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In vivo Measurement of the Mouse Pulmonary Endothelial Surface Layer
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Changes in endothelial glycocalyx layer protective ability after inflammatory stimulus.

Luis F Delgadillo1, Elena B Lomakina1, Julia Kuebel1

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Damage to the endothelial glycocalyx layer (EGL) increases leukocyte adhesion and endothelial barrier dysfunction. Removing both hyaluronic acid (HA) and heparan sulfate (HS) components significantly impacts neutrophil adhesion and fluid leakage.

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Area of Science:

  • Endothelial biology
  • Immunology
  • Sepsis research

Background:

  • Leukocyte adhesion to endothelium is critical in sepsis initiation and progression.
  • The endothelial glycocalyx layer (EGL) plays a role in neutrophil adhesion and barrier function, but research is limited.

Purpose of the Study:

  • To investigate the hypothesis that inflammation-induced EGL damage increases leukocyte adhesion and endothelial barrier dysfunction.
  • To model EGL damage using human umbilical vein endothelial cells with enzymatic removal of EGL components.

Main Methods:

  • Enzymatic removal of hyaluronic acid (HA) and heparan sulfate (HS) from EGL.
  • Atomic force microscopy to measure EGL thickness.
  • Assessment of neutrophil adhesion.
  • Measurement of transendothelial electrical resistance (TEER) and dextran diffusion to evaluate barrier properties.

Main Results:

  • Removal of HA or HS individually reduced EGL thickness; combined removal caused the most significant decrease.
  • Neutrophil adhesion increased only when both HA and HS were removed.
  • EGL component removal decreased TEER, indicating increased potential for fluid leakage, but did not significantly affect dextran diffusion.

Conclusions:

  • EGL damage during inflammation significantly affects adhesion molecule accessibility, promoting leukocyte adhesion.
  • Disruption of the EGL may contribute to increased fluid transport into tissues, exacerbating endothelial barrier dysfunction in sepsis.