BackgroundThe ischemia-reperfusion injury (IRI) is unavoidable in vascular surgery. Damage to the microcirculation and endothelial glycocalyx might set up a shock with loss of circulatory coherence and organ failure. Sulodexide may help to protect endothelial glycocalyx and alleviate the ischemia-reperfusion injury.MethodsTwenty female piglets underwent surgery with a 30-min-long suprarenal aortic clamp, followed by two hours of reperfusion. Ten piglets received sulodexide before the clamp, and 10 received normal saline. Blood and urine samples were taken at baseline and in 20-min intervals until the 120th minute to analyze the serum syndecan-1, E-selectin, and thrombomodulin. Albumin and glycosaminoglycans were examined in the urine. The kidney biopsies before and after the protocol were examined by light microscopy with hematoxylin-eosin staining. The sublingual microcirculation was recorded by side-stream dark field imaging at the time as blood and urine.ResultsBased on the 2-way ANOVA testing, there was no statistically significant difference in the parameters of sublingual microcirculation. Serum markers of endothelial cell activation and damage (E-selectin and thrombomodulin) did not show any statistically significant difference either. Syndecan-1, a marker of glycocalyx damage, showed statistically significantly higher values based on the 2-way ANOVA testing (p < 0.0001) with the highest difference in the 80th minute: 7.8 (3.9-44) ng/mL in the control group and 1.8 (0.67-2.8) ng/mL in the sulodexide group. In the urine, the albuminuria was higher in the control group, although not statistically significant. Glycosaminoglycans were statistically significantly higher in the sulodexide group based on the mixed-effect analysis due to the intervention itself. Histological analysis of the renal biopsies showed necrosis in both groups after reperfusion.ConclusionAdministering sulodexide significantly reduced the level of endothelial markers of IRI. The study results support further research into using preemptive administration of sulodexide to modulate IRI in clinical medicine.

• MeSH
• E-Selectin blood   MeSH
• Glycocalyx MeSH
• Glycosaminoglycans * pharmacology   therapeutic use   MeSH
• Kidney pathology   blood supply   MeSH
• Microcirculation drug effects   MeSH
• Disease Models, Animal MeSH
• Swine MeSH
• Reperfusion Injury * prevention & control   MeSH
• Syndecan-1 blood   MeSH
• Thrombomodulin blood   MeSH
• Animals MeSH
• Check Tag
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

BackgroundSulodexide is a glycosaminoglycan-based drug prescribed to patients with angiopathy. We performed a pilot study to investigate whether sulodexide positively modulates the endothelial glycocalyx (EG) layer and the microcirculation in a porcine model of EG enzymatic damage. The EG is a sugar-based endothelial lining that is involved in the physiology of the capillary wall and the pathogenesis of many diseases.MethodsEG damage was induced in eight piglets by hyaluronidase III and heparanase I given intravenously. Four animals received sulodexide 600 IU intravenously before the enzymes and four animals after the enzymes were administered. Four animals constituted a control group. Sublingual microcirculation by side-stream dark field imaging and plasmatic concentration of syndecan-1 by ELISA were measured at baseline, 20 min after intervention, and at the 40th, and 60th minute onwards. The statistics were performed with a one-way ANOVA test with Turkey's correction for multiple comparisons testing. Timepoint comparison was performed by Student t-test or Mann-Whitney test.ResultsAt baseline, there were no statistically significant differences between the animal groups. After the intervention, the levels of syndecan-1 were significantly lower in the control group. While there were no differences between the two intervention groups. The sublingual microcirculation analysis showed that the DeBacker score was significantly higher in the control group. At 60 min, there was also a statistically significant difference in DeBacker score between the groups (8.1 ± 1.6 mm-1 in the group with enzymes given first and 11 ± 0.92 mm-1 in the group with sulodexide given first, p = 0.03). The analysis of the proportion of perused vessels did not show any statistically significant differences.ConclusionThe results of the study demonstrated a working model of EG damage but no specific action of sulodexide on EG modulation. In the sublingual microcirculation analysis, the sulodexide reduced the fall in absolute tissue perfusion in 60 min.

