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Open Access

The effect of perturbations of the glycocalyx on microvascular perfusion in the obese trauma population: an in vitro study

Lawrence N Diebel, Alexander Liviu Marinica, David Edelman, David Liberati
DOI: 10.1136/tsaco-2021-000711 Published 26 April 2021
Lawrence N Diebel
Surgery, Wayne State University School of Medicine, Detroit, Michigan, USA
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Alexander Liviu Marinica
Surgery, Wayne State University School of Medicine, Detroit, Michigan, USA
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David Edelman
Surgery, Wayne State University School of Medicine, Detroit, Michigan, USA
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David Liberati
Surgery, Wayne State University School of Medicine, Detroit, Michigan, USA
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    Figure 1

    Experimental overview. epi, epinephrine; HUVEC, human umbilical vein endothelial cell; RBC, red blood cell.

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    Figure 2

    (A) Red blood cell (RBC) glycocalyx staining and fluorescence intensity measurement. RBC glycocalyx from patients with and without obesity was stained with fluorescein isothiocyanate-conjugated wheat germ agglutinin antibody after exposure to hypoxia/reoxygenation and epinephrine. Pictures were taken with a Leica TCS SP5 inverted fluorescent microscope and fluorescent intensity measurements were taken. (B) Shedding of RBC glycocalyx components. ELISA kits were used to quantitate syndecan-1 and hyaluronic acid levels present in RBC from patients with and without obesity exposed to hypoxia (1% O2) and epinephrine perfusion for 90 min.

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    Figure 3

    (A) Glycocalyx shedding in human umbilical vein endothelial cell (HUVEC) monolayers. ELISA kits were used to quantitate syndecan-1 and hyaluronic acid (HLA) levels present in the perfusate collected from HUVEC exposed to hypoxia (Hyp) (1% O2) and epinephrine (epi) perfusion for 90 min. (B) HUVEC glycocalyx thickness measurement. Three-dimensional XYZ image stacks were acquired and processed and analyzed using Volocity cellular imaging and analysis software to assess glycocalyx thickness.

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    Figure 4

    Red blood cell (RBC) adhesion to human umbilical vein endothelial cell (HUVEC) from subjects with and without obesity. RBCs from patients without obesity and RBCs from patients with obesity were added to the perfusate and allowed to flow through the microfluidic device. Adherent cells were photographed and counted using the Zeiss Observer Spinning Disk Confocal Microscope (Microscopy, Imaging and Cytometry core facility at Wayne State University, Detroit, Michigan, USA). *P<0.05 vs non-obese RBC, #p<0.05 vs same group no hypoxia (Hyp)+epinephrine (epi)).

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The effect of perturbations of the glycocalyx on microvascular perfusion in the obese trauma population: an in vitro study
Lawrence N Diebel, Alexander Liviu Marinica, David Edelman, David Liberati
Trauma Surg Acute Care Open Apr 2021, 6 (1) e000711; DOI: 10.1136/tsaco-2021-000711

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The effect of perturbations of the glycocalyx on microvascular perfusion in the obese trauma population: an in vitro study
Lawrence N Diebel, Alexander Liviu Marinica, David Edelman, David Liberati
Trauma Surg Acute Care Open Apr 2021, 6 (1) e000711; DOI: 10.1136/tsaco-2021-000711
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The effect of perturbations of the glycocalyx on microvascular perfusion in the obese trauma population: an in vitro study
Lawrence N Diebel, Alexander Liviu Marinica, David Edelman, David Liberati
Trauma Surgery & Acute Care Open Apr 2021, 6 (1) e000711; DOI: 10.1136/tsaco-2021-000711
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