Migraine is a neurological disorder with recurrent attacks characterized by headaches and sensitivity to stimuli. Stress is a significant trigger for attacks; however, molecular mechanisms of the connection are poorly understood.

To better characterize such mechanisms, we performed a placebo-controlled, double-blind crossover study with 51 participants (21 patients with migraine without aura and 30 healthy controls).

Participants received a low-dose citalopram- or placebo challenge on 2 separate days. Prechallenge and postchallenge assessment of cortisol concentrations and transcriptomic changes using RNA-seq was performed from whole blood samples. Analysis of an accidental attack following the citalopram challenge was also conducted.

Neuroendocrine challenge elicited elevated cortisol concentrations at 30 (P-value = 0.1355) and 70 minutes (P-value = 0.07292) postchallenge in patients with migraine compared with controls. Gene expression analysis showed 10 differentially expressed genes (2 down- and 8 upregulated, P-value ≤ 0.005) and 10 dysregulated gene sets (P-value ≤ 0.005). Among them, dysregulated IKBKGP1 and NKRF genes and upregulated protein synthesis and translation, carbohydrate metabolism, and, attack-related, glycosylation can be highlighted.

Patients with migraine without aura showed an enhanced cortisol response to a neuroendocrine challenge. This was accompanied by a probable suppression of NFκB activity through dysregulation of NKRF and an altered immune function. Upregulated carbohydrate metabolism may reflect the elevated cortisol concentrations' stimulating effects on endothelial glycocalyx, playing a role in NO-induced vasodilation, a trigger for migraine attacks. The results suggest the elevated cortisol response may trigger migraine attacks through altered glycocalyx and immune functions.

Sodium glucose cotransporter 2 inhibitors lower blood pressure. The underlying mechanisms are multifactorial and include effects on vascular function. We examined the systemic hemodynamic effects of empagliflozin in patients with type 2 diabetes mellitus (DM2) with and without chronic kidney disease (CKD) and in patients with nondiabetic CKD.

Three double-blinded, randomized, placebo-controlled cross-over trials, including patients with DM2 and preserved renal function ( n  = 16), DM2 and CKD ( n  = 17) and nondiabetic CKD ( n  = 16). Participants were randomized to 4 weeks of empagliflozin 10 mg or placebo and crossed over after a 2-week washout. We measured brachial and central 24-h ambulatory blood pressure (ABP), pulse wave velocity (PWV), augmentation index (AIx@75), markers of nitric oxide and erythrocyte sodium sensitivity (ESS), a marker of endothelial glycocalyx function.

Empagliflozin reduced PWV [-0.16 m/s, 95% confidence interval (95% CI): -0.26; -0.06, P  = 0.002], AIx@75 (-2.17%, 95% CI: -3.31; -1.02, P  < 0.001) and brachial and central ABP in the combined study population ( n  = 49). Changes in PWV and AIx@75 correlated to changes in systolic brachial ABP. Markers of nitric oxide did not increase, but empagliflozin decreased ESS, which was correlated to an increase in haematocrit.

Empagliflozin decreased arterial stiffness, mediated partly by a decrease in brachial ABP. We found no increase in nitric oxide activity, but ESS decreased. While this may be explained partly by a change in haematocrit, it could indicate an improvement in endothelial glycocalyx function.

EU Clinical Trials Register 2019-004303-12, 2019-004447-80 and 2019-004467-50.

The endothelial glycocalyx is a dynamic brush-like layer composed of proteoglycans and glycosaminoglycans, including heparan sulfate (HS) and hyaluronic acid (HA), and is an important regulator of vascular homeostasis. Its structure (thickness ranges from 20 to 6450 nm in different species) not only provides a charge-selective barrier but also serves to anchor mechanosensors such as the glypican-1 (GPC-1)/caveolin-1 (CAV-1) complex and buffers shear stress. In severe acute pancreatitis (SAP), inflammatory factors promote the expression of matrix metalloproteinases (MMPs) and heparinases, which degrade syndecan-1 (SDC-1) and HS, while oxidative stress disrupts HA-CD44 binding, leading to increased capillary leakage and neutrophil adhesion. This degradation process occurs before the onset of multiple organ dysfunction syndrome (MODS), highlighting the potential of the glycocalyx as an early biomarker. More importantly, the regeneration of glycocalyx through endothelial cell synthesis of glycosaminoglycans (GAGs) and shear stress-driven SDC recycling provides therapeutic prospects. This review redefines the pathophysiology of severe acute pancreatitis-associated multiple organ dysfunction (SAP-MODS) by exploring the glycocalyx's central mechanistic role and proposes stabilizing glycocalyx structure as a potential strategy to prevent microcirculatory failure.