• MeSH
• Endothelium, Vascular * drug effects   MeSH
• Glycocalyx * drug effects   metabolism   MeSH
• Glycosaminoglycans * pharmacology   MeSH
• Hyaluronoglucosaminidase MeSH
• Microcirculation drug effects   MeSH
• Disease Models, Animal MeSH
• Pilot Projects MeSH
• Swine MeSH
• Syndecan-1 blood   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH

BACKGROUND: Excess fluid in the interstitium can adversely affect the microcirculation. We studied how gradual dilution of the blood plasma by crystalloid fluid influences microcirculatory variables and capillary filtration in 20 patients undergoing surgery. METHODS: Video recordings of the sublingual mucosal were made on four occasions during the surgery and compared with quasi-measurements of the capillary filtration rate using retrospective volume kinetic data collected over 5-10-minute periods during 262 infusion experiments with crystalloid fluid. RESULTS: The number of crossings (vessel density) increased up to plasma dilution of 15-20 % whereafter it decreased. The proportion of the vessels that were perfused (PPV) decreased and reached a nadir of -15 % at a dilution of 20-30 %. Changes in the number of crossings and the PPV correlated (r = 0.62, P < 0.001) but the curve was displaced so that crossings showed no change when PPV had decreased by approximately 10 %. However, the PPV of vessels with a thickness of ≤25 μm increased or remained constant in the dilution range of up to 20 %. The volume kinetic analysis showed that the capillary filtration was greater than expected from proportionality with the volume expansion up to a plasma dilution of 15 %, the greatest difference (+89 %) being for plasma dilution up to 5 %. CONCLUSION: Plasma dilution of up to 15 % increased the vessel density, and the capillary filtration increased by more than suggested by the volume expansion. Dilution >15 % had a negative influence on these variables.

• MeSH
• Video Recording MeSH
• Time Factors MeSH
• Adult MeSH
• Hemodilution * MeSH
• Isotonic Solutions administration & dosage   MeSH
• Capillary Permeability MeSH
• Capillaries physiopathology   MeSH
• Kinetics MeSH
• Crystalloid Solutions * administration & dosage   MeSH
• Middle Aged MeSH
• Humans MeSH
• Microcirculation * MeSH
• Regional Blood Flow MeSH
• Retrospective Studies MeSH
• Blood Flow Velocity MeSH
• Aged MeSH
• Mouth Mucosa blood supply   MeSH
• Mouth Floor blood supply   MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Introduction: Diabetic ketoacidosis (DKA) is associated with volume depletion and hemodynamic alterations. Changes in systemic microcirculation during DKA have not been described so far. Methods: In this case report, we describe the evolution of sublingual microcirculatory changes, monitored using sidestream dark field (SDF) imaging during the treatment of severe diabetic ketoacidosis in a 13-year-old girl. The patient presented a pH of 6.84, a glycemia level of 27.2 mmol/L, a ketonemia level of 5.6 mmol/L, a base excess of -29.4 mmol/L, hypernatremia, hyperosmolality due to acute gastritis, and a malfunction of the glucose sensor. Sublingual microcirculation measurements using an SDF probe were initiated 60 min after the initiation of treatment, which was then repeated 2, 3, 4, 6, 12, and 24 h after treatment initiation, as well as on the day of discharge. Results: Substantial alterations of microvascular perfusion parameters, both total and small vessel densities, perfused vessel densities, and the DeBacker score, were observed during the first 6 to 12 h of treatment. The degree of microcirculatory alteration was strongly negatively correlated with calculated osmolality, sodium levels, ketone and lactate levels, and blood pressure values. Conclusions: DKA is, in its complexity, associated with a serious microcirculatory alteration. SDF imaging provides insight into the severity of the patient's microcirculatory alteration and its evolution during treatment.

