Kinetics of plasma membrane electron transport in a pulmonary endothelial cell-column

L. E. Olson; M. P. Merker; R. D. Bongard; B. M. Brantmeier; S. H. Audi; J. H. Linehan; C. A. Dawson

doi:10.1114/1.83

Kinetics of plasma membrane electron transport in a pulmonary endothelial cell-column

L. E. Olson^*, M. P. Merker, R. D. Bongard, B. M. Brantmeier, S. H. Audi, J. H. Linehan, C. A. Dawson

^*Corresponding author for this work

Biomedical Engineering

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

5 Scopus citations

Abstract

Thiazine dyes such as toluidine blue O (TBO) are reduced at the luminal endothelial surface. The purpose of this study was to determine the rate of this reaction in endothelial cells in culture. A multiple indicator dilution method was used to measure the reaction kinetics during transient passage of a TBO-containing bolus through a chromatographic column filled with bovine pulmonary arterial endothelial cells grown on microcarrier beads (cell-column). A bolus containing TBO and an inert extracellular reference indicator (FITC-Dextran) was injected upstream of the cell-column, and the indicator concentrations were measured downstream using on-line photodetection. The effects of column flow rate, PO₂, and TBO concentration were studied. The fraction of TBO reduced upon passage through the cell-column decreased with increasing flow indicating that the reaction rate rather than TBO delivery controlled TBO reduction. The fraction of TBO reduced did not change with PO₂ or dose in the ranges studied. TBO reduction was about 10 times that for steady state TBO sequestration by these cells which, along with the lack of a PO₂ effect indicates that the rapid rate of reduction is not the rate-limiting step in steady state sequestration.

Original language	English (US)
Pages (from-to)	117-127
Number of pages	11
Journal	Annals of Biomedical Engineering
Volume	26
Issue number	1
DOIs	https://doi.org/10.1114/1.83
State	Published - 1998

Funding

This work was supported by NHBLI Grant Nos. HL24349 and HL52108, The Department of Veterans Affairs, The Whitaker Foundation, and The Howard Hughes Medical Institute. We also gratefully acknowledge the help of Eva T. Dembowski and Aimee H. Weit-zel for their experimental assistance.

Keywords

Cell-column chromatography
Endothelium
Oxidation-reduction
Toluidine blue O

ASJC Scopus subject areas

Biomedical Engineering

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10.1114/1.83

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@article{1890ffa8a69044cb852c791a5ea0804d,

title = "Kinetics of plasma membrane electron transport in a pulmonary endothelial cell-column",

abstract = "Thiazine dyes such as toluidine blue O (TBO) are reduced at the luminal endothelial surface. The purpose of this study was to determine the rate of this reaction in endothelial cells in culture. A multiple indicator dilution method was used to measure the reaction kinetics during transient passage of a TBO-containing bolus through a chromatographic column filled with bovine pulmonary arterial endothelial cells grown on microcarrier beads (cell-column). A bolus containing TBO and an inert extracellular reference indicator (FITC-Dextran) was injected upstream of the cell-column, and the indicator concentrations were measured downstream using on-line photodetection. The effects of column flow rate, PO2, and TBO concentration were studied. The fraction of TBO reduced upon passage through the cell-column decreased with increasing flow indicating that the reaction rate rather than TBO delivery controlled TBO reduction. The fraction of TBO reduced did not change with PO2 or dose in the ranges studied. TBO reduction was about 10 times that for steady state TBO sequestration by these cells which, along with the lack of a PO2 effect indicates that the rapid rate of reduction is not the rate-limiting step in steady state sequestration.",

keywords = "Cell-column chromatography, Endothelium, Oxidation-reduction, Toluidine blue O",

author = "Olson, {L. E.} and Merker, {M. P.} and Bongard, {R. D.} and Brantmeier, {B. M.} and Audi, {S. H.} and Linehan, {J. H.} and Dawson, {C. A.}",

note = "Funding Information: This work was supported by NHBLI Grant Nos. HL24349 and HL52108, The Department of Veterans Affairs, The Whitaker Foundation, and The Howard Hughes Medical Institute. We also gratefully acknowledge the help of Eva T. Dembowski and Aimee H. Weit-zel for their experimental assistance.",

year = "1998",

doi = "10.1114/1.83",

language = "English (US)",

volume = "26",

pages = "117--127",

journal = "Annals of Biomedical Engineering",

issn = "0090-6964",

publisher = "Springer Netherlands",

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T1 - Kinetics of plasma membrane electron transport in a pulmonary endothelial cell-column

AU - Olson, L. E.

AU - Merker, M. P.

AU - Bongard, R. D.

AU - Brantmeier, B. M.

AU - Audi, S. H.

AU - Linehan, J. H.

AU - Dawson, C. A.

N1 - Funding Information: This work was supported by NHBLI Grant Nos. HL24349 and HL52108, The Department of Veterans Affairs, The Whitaker Foundation, and The Howard Hughes Medical Institute. We also gratefully acknowledge the help of Eva T. Dembowski and Aimee H. Weit-zel for their experimental assistance.

PY - 1998

Y1 - 1998

N2 - Thiazine dyes such as toluidine blue O (TBO) are reduced at the luminal endothelial surface. The purpose of this study was to determine the rate of this reaction in endothelial cells in culture. A multiple indicator dilution method was used to measure the reaction kinetics during transient passage of a TBO-containing bolus through a chromatographic column filled with bovine pulmonary arterial endothelial cells grown on microcarrier beads (cell-column). A bolus containing TBO and an inert extracellular reference indicator (FITC-Dextran) was injected upstream of the cell-column, and the indicator concentrations were measured downstream using on-line photodetection. The effects of column flow rate, PO2, and TBO concentration were studied. The fraction of TBO reduced upon passage through the cell-column decreased with increasing flow indicating that the reaction rate rather than TBO delivery controlled TBO reduction. The fraction of TBO reduced did not change with PO2 or dose in the ranges studied. TBO reduction was about 10 times that for steady state TBO sequestration by these cells which, along with the lack of a PO2 effect indicates that the rapid rate of reduction is not the rate-limiting step in steady state sequestration.

AB - Thiazine dyes such as toluidine blue O (TBO) are reduced at the luminal endothelial surface. The purpose of this study was to determine the rate of this reaction in endothelial cells in culture. A multiple indicator dilution method was used to measure the reaction kinetics during transient passage of a TBO-containing bolus through a chromatographic column filled with bovine pulmonary arterial endothelial cells grown on microcarrier beads (cell-column). A bolus containing TBO and an inert extracellular reference indicator (FITC-Dextran) was injected upstream of the cell-column, and the indicator concentrations were measured downstream using on-line photodetection. The effects of column flow rate, PO2, and TBO concentration were studied. The fraction of TBO reduced upon passage through the cell-column decreased with increasing flow indicating that the reaction rate rather than TBO delivery controlled TBO reduction. The fraction of TBO reduced did not change with PO2 or dose in the ranges studied. TBO reduction was about 10 times that for steady state TBO sequestration by these cells which, along with the lack of a PO2 effect indicates that the rapid rate of reduction is not the rate-limiting step in steady state sequestration.

KW - Cell-column chromatography

KW - Endothelium

KW - Oxidation-reduction

KW - Toluidine blue O

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Kinetics of plasma membrane electron transport in a pulmonary endothelial cell-column

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