Perflubron (PFOB) protects cell monolayers against direct in vitro oxidative injury

Alexandre T. Rotta, Björn Gunnarsson, Lynn J. Hernan, Bradley P. Fuhrman, David M. Steinhorn

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Introduction: Oxidative stress is often implicated in the genesis of injury to living tissues, even though intact cells generally possess sophisticated antioxidant systems. PFOB attenuates oxidative lung injury in vivo, at least in part due to a decrease in free radical release by macrophages. We hypothesized that PFOB would also protect a biological system from direct injury by reactive oxygen species. Methods: Rat pulmonary artery endothelial cells (RPAECs) were grown to confluence on optical grade culture dishes using standard technique. Cells were labeled with a permeable intracellular probe containing dihydrochlorofluorescein-diacetate (DCFH-DA), activated by cytoplasmic esterases. The cells were exposed to PFOB or culture medium for 15 min, after which all PFOB was removed by a transfer pipette. The monolayers were washed with medium prior to introduction of a buffer containing H2O2 to induce oxidative stress. Changes in cell fluorescence were measured in a kinetic fashion using a confocal fluorescence microscope. The digital images were analyzed with dedicated software to measure the amount of oxidative stress experienced by each monolayer. Results: Values are means ± SEM.*p < 0.05 vs baseline, 1 p < 0.05 vs PFOB. Conclusions: PFOB attenuates oxidative injury to cell monolayers. We speculate that due to its high spreading coefficient, PFOB may act as a physical barrier against free radicals originated in the aqueous phase. The lipophilic PFOB could potentially alter cell membrane fluidity and confer some protection even after its removal. Supported, in part, by the American Lung Association- NY Affiliate, and by Alliance Pharmaceutical Corporation.

Original languageEnglish (US)
JournalCritical Care Medicine
Volume27
Issue number1 SUPPL.
StatePublished - 1999
Externally publishedYes

Fingerprint

Wounds and Injuries
Oxidative Stress
Free Radicals
Fluorescence
Architectural Accessibility
Membrane Fluidity
In Vitro Techniques
perflubron
Lung Injury
Esterases
Pulmonary Artery
Culture Media
Reactive Oxygen Species
Buffers
Software
Endothelial Cells
Antioxidants
Macrophages
Cell Membrane
Lung

ASJC Scopus subject areas

  • Critical Care and Intensive Care Medicine

Cite this

Rotta, A. T., Gunnarsson, B., Hernan, L. J., Fuhrman, B. P., & Steinhorn, D. M. (1999). Perflubron (PFOB) protects cell monolayers against direct in vitro oxidative injury. Critical Care Medicine, 27(1 SUPPL.).

Perflubron (PFOB) protects cell monolayers against direct in vitro oxidative injury. / Rotta, Alexandre T.; Gunnarsson, Björn; Hernan, Lynn J.; Fuhrman, Bradley P.; Steinhorn, David M.

In: Critical Care Medicine, Vol. 27, No. 1 SUPPL., 1999.

Research output: Contribution to journalArticle

Rotta, AT, Gunnarsson, B, Hernan, LJ, Fuhrman, BP & Steinhorn, DM 1999, 'Perflubron (PFOB) protects cell monolayers against direct in vitro oxidative injury', Critical Care Medicine, vol. 27, no. 1 SUPPL..
Rotta AT, Gunnarsson B, Hernan LJ, Fuhrman BP, Steinhorn DM. Perflubron (PFOB) protects cell monolayers against direct in vitro oxidative injury. Critical Care Medicine. 1999;27(1 SUPPL.).
Rotta, Alexandre T. ; Gunnarsson, Björn ; Hernan, Lynn J. ; Fuhrman, Bradley P. ; Steinhorn, David M. / Perflubron (PFOB) protects cell monolayers against direct in vitro oxidative injury. In: Critical Care Medicine. 1999 ; Vol. 27, No. 1 SUPPL.
@article{069ddd6e4812476a871632fe1a8fa426,
title = "Perflubron (PFOB) protects cell monolayers against direct in vitro oxidative injury",
abstract = "Introduction: Oxidative stress is often implicated in the genesis of injury to living tissues, even though intact cells generally possess sophisticated antioxidant systems. PFOB attenuates oxidative lung injury in vivo, at least in part due to a decrease in free radical release by macrophages. We hypothesized that PFOB would also protect a biological system from direct injury by reactive oxygen species. Methods: Rat pulmonary artery endothelial cells (RPAECs) were grown to confluence on optical grade culture dishes using standard technique. Cells were labeled with a permeable intracellular probe containing dihydrochlorofluorescein-diacetate (DCFH-DA), activated by cytoplasmic esterases. The cells were exposed to PFOB or culture medium for 15 min, after which all PFOB was removed by a transfer pipette. The monolayers were washed with medium prior to introduction of a buffer containing H2O2 to induce oxidative stress. Changes in cell fluorescence were measured in a kinetic fashion using a confocal fluorescence microscope. The digital images were analyzed with dedicated software to measure the amount of oxidative stress experienced by each monolayer. Results: Values are means ± SEM.*p < 0.05 vs baseline, 1 p < 0.05 vs PFOB. Conclusions: PFOB attenuates oxidative injury to cell monolayers. We speculate that due to its high spreading coefficient, PFOB may act as a physical barrier against free radicals originated in the aqueous phase. The lipophilic PFOB could potentially alter cell membrane fluidity and confer some protection even after its removal. Supported, in part, by the American Lung Association- NY Affiliate, and by Alliance Pharmaceutical Corporation.",
author = "Rotta, {Alexandre T.} and Bj{\"o}rn Gunnarsson and Hernan, {Lynn J.} and Fuhrman, {Bradley P.} and Steinhorn, {David M.}",
year = "1999",
language = "English (US)",
volume = "27",
journal = "Critical Care Medicine",
issn = "0090-3493",
publisher = "Lippincott Williams and Wilkins",
number = "1 SUPPL.",

