for comparison of transport ability. The extent of localization of uptake polymeric micelles was established by confocal laser scanning microscopy. Results. The size of the aqueous PEO-PBLA-FITC polymeric micelles was detected at around 56 nm with unimodal distribution by AFM. The CMC test revealed the fluorescence intensity increased to around 0.01 ~ 0.05 mg/ml. NaF, NAN3, wortmannin, cytochalasin B, 20°C, and reverse experiments inhibited the absorption of polymeric micelles through aortic endothelial cells with apparent permeability coefficients (P) of 18.07 ± 1.03 to 12.98 ± 0.93, 11.31 ± 0.77, 12.44 ± 1.23, 6.40 ± 0.23, 11.11 ± 0.46, and 10.22 ± 1.09 x 10-7 cm/sec, respectively. Also, the permeation of FITC and latex on aortic endothelial cells was 70.02 ± 4,71, and 2.05 ± 0.41 x 10-7 cm/sec, respectively. Confocal laser microscopy showed that fluorescent compounds were distributed in the intracellular cytoplasm and nucleus. Conclusions. PEO-PBLA-FITC copolymeric micelles in an aqueous system were transported by energy-dependent endocytosis with 18.07 x 10-7 cm/sec penetrated range and were localized on intracellular and nucleus endothelial cells.

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KW - Endothelial

KW - FITC

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