Endothelial Glycocalyx Thickness Research Articles

Long-term efficacy of liposomal nanocarriers of preassembled glycocalyx in restoring cerebral endothelial glycocalyx in sepsis

The endothelial glycocalyx (EG) undergoes early degradation in sepsis. Our recent work introduced a novel therapeutic approach involving liposomal nanocarriers of preassembled glycocalyx (LNPG) to restore EG in lipopolysaccharide (LPS)-induced sepsis model of mice. While short-term effects were promising, this study focuses on the long-term impact of LNPG on mouse cerebral microcirculation. Utilizing cranial window, we assessed the stability of vascular density (VD) and perfused boundary region (PBR), an index of EG thickness, over a five-day period in normal control mice. In septic groups (LPS, LPS + 1-dose LNPG, and LPS + 2-dose LNPG), the exposure of mice to LPS significantly reduced VD and increased PBR within 3 h. Without LNPG treatment, PBR returned to the normal control level by endogenous processes at 48 h, associated with the recovery of VD to the baseline level at 72 h. However, mice receiving LNPG treatment significantly reduced the increment of PBR at 3 h. The therapeutic effect of 1-dose LNPG persisted for 6 h while the 2-dose LNPG treatment further reduced PBR and significantly increased VD at 12 h compared to LPS group. This study provides valuable insights into the potential therapeutic benefits of LNPG in mitigating EG degradation in sepsis.

Assessment of the endothelial glycocalyx state in patients with axial spondyloarthritis associated with Crohn’s disease

BACKGROUND: Epithelial barrier dysfunction, including their permeability increment, is supposed to be a pathogenetic link in the development of various immunoinflammatory diseases, including Crohn’s disease and axial spondyloarthritits. Endothelial permeability regulation is provided by both intercellular contacts and endothelial glycocalyx. AIM: To study the microcirculatory state by measuring the endothelial glycocalyx thickness in patients with axial spondyloarthritits associated with Crohn’s disease. To identify an association between the endothelial glycocalyx thinning and the epithelial permeability severity in patients with Crohn’s disease, axial spondyloarthritits and their combination. MATERIALS AND METHODS: 12 patients with axial spondyloarthritits associated with Crohn’s disease (group A), 22 patients with idiopathic axial spondyloarthritits (group B), 24 patients with Crohn’s disease (group C) and 16 healthy individuals (group D) were examined. The endothelial glycocalyx thickness was assessed in the sublingual region using dark-field microscopy. Fecal calprotectin and fecal zonulin was studied. RESULTS: A statistically significant increase in the perfused boundary region high flow was found (p = 0.001), as well as the MicroVascular Health Score (p = 0.004) in group A compared with the control. In Group A, a positive correlation between perfused boundary region high flow and scores Ankylosing Spondylitis Disease Activity Score, Bath Ankylosing Spondylitis Disease Activity Index, Bath Ankylosing Spondylitis Metrology Index 10 (p 0.05) was found; perfused boundary region 5–9 mµ correlated positively with the fecal calprotectin (p = 0.007). Perfused boundary region high flow measurement provides 77.1% accuracy for correct classification of axial spondyloarthritits associated with Crohn’s disease and idiopathic axial spondyloarthritits (p = 0.010). In group C, a positive correlation between fecal zonulin and perfused boundary region 5–9 mµ, 10–19 mµ, 5–25 mµ (p 0.05) was also found. CONCLUSIONS: The findings suggest that the patients with axial spondyloarthritits associated with Crohn’s disease have endothelial glycocalyx impairment; the detection of these abnormalities may be useful in assessing the disease integral activity and in the differential diagnosis of axial spondyloarthritits associated with Crohn’s disease and idiopathic axial spondyloarthritits. Further studies of microcirculation will allow better understanding of spondyloarthritis and inflammatory bowel diseases pathogenesis and improve the management of patients with these diseases.

The association between prolonged capillary refill time and microcirculation changes in children with sepsis

BackgroundsIn children with sepsis, circulatory shock and multi-organ failure remain major contributors to mortality. Prolonged capillary refill time (PCRT) is a clinical tool associated with disease severity and tissue hypoperfusion. Microcirculation assessment with videomicroscopy represents a promising candidate for assessing and improving hemodynamic management strategies in children with sepsis. Particularly when there is loss of coherence between the macro and microcirculation (hemodynamic incoherence). We sought to evaluate the association between PCRT and microcirculation changes in sepsis.MethodsThis was a prospective cohort study in children hospitalized with sepsis. Microcirculation was measured using sublingual video microscopy (capillary density and flow and perfused boundary region [PBR]—a parameter inversely proportional to vascular endothelial glycocalyx thickness), phalangeal tissue perfusion, and endothelial activation and glycocalyx injury biomarkers. The primary outcome was the association between PCRT and microcirculation changes.ResultsA total of 132 children with sepsis were included, with a median age of two years (IQR 0.6–12.2). PCRT was associated with increased glycocalyx degradation (PBR 2.21 vs. 2.08 microns; aOR 2.65, 95% CI 1.09–6.34; p = 0.02) and fewer 4–6 micron capillaries recruited (p = 0.03), with no changes in the percentage of capillary blood volume (p = 0.13). Patients with hemodynamic incoherence had more PBR abnormalities (78.4% vs. 60.8%; aOR 2.58, 95% CI 1.06–6.29; p = 0.03) and the persistence of these abnormalities after six hours was associated with higher mortality (16.5% vs. 6.1%; p < 0.01). Children with an elevated arterio-venous CO2 difference (DCO2) had an abnormal PBR (aOR 1.13, 95% CI 1.01–1.26; p = 0.03) and a lower density of small capillaries (p < 0.05). Prolonged capillary refill time predicted an abnormal PBR (AUROC 0.81, 95% CI 0.64–0.98; p = 0.03) and relative percentage of blood in the capillaries (AUROC 0.82, 95% CI 0.58–1.00; p = 0.03) on admission. A normal CRT at 24 h predicted a shorter hospital stay (aOR 0.96, 95% CI 0.94–0.99; p < 0.05).ConclusionsWe found an association between PCRT and microcirculation changes in children with sepsis. These patients had fewer small capillaries recruited and more endothelial glycocalyx degradation. This leads to nonperfused capillaries, affecting oxygen delivery to the tissues. These disorders were associated with hemodynamic incoherence and worse clinical outcomes when the CRT continued to be abnormal 24 h after admission.