• Publication type
• Case Reports MeSH

BACKGROUND: Hydrogen is a potent antioxidant agent that can easily be administered by inhalation. The aim of the study was to evaluate whether hydrogen protects the endothelial glycocalyx layer after successful cardiopulmonary resuscitation (CPR). METHODS: Fourteen anesthetized pigs underwent CPR after induced ventricular fibrillation. During CPR and return of spontaneous circulation, 2% hydrogen gas was administered to seven pigs (hydrogen group) and seven constituted a control group. Biochemistry and sublingual microcirculation were assessed at baseline, during CPR, at the 15th, 30th, 60th, 120th minute. RESULTS: All seven subjects from the hydrogen group and six subjects in the control group were successfully resuscitated after 6-10 minutes. At baseline, there were no statistically significant differences in examined variables. After the CPR, blood pH, base excess, and lactate showed significantly smaller deterioration in the hydrogen group than in the control group. By contrast, plasma syndecan-1 and the measured variables obtained via sublingual microcirculation did not change after the CPR; and were virtually identical between the two groups. CONCLUSION: In pigs, hydrogen gas inhalation during CPR and post-resuscitation care was associated with less pronounced metabolic acidosis compared to controls. However, we could not find evidence of injury to the endothelium or glycocalyx in any studied groups.

• MeSH
• Endothelium MeSH
• Glycocalyx MeSH
• Cardiopulmonary Resuscitation * MeSH
• Humans MeSH
• Disease Models, Animal MeSH
• Swine MeSH
• Reperfusion Injury * MeSH
• Heart Arrest * therapy   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

BACKGROUND: The sidestream dark-field imaging method is used to study microcirculation. Normal values of sublingual microcirculation parameters in healthy children of different age and gender categories are unknown. OBJECTIVE: The study's main goal was to determine normal values of selected parameters of sublingual microcirculation in healthy children of different age and gender categories. METHODS: 40 healthy children were measured, ten aged 3-5.9 years, ten aged 6-10.9 years, ten aged 11-14.9 years, and ten aged 15-18.9 years. After recording the basic anthropometric parameters and vital functions, each volunteer had their microcirculation measured using an SDF probe placed sublingually. Three video clips were recorded and processed offline, and the three best and most stable parts of each were analyzed. RESULTS: Total vascular density, small vessel density, proportion of perfused small vessels, perfused vessel density, perfused small vessel density, and DeBacker's score were significantly higher in females than in males. There were no differences between age groups in microcirculation parameters except MFI. CONCLUSIONS: Age does not influence normal values of microcirculatory parameters. Female gender was associated with higher vessel density, perfused vessel density, and DeBacker's score. A suggestion of the normal range of microcirculatory parameters in healthy children is provided.

• MeSH
• Child MeSH
• Volunteers * MeSH
• Humans MeSH
• Microcirculation MeSH
• Mouth Floor * blood supply   MeSH
• Check Tag
• Child MeSH
• Humans MeSH
• Male MeSH
• Female MeSH
• Publication type
• Journal Article MeSH

BACKGROUND: Coronavirus disease (COVID-19) associated endotheliopathy and microvascular dysfunction are of concern. OBJECTIVE: The objective of the present single-center observational pilot study was to compare endothelial glycocalyx (EG) damage and endotheliopathy in patients with severe COVID-19 (COVID-19 group) with patients with bacterial pneumonia with septic shock (non-COVID group). METHODS: Biomarkers of EG damage (syndecan-1), endothelial cells (EC) damage (thrombomodulin), and activation (P-selectin) were measured in blood on three consecutive days from admission to the intensive care unit (ICU). The sublingual microcirculation was studied by Side-stream Dark Field (SDF) imaging with automatic assessment. RESULTS: We enrolled 13 patients in the non-COVID group (mean age 70 years, 6 women), and 15 in the COVID-19 group (64 years old, 3 women). The plasma concentrations of syndecan-1 were significantly higher in the COVID-19 group during all three days. Differences regarding other biomarkers were not statistically significant. The assessment of the sublingual microcirculation showed improvement on Day 2 in the COVID-19 group. Plasma levels of C-reactive protein (CRP) were significantly higher on the first two days in the COVID-19 group. Plasma syndecan-1 and CRP were higher in patients suffering from severe COVID-19 pneumonia compared to bacterial pneumonia patients. CONCLUSIONS: These findings support the role of EG injury in the microvascular dysfunction in COVID-19 patients who require ICU.

