The stressed vascular barrier and coagulation - The impact of key glycocalyx components on in vitro clot formation

应激血管屏障和凝血 - 关键糖萼成分对体外凝血形成的影响

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作者:Judith-Irina Buchheim, Max C Enzinger, Alexander Choukèr, Mathias Bruegel, Lesca Holdt, Markus Rehm

Conclusions

All EGX components interfere with clot formation and strength. HS mimics heparin effects in ROTEM® that confirm detectable alterations of standard coagulation tests.

Material and methods

The EGX components syndecan-1 (S1), hyaluronic acid (HA) and heparan sulfate (HS) were added to blood samples of 10 healthy male volunteers separately in three distinct concentrations to mimic three severity levels of in vitro EGX shedding. We analyzed spiked blood samples for leukocyte derived reactive oxygen species (ROS) release as a measure for innate immune activation and evaluated the impact on coagulation using clinical standard coagulation tests (SCTs) as well as rotational thrombelastometry (ROTEM®).

Methods

The EGX components syndecan-1 (S1), hyaluronic acid (HA) and heparan sulfate (HS) were added to blood samples of 10 healthy male volunteers separately in three distinct concentrations to mimic three severity levels of in vitro EGX shedding. We analyzed spiked blood samples for leukocyte derived reactive oxygen species (ROS) release as a measure for innate immune activation and evaluated the impact on coagulation using clinical standard coagulation tests (SCTs) as well as rotational thrombelastometry (ROTEM®).

Results

Whereas ROS formation by polymorphonuclear leukocytes (PMN) was unaltered by all three substances, high concentrations of HS showed prolonged aPTT and TT compared to controls and S1 or HA. Changes in ROTEM® were discrete and mostly within normal range of values but analyses showed a significant reduction of clot firmness and formation by all EGX components compared to controls. Furthermore, alterations by HA and HS were dose dependent. Only HS showed a heparin like effect supporting the findings of SCTs. Conclusions: All EGX components interfere with clot formation and strength. HS mimics heparin effects in ROTEM® that confirm detectable alterations of standard coagulation tests.

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