Sedation with propofol and isoflurane differs in terms of microcirculatory parameters: A randomized animal study using dorsal skinfold chamber mouse model

ObjectiveThis study aimed to explore the effects of sedative doses of propofol and isoflurane on microcirculation in septic mice compared to controls. Isoflurane, known for its potential as a sedation drug in bedside applications, lacks clarity regarding its impact on the microcirculation system. The hypothesis was that propofol would exert a more pronounced influence on the microvascular flow index, particularly amplified in septic conditions. Material and methodsRandomized study was conducted from December 2020 to October 2021 involved 60 BALB/c mice, with 52 mice analyzed. Dorsal skinfold chambers were implanted, followed by intraperitoneal injections of either sterile 0.9 % saline or lipopolysaccharide for the control and sepsis groups, respectively. Both groups received propofol or isoflurane treatment for 120 min. Microcirculatory parameters were obtained via incident dark-field microscopy videos, along with the mean blood pressure and heart rate at three time points: before sedation (T0), 30 min after sedation (T30), and 120 min after sedation (T120). Endothelial glycocalyx thickness and syndecan-1 concentration were also analyzed. ResultsIn healthy controls, both anesthetics reduced blood pressure. However, propofol maintained microvascular flow, differing significantly from isoflurane at T120 (propofol, 2.8 ± 0.3 vs. isoflurane, 1.6 ± 0.9; P < 0.001). In the sepsis group, a similar pattern occurred at T120 without statistical significance (propofol, 1.8 ± 1.1 vs. isoflurane, 1.2 ± 0.7; P = 0.023). Syndecan-1 levels did not differ between agents, but glycocalyx thickness index was significantly lower in the isoflurane-sepsis group than propofol (P = 0.001). ConclusionsPropofol potentially offers protective action against microvascular flow deterioration compared to isoflurane, observed in control mice. Furthermore, a lower degree of sepsis-induced glycocalyx degradation was evident with propofol compared to isoflurane.

Effect of acute heparin administration on glycocalyx thickness and endothelial function in healthy younger adults.

The endothelial glycocalyx is a dynamic, gel-like layer that is critical to normal vascular endothelial function. Heparin impairs the endothelial glycocalyx and reduces vascular endothelial function in a murine model; however, this has yet to be tested in healthy humans. We hypothesized that a single bolus dose of heparin would increase circulating glycocalyx components and decrease endothelial glycocalyx thickness resulting in blunted brachial artery vasodilation in healthy younger adults. Healthy adults (n = 19, aged 18-39 yr, 53% female) underwent measurements of the endothelial glycocalyx and vascular endothelial function at baseline and after a single bolus 5,000 U dose of heparin. The glycocalyx components syndecan-1 and heparan sulfate were measured from plasma samples using enzyme-linked immunosorbent assays. Glycocalyx thickness was determined as perfused boundary region (PBR) in sublingual microvessels using the GlycoCheck. Endothelial function was measured via ultrasonography and quantified as brachial artery flow-mediated dilation (FMD). Following acute heparin administration, there was no increase in syndecan-1 or heparan sulfate (P = 0.90 and P = 0.49, respectively). In addition, there was no change in PBR 4-7 µm (P = 0.55), PBR 10-25 µm (P = 0.63), or 4-25 µm (P = 0.49) after heparin treatment. Furthermore, we did not observe a change in FMDmm (P = 0.23), FMD% (P = 0.35), or plasma nitrite concentrations (P = 0.10) in response to heparin. Finally, time to peak dilation and peak FMD normalized to shear stress were unchanged following heparin (P = 0.59 and P = 0.21, respectively). Our pilot study suggests that a single bolus intravenous dose of heparin does not result in endothelial glycocalyx degradation or vascular endothelial dysfunction in healthy younger adults.NEW & NOTEWORTHY The endothelial glycocalyx's role in modulating vascular endothelial dysfunction with aging and disease is becoming increasingly recognized. This study presents novel findings that acute heparin administration is not a feasible method to experimentally degrade the endothelial glycocalyx and measure concurrent changes in vascular endothelial function in healthy humans. Alternative approaches will be needed to translate findings from preclinical studies and test the effects of acute endothelial glycocalyx degradation on vascular endothelial function in humans.

Hydrogen attenuates endothelial glycocalyx damage associated with partial cardiopulmonary bypass in rats.