}

TY - JOUR

T1 - Perflubron (PFOB) protects cell monolayers against direct in vitro oxidative injury

AU - Rotta, Alexandre T.

AU - Gunnarsson, Björn

AU - Hernan, Lynn J.

AU - Fuhrman, Bradley P.

AU - Steinhorn, David M.

PY - 1999

Y1 - 1999

N2 - Introduction: Oxidative stress is often implicated in the genesis of injury to living tissues, even though intact cells generally possess sophisticated antioxidant systems. PFOB attenuates oxidative lung injury in vivo, at least in part due to a decrease in free radical release by macrophages. We hypothesized that PFOB would also protect a biological system from direct injury by reactive oxygen species. Methods: Rat pulmonary artery endothelial cells (RPAECs) were grown to confluence on optical grade culture dishes using standard technique. Cells were labeled with a permeable intracellular probe containing dihydrochlorofluorescein-diacetate (DCFH-DA), activated by cytoplasmic esterases. The cells were exposed to PFOB or culture medium for 15 min, after which all PFOB was removed by a transfer pipette. The monolayers were washed with medium prior to introduction of a buffer containing H2O2 to induce oxidative stress. Changes in cell fluorescence were measured in a kinetic fashion using a confocal fluorescence microscope. The digital images were analyzed with dedicated software to measure the amount of oxidative stress experienced by each monolayer. Results: Values are means ± SEM.*p < 0.05 vs baseline, 1 p < 0.05 vs PFOB. Conclusions: PFOB attenuates oxidative injury to cell monolayers. We speculate that due to its high spreading coefficient, PFOB may act as a physical barrier against free radicals originated in the aqueous phase. The lipophilic PFOB could potentially alter cell membrane fluidity and confer some protection even after its removal. Supported, in part, by the American Lung Association- NY Affiliate, and by Alliance Pharmaceutical Corporation.

AB - Introduction: Oxidative stress is often implicated in the genesis of injury to living tissues, even though intact cells generally possess sophisticated antioxidant systems. PFOB attenuates oxidative lung injury in vivo, at least in part due to a decrease in free radical release by macrophages. We hypothesized that PFOB would also protect a biological system from direct injury by reactive oxygen species. Methods: Rat pulmonary artery endothelial cells (RPAECs) were grown to confluence on optical grade culture dishes using standard technique. Cells were labeled with a permeable intracellular probe containing dihydrochlorofluorescein-diacetate (DCFH-DA), activated by cytoplasmic esterases. The cells were exposed to PFOB or culture medium for 15 min, after which all PFOB was removed by a transfer pipette. The monolayers were washed with medium prior to introduction of a buffer containing H2O2 to induce oxidative stress. Changes in cell fluorescence were measured in a kinetic fashion using a confocal fluorescence microscope. The digital images were analyzed with dedicated software to measure the amount of oxidative stress experienced by each monolayer. Results: Values are means ± SEM.*p < 0.05 vs baseline, 1 p < 0.05 vs PFOB. Conclusions: PFOB attenuates oxidative injury to cell monolayers. We speculate that due to its high spreading coefficient, PFOB may act as a physical barrier against free radicals originated in the aqueous phase. The lipophilic PFOB could potentially alter cell membrane fluidity and confer some protection even after its removal. Supported, in part, by the American Lung Association- NY Affiliate, and by Alliance Pharmaceutical Corporation.

UR - http://www.scopus.com/inward/record.url?scp=25044481081&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=25044481081&partnerID=8YFLogxK

M3 - Article

AN - SCOPUS:25044481081

VL - 27

JO - Critical Care Medicine

JF - Critical Care Medicine

SN - 0090-3493

IS - 1 SUPPL.

ER -