• MeSH
• Biomarkers MeSH
• COVID-19 * pathology   MeSH
• Endothelial Cells * pathology   MeSH
• Glycocalyx * metabolism   MeSH
• Middle Aged MeSH
• Humans MeSH
• Pilot Projects MeSH
• Prospective Studies MeSH
• Aged MeSH
• Syndecan-1 metabolism   MeSH
• Respiration, Artificial MeSH
• Check Tag
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Observational Study MeSH

BACKGROUND: The endothelial glycocalyx (EG) plays a vital role in the physiology and pathophysiology of human microcirculation. Having relevant EG damage model would be important tool for testing new interventions aiming at EG protection and recovery. We describe the first in vivo EG damage model in pig. OBJECTIVE: To investigate the course of animal EG damage induced by specific enzymes. MATERIAL AND METHODS: Four anesthetized piglets received enzymes: 1g hyaluronidase and 25 IU heparanase I intravenously. Blood and urine samples were collected at baseline and 20/40/60/80/100/120 min for detecting markers of endothelial and EG function. Sublingual microcirculation and EG thickness were assessed by Side-stream Dark Field (SDF) imaging and Perfused Boundary Region (PBR) respectively. EG of the mesentery artery was visualized in fluorescent microscopy. RESULTS: Biochemical marker of EG damage syndecan-1 showed temporary increase with return to baseline and was reflected by PBR values. Albumin levels suggested brief period of capillary leakage (decrease in the serum, increase in the urine) with a trend to normalization. Urine glycosaminoglycans peaked at 120 minutes. Microcirculatory perfusion parameter showed significant alteration. Diffusion parameters were altered with no statistical significance. CONCLUSION: EG damage induced by specific enzymes was reflected by temporary changes of biochemical makers together with alteration of microcirculation and changes in fluorescent microscopy of EG layer. Our results support to further validate presented model of EG damage on a larger number of animals.

• MeSH
• Glycocalyx * MeSH
• Capillaries MeSH
• Microcirculation MeSH
• Pilot Projects MeSH
• Swine MeSH
• Digestion MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH

Cíl studie: Endoteliální glykokalyx (EG) hraje ústřední roli v udržení vaskulární integrity a správné funkce mikrocirkulace. Poškození EG u pacientů po úspěšné kardiopulmonální resuscitaci bylo popsáno měřením hladin rozpadových produktů EG (syndekan-1, heparan sulfát). Poměrně novou metodou k hodnocení stavu EG je parametr perfundované hraniční zóny (PBR), který udává v µm boční rozptyl červených krvinek směrem k EG. Cílem této studie bylo zhodnotit změny PBR na zvířecím modelu srdeční zástavy u prasete domácího a zhodnotit použitelnost PBR jako indikátoru poškození EG. Typ studie: Experimentální na zvířecím modelu srdeční zástavy u prasete domácího. Materiál a metody: Srdeční zástava byla indukována nitrosrdeční elektrodou navozením fibrilace komor, která byla léčena defibrilací dle doporučení ERC 2015 po 15 minutách. Sublingvální mikrocirkulace byla vyšetřena ručním mikroskopem fungujícím na principu ortogonálně polarizované spektroskopie. Pořízené nahrávky byly hodnoceny automaticky programem, který poskytl hodnotu PBR. Hodnocení bylo ve třech časových bodech: základní měření (PBR_B), po návratu spontánní cirkulace (PBR_ROSC) a 20 minut poté (PBR_ROSC_20). Výsledky: Hodnoty PBR byly získány od 11 prasat. Hodnoty PBR po ROSC nebyly statisticky signifikantně zvýšeny (p = 0,47). PBR_B bylo 2,129 (± 0,21) µm, PBR_ROSC bylo 2,206 (± 0,27) µm and PBR_ROSC_20 bylo 2,18 (± 0,19) µm. Výsledky jsou ve tvaru průměr (směrodatná odchylka). Závěr: Naše data udávají nevýznamné zvýšení parametru PBR v sublingvální oblasti po srdeční zástavě u prasete. Parametr PBR vyžaduje další testování, než by mohl být zaveden jako neinvazivní parametr udávající míru poškození EG na modelu srdeční zástavy u prasete.

