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Intermittent flow increases endotoxin-induced adhesion of human erythrocytes to vascular endothelial cells

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Abstract

Objective

To investigate the effects of different conditions of flow on endotoxin induced adhesion of human red blood cells (RBC) to human umbilical vein endothelial cells (HUVEC).

Design and setting

Prospective, randomized, controlled in vitro study in a university-affiliated cell biology laboratory.

Subjects

Human erythrocytes, human vascular endothelial cells.

Interventions

Superfusion of HUVEC monolayers with human erythrocytes incubated with either saline (CON) or endotoxin (ETX) with different flow pattern (basic flow rates of 0.65 or 1.3 ml/min; intermittent flow, IMF). The CON/0.6, CON/1.3, CON-IMF/1.3 (n=7/group) groups served as control, and in test groups ETX/0.6, ETX/1.3, ETX-IMF/0.6, and ETX-IMF/1.3 (n=7/group) both RBC and HUVECs were incubated with ETX and flow pattern and rates varied. In the IMF experiments flow rates of 0.65 and 1.3 ml/min were combined with stop-and-go flow pattern.

Measurements and results

At continuous flow of 0.65 ml/min erythrocyte adhesion was 61±5 cells/mm2 in CON and 172±25 cells/mm2 after ETX. When flow rate was increased to 1.3 ml/min, adhesion decreased to 27±4 cells/mm2 in CON and 93±18 cells/mm2 after ETX. IMF conditions had no effect on RBC adhesion of naive RBC but increased the number of adhesive erythrocytes after incubation with ETX both at 0.65 ml/min (287±33 cells/mm2) and at 1.3 ml/min (148±13 cells/mm2).

Conclusions

RBC adhesion to vascular endothelium is affected by rate and pattern of blood flow. Higher flow rates or shear forces reduce RBC adhesion while stop-and-go flow pattern favored adhesion of ETX-treated erythrocytes to HUVECs. These findings suggest that altered RBCs interact with altered flow patterns potentially contributing to the microcirculatory injury observed in sepsis.

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Acknowledgements

The authors thank Ron Noseworthy for his assistance in cell harvesting, Dr. Natale for his cooperation in tissue collection, and the nurses of the delivery ward of St. Joseph's Hospital for their generous help in collecting umbilical cords.

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Authors and Affiliations

  1. A.C. Burton Vascular Biology Laboratory, London Health Sciences Centre, University of Western Ontario, 800 Commissoners Road East, London, Ontario, N6A 4G5, Canada

    Otto Eichelbrönner, William J. Sibbald & Ian H. Chin-Yee

  2. Klinik für Anästhesiologie, Bayerische Julius-Maximilians-Universität, Josef-Schneider-Strasse 2, 97080, Würzburg, Germany

    Otto Eichelbrönner

Authors
  1. Otto Eichelbrönner
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  2. William J. Sibbald
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  3. Ian H. Chin-Yee
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Corresponding author

Correspondence to Ian H. Chin-Yee.

Additional information

This research was supported by the Departments of Hematology and Critical Care Medicine, London Health Sciences Centre, London, Ontario, Canada.

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Eichelbrönner, O., Sibbald, W.J. & Chin-Yee, I.H. Intermittent flow increases endotoxin-induced adhesion of human erythrocytes to vascular endothelial cells. Intensive Care Med 29, 709–714 (2003). https://doi.org/10.1007/s00134-003-1698-y

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  • DOI: https://doi.org/10.1007/s00134-003-1698-y

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