Endothelial Glycocalyx Degradation Protocols

The endothelial glycocalyx was degraded by the infusion of hyaluronidase from bovine testes (25 mg/kg/body weight; H3884, Sigma, St. Louis, MO, USA) and a blend of heparinase I and III from Flavobacterium heparinum (1 U/mouse; H3917, Sigma, St. Louis, MO, USA) for 30 min. Heparinase units are given as Sigma units. The enzymes were prepared in phosphate-buffered saline (PBS) and were infused through femoral vein cannulation. The control animals were infused with PBS alone for 30 min. We have successfully demonstrated in a previous study a significant degradation of the mouse retinal glycocalyx following hyaluronidase infusion at a dose of 12 mg/kg [17 (link)].

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Kaur G., Leskova W, & Harris N.R. (2022). The Endothelial Glycocalyx and Retinal Hemodynamics. Pathophysiology, 29(4), 663-677.

Publication 2022

H3884 H3917

Animals Body weight Bovine Cannulation Endothelial Enzymes Femoral vein Flavobacterium heparinum Glycocalyx Heparinase Hyaluronidase Mouse Phosphate Retinal Saline Testes

Corresponding Organization : Louisiana State University Health Sciences Center Shreveport

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Variable analysis

independent variables

Infusion of hyaluronidase from bovine testes (25 mg/kg/body weight; H3884, Sigma, St. Louis, MO, USA)
Infusion of a blend of heparinase I and III from Flavobacterium heparinum (1 U/mouse; H3917, Sigma, St. Louis, MO, USA)

dependent variables

Degradation of the endothelial glycocalyx

control variables

Infusion of phosphate-buffered saline (PBS) alone for 30 min

controls

Positive control: Infusion of hyaluronidase at a dose of 12 mg/kg, which demonstrated significant degradation of the mouse retinal glycocalyx in a previous study [17].
Negative control: Infusion of PBS alone for 30 min.

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