Objective: Endothelial glycocalyx (EG) plays key role in maintaining vascular integrity. Perturbation of the EG in patients after cardiac arrest has been described by measuring syndecan-1 and heparan sulphate levels. Non-invasive method of evaluating EG thickness by using Perfused Boundary Region (PBR) has been introduced recently. PBR represents the amount of lateral deviation of red blood cells towards the EG in µm. The aim of the study was to evaluate changes in PBR on porcine model of cardiac arrest in order to assess the usefulness of PBR as an indicator of glycocalyx damage. Design: Experimental study on a porcine model of cardiac arrest. Material and Methods: Cardiac arrest was induced by intracardiac electrode triggering ventricular fibrillation and treated by defibrillation after 15 minutes in accordance with 2015 ERC guidelines. Sublingual microcirculation was measured by hand-held microscope working on the principle of orthogonal spectral imaging and PBR was computed automatically with specialized software in three timepoints. At baseline (PBR_B), after return of spontaneous circulation (PBR_ROSC) and 20 minutes after ROSC (PBR_ROSC_20). Results: PBR data was obtained from 11 pigs. There was insignificant increase of PBR after ROSC (p = 0.47). The PBR_B was 2.129 (± 0.21), PBR_ROSC was 2.206 (± 0.27) and PBR_ROSC_20 was 2.18 (± 0.19), data are presented as mean and standard deviation. Conclusions: Our data demonstrate insignificant increase of PBR after cardiac arrest in pig. The value of PBR as an indicator of EG shedding requires further study before introducing this parameter as a routine non-invasive tool in pig model of cardiac arrest.

• Keywords
• endoteliální glykokalyx,
• MeSH
• Endothelium, Vascular MeSH
• Glycocalyx * MeSH
• Cardiopulmonary Resuscitation MeSH
• Disease Models, Animal MeSH
• Death, Sudden, Cardiac * MeSH
• Swine MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Research Support, Non-U.S. Gov't MeSH

BACKGROUND: Endothelial glycocalyx (EG) is a carbohydrate-rich gel-like mesh covering the apical surface of endothelial cells. It has been linked to the microvascular pathophysiology and tissue metabolism. However, little is known about its condition in young healthy adults. OBJECTIVE: We aimed to describe the condition of EG in young healthy adults by in vivo EG imaging and measurement of syndecan-1, a plasma marker of EG integrity in order to obtain reference values. METHODS: For in vivo EG studies we used Side-stream Dark Field imaging of the sublingual microcirculation. Recordings were analysed automatically by GlycoCheck software providing the Perfused Boundary Region (PBR) as a marker of EG thickness. Levels of syndecan-1 were analysed in plasma samples by ELISA. RESULTS: 21 volunteers were included in the study. Median of the PBR value was 1.82 μm (interquartile range 1.69-2.01, 95% CI 1.79-1.97). Median concentration of syndecan-1 was 0.3 ng/ml (interquartile range 0.23-0.39, 95% CI 0.27-0.49). CONCLUSION: This study provides a comparison for cohorts of patients with a particular disease where the EG is presumably damaged. Our findings do not entirely comply with already published data in healthy individuals.

• MeSH
• Biomarkers metabolism   MeSH
• Adult MeSH
• Endothelial Cells metabolism   MeSH
• Glycocalyx metabolism   MeSH
• Humans MeSH
• Young Adult MeSH
• Healthy Volunteers MeSH
• Check Tag
• Adult MeSH
• Humans MeSH
• Young Adult MeSH
• Male MeSH
• Female MeSH
• Publication type
• Journal Article MeSH