Cardiopulmonary bypass (CPB) causes systemic inflammation and endothelial glycocalyx damage. Hydrogen has anti-oxidant and anti-inflammatory properties; therefore, we hypothesized that hydrogen would alleviate endothelial glycocalyx damage caused by CPB. Twenty-eight male Sprague-Dawley rats were randomly divided into four groups (n = 7 per group), as follows: sham, control, 2% hydrogen, and 4% hydrogen. The rats were subjected to 90 minutes of partial CPB followed by 120 minutes of observation. In the hydrogen groups, hydrogen was administered via the ventilator and artificial lung during CPB, and via the ventilator for 60 minutes after CPB. After observation, blood collection, lung extraction, and perfusion fixation were performed, and the heart, lung, and brain endothelial glycocalyx thickness was measured by electron microscopy. The serum syndecan-1 concentration, a glycocalyx component, in the 4% hydrogen group (5.7 ± 4.4 pg/mL) was lower than in the control (19.5 ± 6.6 pg/mL) and 2% hydrogen (19.8 ± 5.0 pg/mL) groups (P < 0.001 for each), but it was not significantly different from the sham group (6.2 ± 4.0 pg/mL, P = 0.999). The endothelial glycocalyces of the heart and lung in the 4% hydrogen group were thicker than in the control group. The 4% hydrogen group had lower inflammatory cytokine concentrations (interleukin-1β and tumor necrosis factor-α) in serum and lung tissue, as well as a lower serum malondialdehyde concentration, than the control group. The 2% hydrogen group showed no significant difference in the serum syndecan-1 concentration compared with the control group. However, non-significant decreases in serum and lung tissue inflammatory cytokine concentrations, as well as in serum malondialdehyde concentration, were observed. Administration of 4% hydrogen via artificial and autologous lungs attenuated endothelial glycocalyx damage caused by partial CPB in rats, which might be mediated by the anti-inflammatory and anti-oxidant properties of hydrogen.

Olive extract enriched with hydroxytyrosol improves cardiac and vascular function in patients with stable coronary artery disease: a double-blind, placebo-controlled trial

Abstract Aims We investigated the effect of olive extract enriched with hydroxytyrosol, on vascular and myocardial function markers in patients with stable coronary artery disease (SCAD). Methods In a prospective, cross-over, double-blind, placebo-controlled, clinical trial, 30 patients with stable CAD were randomly assigned to an oral supplement containing olive oil extract enriched with hydroxytyrosol (OOHT) or placebo for one month and then were crossed over to the alternate treatment (placebo or OOHT for one more month). We measured a) perfused boundary region (PBR) of the sublingual arterial microvessels (increased PBR indicates reduced endothelial glycocalyx thickness), b) flow-mediated dilation (FMD), c) Coronary Flow Reserve (CFR) and markers of LV diastolic function by Doppler echocardiography d) Pulse Wave Velocity (PWV) e) oxidative/nitrosative stress and inflammatory biomarkers and blood lipids at baseline and after one month treatment. Results Treatment with OOHT improved PBR, FMD, CFR and PWV compared to baseline (1.81±0.35 vs 1.77±0.40 μm p=0.04 3.71±2.12 vs 6.51±2.31%, p&amp;lt;0.001;2.37±0.40 vs 2.45±0.41, p=0.030 and 11.03±1.81 vs 11.78±2.34 m/sec, p=0.002), while there was no effect after placebo (p&amp;gt;0.05). Compared to baseline, treatment with OOHT reduced malondialdehyde (MDA) a marker of lipid peroxidation (2.10±1.37 vs 1.69±0.83 mmol/lt, p=0.045), oxidized LDL, triglycerides, PCSK9 and CRP blood levels (p&amp;lt;0.05) in contrast to placebo. There was a parallel improvement of E’ of the mitral annulus and deceleration time of the E wave of mitral inflow after OOHT (p&amp;lt;0.05) but not after placebo. Conclusions Olive extract enriched with hydroxytyrosol improves endothelial and arterial function likely by reducing oxidative and inflammatory burden leading to improved LV diastolic function after one month of treatment in patients with stable CAD.

Association of GDF15, NTpoBNP and PLGF with subclinical cardiac dysfunction in type 2 diabetes: effects of novel antidiabetic treatment

Abstract Background In type 2 diabetics, we investigated the effects of insulin, GLP-1Ra, SGLT-2i and their combination on blood biomarkers of cardiac function and their association with left ventricular and atrial function. Patients and methods A total of 200 diabetics (60.3±10.3-year-old) treated with metformin were randomized to insulin (n= 50), liraglutide (n=50), empagliflozin (n=50) or their combination (GLP-1RA+SGLT-2i) (n= 50) . We measured at baseline and 6 months post-treatment: (a) perfused boundary region of the sublingual arterial microvessels (marker of endothelial glycocalyx thickness), (b) pulse wave velocity (PWV), (c) Left Ventricular (LV) Global Longitudinal Strain (GLS) and Torsion, (d) LV Global Work Index (GWI), Global Constructive Work (GCW), Global Wasted Work (GWW), Global Work Efficiency (GWE), (e) Left atrial reservoir strain (LASr) and (f) Growth/Differentiation Factor-15 (GDF-15), NTproBNP, Placental Growth Factor (PLGF) as markers of cardiac dysfunction. Results Increasing GGDF-15 levels at baseline were related with increasing PWV (r=0.357, p&amp;lt;0.001). Moreover, increasing GDF-15 was related with reduced absolute GLS values (r=-0.425, p=0.005), increased LV Torsion (r=0.309, p=0.035), Twisting Velocity (r=0.345, p=0.018) and E/E’ (r=0.307, p=0.041) as well as reduced Untwisting velocity (r=-0.383, p=0.008) and LASr (r=-0,388, p=0.005). Increasing NTproBNP concentration at baseline was related with increasing PWV (r=0.445, p=0.001 and PBR5-25 (r=0.222, p=0.04) . Also, increasing NTroBNP was related with reduced absolute GLS (r=-0.454, p=0.002), LV GL strain rate (r=-0.360, p=0.008) , increased LV Torsion (r=0.329, p=0.029) and E/E’ (r=0.365, p=0.021). Increasing NTproBNP was also related with decreasing GWI (r=-0.471, p&amp;lt;0.001), GWE (r=-0.555, p&amp;lt;0.001), Untwisting velocity (r=-0.383, p=0.008) and LASr (r=-0,424, p=0.002). PLGF concentration at baseline was related with reduced absolute GLS (r=-0.341, p=0.012), LV GL strain rate (r=-0.302, p=0.008) and LASr (r=-0.321, p=0.036). At 6 months, the reduction of GDF-15 and NTproBNP levels was related with the respective reduction of PWV (p=0.022 and p=0.34) Furthermore, the reduction of NTproBNP at 6 months was related with an increase of GWI and GWE (r=0.455, p=0.006/ r=0.631, p=0.001,respectively). In patients treated with GLP-1RA, SGLT-2i and their combination, there was greater improvement of GLS, Torsion, Myocardial Work, LASr, NTproBNP, GDF-15 PLGF compared to patients treated with insulin (p&amp;lt;0.05) post-treatment. In patients treated with insulin, NTproBNP, GDF-15 and PLGF changes post-treatment were not related with the respective changes of the examined cardiac and vascular markers. Conclusion Elevated levels of GDF15, PLGF and NTproBNP are associated with cardiac and vascular dysfunction in patients with type 2 diabetics and their improvement after 6 months of treatment with GLP-1RA and /or SGLT2i is associated with improved cardiovascular function.

