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October 15, 2006
2:00 PM - 4:00 PM
Room Hall E, Area L
The Effect of Cyclic Uniaxial Stretch on E-Selectin Expression in HUVEC Cells
Klaudiusz Suchodolski, M.D., Claudia T. Mierke, Ph.D., Monika Hedwig-Geissing, Ph.D., Jürgen Schüttler, M.D., Wolfgang H. Goldmann, Ph.D.
Department of Anesthesiology, Friedrich-Alexander-University Erlangen-Nuremberg, Erlangen, Germany
Background and Aim of Study : Ventilator-induced lung injury (VILI) has been shown to result from the mechanical stress of unstable lung units including endothelial cells. However, the biochemical and molecular pathways associated with mechanical stress-induced vascular injury remained lucid. E-selectin expressed on endothelium, mediates rolling of leukocytes and enhances the accumulation of leukocyte at sites of inflammation, leading to increased vascular permeability and tissue damage. Involvement of E-selectins in VILI has never directly been demonstrated. The aim of this study was to investigate whether mechanical stress (strain) applied to endothelial cells induces E-selectin expression.

Materials and Methods: Endothelial cells from a human umbilical cord vein (HUVEC) were cultured on an elastic silicone membrane coated with 5 µg/ml collagen. Using a stretch apparatus STREX ST-140-04 (B-Bridge, Sunnyvale, CA, USA) cells were subjected for 6 h or 24 h to uniaxial sinusoidal stretch (130% peak to peak at 11 Hz) at 37°C, 5% CO2 [Stretch (+)/TNF-α (-)]. For negative control, cells were identically cultured and treated except for the absence of uniaxial sinusoidal stretch [Stretch (-)/TNF-α (-)]. As positive control for E-selectin expression, cells were stimulated with 10 ng/ml TNF-α for 6 h and 24 h [Stretch (-)/TNF-α (+)]. E-selectin expression on HUVEC cells was measured by immunofluorescent flow cytometric analysis using a FACSCalibur cytometer (Becton Dickinson, Heidelberg, Germany). Morphological changes in cell structure and orientation were monitored microscopically in a time dependent manner.

Results: TNF-α treatment significantly upregulated E-selectin expression (Fig. 1). By contrast uniaxial stretch used in current experimental setting failed to augment E-selectin expression (Fig. 1). There were no significant morphological changes in cell structure/orientation after 6 h and 24 h of sinusoidal stretch.[figure1]

Anesthesiology 2006; 105: A731
Figure 1