Endothelial Glycocalyx Integrity in Treatment-Naïve People Living with HIV before and One Year after Antiretroviral Treatment Initiation.

Endothelial glycocalyx (EG) derangement has been associated with cardiovascular disease (CVD). Studies on EG integrity among people living with HIV (PLWH), are lacking. We conducted a prospective cohort study among treatment-naïve PLWH who received emtricitabine/tenofovir alafenamide, combined with either an integrase strand transfer inhibitor (INSTI, dolutegravir, raltegravir or elvitegravir/cobicistat), or a protease inhibitor (PI, darunavir/cobicistat). We assessed EG at baseline, 24 (±4) and 48 (±4) weeks, by measuring the perfused boundary region (PBR, inversely proportional to EG thickness), in sublingual microvessels. In total, 66 consecutive PLWH (60 (90.9%) males) with a median age (interquartile range, IQR) of 37 (12) years, were enrolled. In total, 40(60.6%) received INSTI-based regimens. The mean (standard deviation) PBR decreased significantly from 2.17 (0.29) μm at baseline to 2.04 (0.26) μm (p = 0.019), and then to 1.93 (0.3) μm (p < 0.0001) at 24 (±4) and 48 (±4) weeks, respectively. PBR did not differ among treatment groups. PLWH on INSTIs had a significant PBR reduction at 48 (±4) weeks. Smokers and PLWH with low levels of viremia experienced the greatest PBR reduction. This study is the first to report the benefit of antiretroviral treatment on EG improvement in treatment-naïve PLWH and depicts a potential bedside biomarker and therapeutic target for CVD in PLWH.

Effect of glucagon-like peptide-1 receptor agonists, sodium-glucose cotransporter-2 inhibitors and their combination on left atrial strain and arterial function

Abstract Funding Acknowledgements Type of funding sources: None. Background We investigated the effects of insulin, glucagon-like peptide-1 receptor agonists (GLP-1RA), sodium-glucose cotransporter-2 inhibitors (SGLT-2i) and their combination on left atrial function of patients with type 2 diabetes mellitus and we searched for a relation with arterial function. Patients and methods A total of 201 patients (59,5±9,1 year old, 161 male) with type 2 diabetes mellitus treated with metformin were randomized to insulin (n= 50 served as controls), liraglutide (n=51), empagliflozin (n=50) or their combination (GLP-1RA+SGLT- 2i) (n= 50) as add-on. We measured at baseline and 6 months posttreatment: (a) perfused boundary region of the sublingual arterial microvessels (marker of endothelial glycocalyx thickness), (b) pulse wave velocity (PWV) and central systolic blood pressure, (c) global left ventricular Longitudinal strain, (d) left atrial strain. Results Six months after intervention, all patients improved left atrial reservoir strain (GLP1RA 30,7±9,3 vs 33,9±9,7%, p = 0,011/SGLT2i 30±8,3 vs 32,3±7,3%, p = 0,04/ GLP1&amp;SGLT2i 29,1±8,7 vs 31,3±8,2, p = 0,007) in comparison with controls (33±8,3% vs 32,8±7,4, p = 0,829). Also, patients treated with GLP1RA and their combination improved left atrial conduction strain (p&amp;lt;0.05). GLP-1RA or GLP-1RA+SGLT-2i provided a greater improvement in GLS compared with insulin or SGLT-2i (controls –18,5±3,9% vs − 19,1±3%, p = 0,202/GLP1RA –18,2±4,1% vs –19,1±3,5%, p = 0,03/SGLT2i –19±4,1% vs − 19,1±3,8%, p = 0,764/GLP1&amp;SGLT2i –16,7±3,9% vs –17,5±4,3%, p = 0,026). SGLT-2i or GLP-1RA and SGLT-2i showed a greater decrease of PWV (Controls 12,06±3,02 vs 11,62±2,79 m/sec, p = 0,142/ GLP1 11,54±2,91 vs 11,45±2,52 m/sec, p = 0,822/ SGLT2 11,86±2,51 vs 11,33±2,28 m/sec, p = 0,047/ GLP1&amp;SGLT2 12,01±2,46 vs 10,85±1,79 m/sec, p = 0,021) and central and brachial systolic blood pressure than insulin or GLP-1RA. At baseline, there was a correlation of LA reservoir strain with PWV (r =−0,209, p = 0,008), central SBP (r =−0,151, p = 0,030), EF (r=0,214, p = 0,004), GLS (r =−0,279, p = 0,009). At 6 months follow up, there was a correlation of LA reservoir strain with PWV (r=0,217, p = 0,003), central SBP (r =0,214, p = 0,004), EF (r=–0,221, p = 0,049), GLS (r =0,275, p = 0,014). Conclusions Treatment with GLP-1RA, SGLT-2i, and their combination for 6 months showed a greater improvement of left atrial function compared to insulin treatment in parallel with improvement arterial and myocardial function.

Impaired endothelial glycocalyx, vascular dysfunction and myocardial deformation four months after Covid-19 infection are partially improved at twelve months

Abstract Funding Acknowledgements Type of funding sources: None. Introduction COVID-19 patients present impaired subclinical markers of cardiovascular and endothelial function. Subclinical myocardial and vascular dysfunction during COVID-19 disease have been associated with worse outcomes and higher mortality risk. Purpose We investigated the effect of COVID-19 infection on markers of endothelial, vascular and myocardial function at four and twelve months after the infection Methods We recruited 70 patients who were examined in a dedicated post-COVID-19 outpatient clinic during a scheduled follow-up visit at four and twelve months after a confirmed COVID-19 infection and 70 healthy individuals with similar clinical characteristics. At four and twelve months we measured (i) perfused boundary region (PBR) of the sublingual arterial microvessels (increased PBR indicates reduced endothelial glycocalyx thickness), (ii) flow-mediated dilatation (FMD), (iii) coronary flow reserve (CFR) by Doppler echocardiography, (iv) pulse wave velocity (PWV) and central systolic blood pressure (cSBP), (v) global left and right ventricular longitudinal strain (GLS), (vi) myocardial global work index (GWI) global constructive work (GCW), global wasted work (GWW) and the myocardial global work efficiency (GWE) and v) malondialdehyde (MDA), an oxidative stress marker. Results At four months, COVID-19 patients displayed higher values of PBR5–25 compared to control group (p&amp;lt;0.001) which increased at twelve months (p&amp;lt;0.001). FMD, PWV and cSBP values were similar between 4 and 12 months (p&amp;gt;0.05 for all the comparisons) and higher than those in controls (p&amp;lt;0.001, p = 0.057, p = 0.003 respectively). At four months, COVID-19 patients presented impaired CFR and LVGLS values which were improved at twelve months (p = 0.002, p = 0.069 respectively), though remained impaired compared to controls (p = 0.003 for all the comparisons). At four months, COVID-19 patients had impaired RVGLS values which were significantly improved at twelve months (p = 0.001) and showed no statistically significant difference compared to controls (p&amp;gt;0.05). COVID-19 patients at four months display higher myocardial wasted work and decreased myocardial efficiency compared to controls (p = 0.01, p = 0.006, respectively). There was a modest improvement in GWW and GWE at twelve months, (p = 0.043, p = 0.001, respectively); however, these markers remained impaired compared to controls (p&amp;gt;0.05). At four months, MDA was higher in COVID-19 patients compared to control group and significantly decreased at twelve months (p&amp;lt;0.001); however, these values remain higher than in controls (p = 0.002). Conclusions SARS-CoV-2 causes endothelial and cardiovascular dysfunction which are partially restored at twelve months after the infection.

Olive extract enriched with hydroxytyrosol improves cardiac and vascular function in patients with stable coronary artery disease: a double-blind, placebo-controlled trial

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – EU funding. Main funding source(s): Olive Heart Operational Programme Competitiveness, Entrepreneurship and Innovation 2014–2020 (EPAnEK) Aims We investigated the effect of olive extract enriched with hydroxytyrosol, on vascular and myocardial function markers in patients with stable coronary artery disease (SCAD). Methods In a prospective, cross-over, double-blind, placebo-controlled, clinical trial, 30 patients with stable CAD were randomly assigned to an oral supplement containing olive oil extract enriched with hydroxytyrosol (OOHT) or placebo for one month and then were crossed over to the alternate treatment (placebo or OOHT for one more month). We measured a) perfused boundary region (PBR) of the sublingual arterial microvessels (increased PBR indicates reduced endothelial glycocalyx thickness), b) flow-mediated dilation (FMD), c) Coronary Flow Reserve (CFR) and markers of LV diastolic function by Doppler echocardiography d) Pulse Wave Velocity (PWV) e) oxidative/nitrosative stress and inflammatory biomarkers and blood lipids at baseline and after one month treatment. Results Treatment with OOHT improved PBR, FMD, CFR and PWV compared to baseline (1.81±0.35 vs 1.77±0.40 μm p = 0.04 3.71±2.12 vs 6.51±2.31%, p&amp;lt;0.001;2.37±0.40 vs 2.45±0.41, p = 0.030 and 11.03±1.81 vs 11.78±2.34 m/sec, p = 0.002), while there was no effect after placebo (p&amp;gt;0.05). Compared to baseline, treatment with OOHT reduced malondialdehyde (MDA) a marker of lipid peroxidation (2.10±1.37 vs 1.69±0.83 mmol/lt, p = 0.045), oxidized LDL, triglycerides, PCSK9 and CRP blood levels (p&amp;lt;0.05) in contrast to placebo. There was a parallel improvement of E’ of the mitral annulus and deceleration time of the E wave of mitral inflow after OOHT (p&amp;lt;0.05) but not after placebo. Conclusions Olive extract enriched with hydroxytyrosol improves endothelial and arterial function likely by reducing oxidative and inflammatory burden leading to improved LV diastolic function after one month of treatment in patients with stable CAD.

The counterbalance of endothelial glycocalyx and high wall shear stress to low-density lipoprotein concentration polarization in mouse ear skin arterioles

Background and aimsAtherosclerosis preferentially occurs at regions in arterial branching, curvature, and stenosis, which may be explained by the geometric predilection of low-density lipoprotein (LDL) concentration polarization that has been investigated in major arteries in previous studies. Whether this also happens in arterioles remains unknown. MethodsHerein, a radially non-uniform distribution of LDL particles and a heterogeneous endothelial glycocalyx layer in the mouse ear arterioles, as shown by fluorescein isothiocyanate labeled wheat germ agglutinin (WGA-FITC), were successfully observed by a non-invasive two-photon laser-scanning microscopy (TPLSM) technique. The stagnant film theory was applied as the fitting function to evaluate LDL concentration polarization in arterioles. ResultsThe concentration polarization rate (CPR, the ratio of the number of polarized cases to that of total cases) in the inner walls of curved and branched arterioles was 22% and 31% higher than the outer counterparts, respectively. Results from the binary logistic regression and multiple linear regression analysis showed that endothelial glycocalyx thickness increases CPR and the thickness of the concentration polarization layer (CPL). Flow field computation indicates no obvious disturbances or vortex in modeled arterioles with different geometries and the mean wall shear stress is about 7.7–9.0 Pa. ConclusionsThese findings suggest a geometric predilection of LDL concentration polarization in arterioles for the first time, and the existence of an endothelial glycocalyx, acting together with a relatively high wall shear stress in arterioles, may explain to some extent why atherosclerosis rarely occurs in these regions.

Relationship of the main indicators of systemic COVID-associated endotheliopathy with the morphofunctional state and hemodynamics of the retina and chorioid in the acute period of the disease

BACKGROUND: Nonspecific angio- and retinopathy is one of the clinical manifestations of a new coronavirus infection. The frequency of occurrence of these changes in people with severe COVID-19 does not exceed 55%. The causes, course and consequences of these microcirculatory disorders of the retina are currently not well understood.&#x0D; AIM: To study and compare of retinal morphometric parameters and systemic endothelial dysfunction markers, as well as the main clinical and laboratory parameters in patients with moderate and severe coronavirus infection during convalescence.&#x0D; MATERIALS AND METHODS: The study involved 44 patients (86 eyes) who had COVID-19 during the previous 3 months, who were divided into 2 groups: with moderate and severe disease. The control group consisted of 18 healthy volunteers (36 eyes). All patients underwent a standard ophthalmological examination and optical coherence tomography, which included an assessment of the choroidal thickness (CT) and measurement of the mean diameter of the peripapillary arteries (MAD) and veins (MVD). During hospitalization, all patients underwent a laboratory study of venous blood parameters, as well as an assessment of the microcirculation of the sublingual plexus by examining the density of the endothelial glycocalyx (PBR) using the GlycoCheck.&#x0D; RESULTS: In patients who underwent COVID-19, there was a significant increase in CT relative to the control group, amounting to 308, 344 and 392 m, respectively. The most pronounced difference was observed between MVD in patients with severe infection and the control group (119.1 m vs. 99.2 m). In patients with moderate and severe COVID-19, MAD and MVD were positively correlated with TC, with r = 0.389 and r = 0.584, respectively. MVD also correlated with the level of leukocytes (r = 0.504), the ESR value (r = 0.656). Correlations between MVD and data characterizing the state of the glycocalyx in the sublingual vascular plexus were revealed: the filling of small capillaries with erythrocytes (r = 0.587), as well as the marginal perfusion value in large capillaries 2025 m (r = 0.479) and PBR (r = 0.479). Only significant differences and correlations are shown (p 0.005).&#x0D; CONCLUSIONS: In patients who underwent moderate and severe COVID-19 during the convalescence period (up to 30 days), an increase in the diameter of peripapillary vessels and TC is observed, proportional to the severity of COVID-19, laboratory markers of systemic inflammation and hypercoagulation (the number of leukocytes, the ESR value, D-dimer and prothrombin), which indicates the inflammatory nature of the changes. The severity of postcovid retinal microangiopathy correlates with indicators detecting a decreasing of the endothelial glycocalyx thickness in the sublingual capillary plexus, which indirectly indicates a connection with systemic endotheliopathy.

Reduction of Endothelial Glycocalyx on Peritubular Capillaries in Chronic Kidney Disease

In chronic kidney disease (CKD), peritubular capillaries undergo anatomic and functional alterations, such as rarefaction and increased permeability. The endothelial glycocalyx (EG) is a carbohydrate-rich gel-like mesh, which covers the luminal surface of endothelial cells. It is involved in many regulatory functions of the endothelium, including vascular permeability. Herein, we investigated ultrastructural alterations of the EG in different murine CKD models. Fluorescence staining using different lectins with high affinity to components of the renal glycocalyx revealed a reduced binding to the endothelium in CKD in the animal models, and there were similar finding in human kidney specimens. We used the Lanthanum Dysprosium Glycosamino Glycan adhesion staining technique to visualize the ultrastructure of the glycocalyx in transmission electron microscopy. This also enabled quantitative analyses, showing a significant reduction of the EG thickness and density. In addition, mRNA expression of proteins involved in glycocalyx biology, synthesis, and turnover (ie, syndecan 1 and glypican 1), which are main components of the glycocalyx, and exostosin 2, involved in the synthesis of the glycocalyx, were significantly up-regulated in endothelial cells isolated from murine CKD models. Visualization of glycocalyx using specific transmission electron microscopy analyses allows qualitative and quantitative analyses and revealed significant pathologic alterations in peritubular capillaries in CKD.

Four-months treatment with dapagliflozin improves endothelial glycocalyx and cardiovascular function in patients with type 1 diabetes mellitus

Abstract Background/Introduction Patients with type 1 diabetes mellitus (T1DM) present signs of vascular and endothelial dysfunction earlier compared to healthy individuals. According to clinical studies, SGLT-2i favorably affect cardiovascular outcomes in patients with type 2 diabetes mellitus (T2DM). However, the data regarding the efficacy of SGLT-2i on cardiovascular markers in patients with T1DM are scarce. Purpose We investigated the effects of dapagliflozin on endothelial and cardiovascular function in patients with T1DM. Methods We recruited in total 30 patients with T1DM and pour glycemic control who were under medication with insulin and received dapagliflozin (n=15) or intensification of insulin treatment (n=15) (control group). We measured at baseline and at four months post-treatment the: a) Carotid-femoral pulse wave velocity (PWV-Complior; ALAM Medical) b) Central systolic blood pressure (cSBP) c) Perfused boundary region (PBR) of the sublingual arterial microvessels (marker of endothelial glycocalyx thickness) and d) Left ventricular global longitudinal strain (GLS) using speckle-tracking echocardiography. Results At baseline, patients among the two groups had similar age, sex, HbA1c and markers of endothelial and cardiovascular function (p&amp;gt;0.05). After four months treatment, patients who received dapagliflozin displayed an improvement in PBR5–25 (−41%, p&amp;lt;0.05), in PWV (−10%, p&amp;lt;0.05), in cSBP (−6%, p&amp;lt;0.05) and in GLS (+5%, P&amp;lt;0.05) compared to baseline. However, no statistically significant changes in PBR5–25, in PWV, in cSBP and in GLS were observed after intensification of insulin treatment (PBR5–25: +3%, PWV: +2%, cSBP: −2%, GLS: −3% at 4 months, P&amp;gt;0.05), despite a similar HbA1c reduction (Table 1). Changes of PBR after four months treatment with dapagliflozin correlated with a concomitant reduction of PWV and cSBP (P&amp;lt;0.05). Conclusions Four-months treatment with dapagliflozin improves endothelial glycocalyx and cardiovascular function in patients with T1DM, independently of glycemic control. Funding Acknowledgement Type of funding sources: None.

Four-month treatment with long-acting glucagon-like peptide-1 receptor agonists (GLP-1) or sodium-glucose co-trasporter-2 inhibitors (SGLT-2i) improves vascular function in type 2 diabetes mellitus

Abstract Background/Introduction Patients with type 2 diabetes mellitus (T2DM) and ischemic stroke present impaired markers of vascular and endothelial function. Glucagon-like peptide-1 receptor agonists (GLP-1) and sodium-glucose contrasporter-2 inhibitors (SGLT-2) are novel antidiabetic agents reducing the risk of cardiovascular complications. Purpose The aim of the study is to investigate the effect of treatment with GLP-1 or SGLT2 on arterial stiffness and endothelial glycocalyx in patients with T2DM and ischemic stroke. Methods We recruited in total 81 patients with T2DM and ischemic stroke who received dulaglutide (n=27), dapagliflozin (n=27) or insulin (n=27). We measured at baseline and at four months post-treatment the: a) Carotid-femoral pulse wave velocity (PWV-Complior; ALAM Medical), b) Augmentation index (Aix), c) Central systolic blood pressure (cSBP), and d) Perfused boundary region (PBR) of the sublingual arterial microvessels(marker of endothelial glycocalyx thickness), e) Left ventricular global longitudinal strain (GLS) using speckle-tracking echocardiography. Results At baseline, patients among the three groups had similar age, sex, HbA1c and markers of endothelial and cardiovascular function (p&amp;gt;0.05). Dapagliflozin and dulaglutide showed a greater reduction of PBR, PWV and central SBP than insulin, despite a similar HbA1c reduction (p&amp;lt;0.05). After four months treatment, patients on dapagliflozin and on dulaglutide displayed a greater reduction of PWV (−10% and −8% vs −1%), of cSBP (−9% and −8% vs +1%), of Aix (−65% and −13% vs −3%) and of GLS (+11% and +7% vs +5%) compared to patients on insulin (p&amp;lt;0.05 for all comparisons) (Table 1). PBR values were improved only in patients who received dulaglutide (−4% vs dapagliflozin: −2% vs insulin +1%). Conclusions Dulaglutide and dapagliflozin improve cardiovascular function, but only dulaglutide improves endothelial glycocalyx in patients with T2DM and ischemic stroke after four months treatment. Funding Acknowledgement Type of funding sources: None.

Impaired endothelial glycocalyx, vascular dysfunction and myocardial deformation four months after COVID-19 infection are partially improved at twelve months

Abstract Introduction COVID-19 patients present impaired subclinical markers of cardiovascular and endothelial function. Subclinical myocardial and vascular dysfunction during COVID-19 disease have been associated with worse outcomes and higher mortality risk. Purpose We investigated the effect of COVID-19 infection on markers of endothelial, vascular and myocardial function at four and twelve months after the infection Methods We recruited 70 patients who were examined in a dedicated post-COVID-19 outpatient clinic during a scheduled follow-up visit at four and twelve months after a confirmed COVID-19 infection and 70 healthy individuals with similar clinical characteristics. At four and twelve months we measured (i) perfused boundary region (PBR) of the sublingual arterial microvessels (increased PBR indicates reduced endothelial glycocalyx thickness), (ii) flow-mediated dilatation (FMD), (iii) coronary flow reserve (CFR) by Doppler echocardiography, (iv) pulse wave velocity (PWV) and central systolic blood pressure (cSBP), (v) global left and right ventricular longitudinal strain (GLS), (vi) myocardial global work index (GWI) global constructive work (GCW), global wasted work (GWW) and the myocardial global work efficiency (GWE) and v) malondialdehyde (MDA), an oxidative stress marker. Results At four months, COVID-19 patients displayed higher values of PBR5–25 compared to control group (p&amp;lt;0.001) which increased at twelve months (p&amp;lt;0.001). FMD, PWV and cSBP values were similar between 4 and 12 months (p&amp;gt;0.05 for all the comparisons) and higher than those in controls (p&amp;lt;0.001, p=0.057, p=0.003 respectively). At four months, COVID-19 patients presented impaired CFR and LVGLS values which were improved at twelve months (p=0.002, p=0.069 respectively), though remained impaired compared to controls (p=0.003 for all the comparisons). At four months, COVID-19 patients had impaired RVGLS values which were significantly improved at twelve months (p=0.001,) and showed no statistically significant difference compared to controls (p&amp;gt;0.05). COVID-19 patients at four months display higher myocardial wasted work and decreased myocardial efficiency compared to controls (p=0.01, p=0.006 respectively). There was a modest improvement in GWW and GWE at twelve months,(p=0.043, p=0.001, respectively); however, these markers remained impaired compared to controls (p&amp;gt;0.05). At four months, MDA was higher in COVID-19 patients compared to control group and significantly decreased at twelve months (p&amp;lt;0.001); however, these values remain higher than in controls (p=0.002) (Table 1). Conclusions SARS-CoV-2 causes endothelial and cardiovascular dysfunction which are partially restored at twelve months after the infection. Funding Acknowledgement Type of funding sources: None.

SARS-CoV-2 is associated withabnormal biomarkers of oxidative stress,and endothelial function linked with cardiovascular dysfunction four months after the infection

Abstract Introduction COVID-19 infection has been associated with increase arterial stiffness, endothelialdysfunction, and impairment in coronary and cardiac performance. Inflammation and oxidative stress have beensuggested as possible pathophysiological mechanisms leading to vascular and endothelial deregulation afterCOVID-19 infection. Purpose The objective of our study is to evaluate premature alterations in arterial stiffness, endothelial,coronary, and myocardial function markers four months after SARS-CoV-2 infection. Methods In a case-control prospective study, we included 70 patients 4 months after COVID-19 infection, 70 age- and sex-matched untreated hypertensive patients (positive control) and 70 healthy individuals. We measured (i) perfused boundary region (PBR) of the sublingual arterial microvessels (increased PBR indicates reduced endothelial glycocalyx thickness), (ii) flow-mediated dilatation (FMD), (iii) coronary flow reserve (CFR) by Doppler echocardiography, (iv) pulse wave velocity (PWV) and central systolic blood pressure (cSBP), (v) global left and right ventricular longitudinal strain (GLS), (vi) malondialdehyde (MDA), an oxidative stress marker, thrombomodulin and von Willebrand factor as endothelial biomarkers. Results COVID-19 patients had similar CFR and FMD with hypertensives (2.48±0.41 vs 2.58±0.88, p=0.562, 5.86±2.82% vs 5.80±2.07%, p=0.872 respectively) but lower values than controls (3.42±0.65, p=0.0135, 9.06±2.11%, p=0.002 respectively). Compared to controls, both COVID-19 and hypertensives had greater PBR5–25 (2.07±0.15μm and 2.07±0.26μm p=0.8 vs 1.89±0.17μm, p=0.001), higher PWV, (12.09±2.50 vs 11.92±2.94, p=0.7 vs 10.04±1.80m/sec, p=0.036) increased cSBP (128.43±17.39 vs 135.17±16.83 vs 117.89±18.85) and impaired LV and RV GLS (−19.50±2.56% vs −19.23±2.67%, p=0.864 vs −21.98±1.51%, p=0.020 and −16.99±3.17% vs −18.63±3.20%, p=0.002 vs −20.51±2.28%, p&amp;lt;0.001). MDA and thrombomodulin were higher in COVID-19 patients than both hypertensives and controls (10.67±2.75 vs 1.76±0.30, p=0.003 vs 1.01±0.50nmole/L, p=0.001 and 3716.63±188.36 vs 3114.46±179.18, p=0.017 vs 2590.02±156.51pg/ml, p&amp;lt;0.001). COVID-19 patients displayed similar vWF values with hypertensives but higher compared with healthy controls (4018.03±474.31 vs 3756.65±293.28 vs 2079.33±855.10 ng/ml, p=0.718 and p=0.016 respectively). Conclusions SARS-CoV-2 infection is associated with oxidative stress, endothelial and vascular dysfunction, which are linked to impaired longitudinal myocardial deformation 4 months after COVID-19 infection. Funding Acknowledgement Type of funding sources: None.

Endothelial glycocalyx integrity in oncological patients

BackgroundCancer is associated with early changes in the cardiovascular system (CV) before overt cardiotoxicity. Endothelial dysfunction is induced by chemotherapeutic regimens but there is no data for endothelial glycocalyx in cancer. MethodsSixty-four patients with cancer (65.6% with solid tumors and 34.4% with hematological malignancies) and 32 controls from the outpatient cardiology clinic were included in the study. The perfused boundary region (PBR) of the sublingual arterial microvessels, Pulse Wave Velocity (PWV) and augmentation index (AI) were measured. A standard transthoracic echocardiogram plus assessment of global longitudinal strain (GLS) of all cardiac chambers were performed. ResultsThere was no difference in the baseline profile (age, sex, smoking, hypertension, diabetes, hyperlipidemia and coronary artery disease) and in the echocardiographic parameters between the two groups, with the exception of left atrial volume (33.3 ± 13 in cancer patients vs 27.6 ± 6.5 ml/m2 in controls). PBR 5–25 and PBR 20–25 were significantly increased in cancer patients vs controls (2.11 ± 0.36 vs 1.97 ± 0.21 μm, p = 0.025 and 2.65 ± 0.48 vs 2.40 ± 0.36 μm, p = 0.012, respectively). Endothelial glycocalyx thickness impairment was independent of traditional CV risk factors and anticancer therapy, but proportional to disease stage (r = 0.337, p = 0.044). However, there was no difference in arterial stiffness between the two groups (PWV 10.74 ± 4.11 vs 11.26 ± 3.38 m/s, p = 0.539 and AI 11.28 ± 28.87 vs 15.38 ± 18.8 %, p = 0.470). ConclusionsEndothelial function as assessed by endothelial glycocalyx thickness is significantly impaired in cancer patients without overt cardiotoxicity. This implies that PBR might be useful for the early assessment of microvascular and endothelial toxicity of cancer.