The pericellular matrix (PCM) is the immediate microniche surrounding cells in various tissues, regulating matrix turnover, cell-matrix interactions, and disease. This study elucidates the structure-mechanical properties and mechanobiology of the PCM in fibrocartilage, using the murine meniscus as the model. The fibrocartilage PCM is comprised of thin, randomly oriented collagen fibrils that entrap proteoglycans, contrasting with the densely packed, highly aligned collagen fibers in the bulk extracellular matrix (ECM). Compared to the ECM, the PCM exhibits lower modulus and greater isotropy, but has similar relative viscoelastic properties. In Col5a1+/- menisci, the reduction of collagen V results in thicker, more heterogeneous collagen fibrils, reduced modulus, loss of isotropy and faster viscoelastic relaxation in the PCM. Such altered PCM leads to impaired matrix-to-cell strain transmission, and in turn, disrupts mechanotransduction of meniscal cells, as illustrated by reduced calcium signaling activities and alters expression of matrix genes. In vitro, Col5a1+/- cells produce a weakened PCM with inferior properties and reduced protection of cells against tensile stretch. These findings highlight the PCM as a distinctive microstructure in fibrocartilage mechanobiology, underscoring a pivotal role of collagen V in PCM function. Targeting the PCM or its constituents offers potential for improving meniscus regeneration, osteoarthritis intervention and broader fibrocartilage-related therapies.

Background: Glycocalyx disintegration is associated with adverse outcomes in patients with trauma or sepsis. As microvascular dysfunction has an impact on disease progression in chronic heart failure (CHF) patients, we hypothesized that changes in microcirculation might be associated with mortality. Methods: Fifty patients with ischemic and non-ischemic cardiomyopathy and conservative treatment with baseline measurements of the sublingual microcirculation (via Sidestream Darkfield videomicroscopy) were followed up for two years. Glycocalyx thickness was assessed indirectly by calculation of the perfused boundary region (PBR). Results: Loss of glycocalyx was pronounced in non-survivors after one, n = 10, and two years, n = 16; PBR: 2.05 μm (1.88-2.15 μm) vs. 1.87 μm (1.66-2.03 μm) and 2.04 (1.93-2.11) vs. 1.84 (1.62-1.97); p = 0.042 and p = 0.003, respectively. Area under the ROC curve for the analysis of the predictive value of PBR on two-year mortality was 0.77 (p = 0.003; SE: 0.07, CI (95%): 0.63-0.91). ROC curve analysis determined a PBR of 1.9 μm as the best predictor for two-year mortality (sensitivity: 0.81; specificity: 0.59). Moreover, multivariate regression analysis revealed PBR and functional capillary density as significant predictors of two-year mortality, p = 0.036 and p = 0.048, respectively. Conclusions: Glycocalyx disintegration is related to poor overall survival in CHF patients.

ObjectivesLower leg edema is common in the late afternoon even in healthy individuals and could involve venous blood flows. Women appear more likely to develop lower leg edema, possibly due to the menstrual cycle. However, this phenomenon has not been quantitatively investigated using imaging. This study therefore used sonography to investigate sex-dependent impacts on physiological lower leg edema, in relation to venous blood flows in the legs and the menstrual cycle.MethodsParticipants comprised 54 healthy young adults (30 men, 24 women; mean age, 21 ± 1 years). Thickness of the papillary dermis and thickness and echogenicity of the subcutaneous adipose tissue were measured in the lower leg. Popliteal vein hemodynamics were investigated using Doppler sonography. Values were compared between late afternoon and morning. Each comparison was performed for men, women in the follicular, and women in the luteal phase.ResultsFor women in the luteal phase, papillary dermis and subcutaneous adipose tissue were thicker (median 0.20 mm, interquartile range [IQR] 0.12-0.25 mm vs median 0.33 mm, IQR 0.25-0.35 mm; p < .001 and median 5.0 mm, IQR 4.3-5.5 mm vs median 5.2 mm, IQR 4.5-6.2 mm; p = .013, respectively) and subcutaneous adipose tissue echogenicity was higher (median 66.9 IQR 64.1-70.5 vs median 71.7, IQR 65.0-76.7; p = .007) in the late afternoon than in the morning. The popliteal vein velocity-time integral (VTI) was lower in the late afternoon (median 33.0 cm, IQR 27.3-40.5 cm) than in the morning (median 26.1 cm, IQR 23.5-39.6 cm; p = .043). A significant negative correlation was observed between VTI and echogenicity (r = -0.549, p = .005). These findings were reduced in women in the follicular phase, and absent in men.ConclusionLate-afternoon lower leg edema may be associated with decreased leg blood flow in women, particularly in the luteal phase.

Intravenous fluid therapy is a regular component of care for cats and dogs with goals of restoring tissue perfusion, preventing organ dysfunction, and maintaining homeostasis. Veterinarians have a plethora of fluid choices, including hypertonic, hypotonic, and isotonic crystalloids, as well as synthetic and natural colloids. Studies published in both the human and veterinary medical literature have documented adverse events associated with synthetic and natural colloids. Reports of increased incidences of acute kidney injury, need for renal replacement therapy, and mortality in a variety of patient populations have called into question the use of synthetic colloids in humans and companion animals.

The endothelial glycocalyx is an anti-inflammatory, anti-coagulant coating of endothelial cells which participates in many physiologic processes. Damage to the glycocalyx occurs secondary to several disease states, and is especially pronounced in ischemia-reperfusion (I/R) injury. In models of trauma and hemorrhage, this damage has been shown to occur secondary to disordered succinate metabolism leading to increased mitochondrial reactive oxygen species (mitoROS). Aortic occlusion has also been associated with I/R injury to the vascular endothelium, though the precise mechanisms by which this occurs, and whether that injury includes glycocalyx shedding, are largely unknown. Consequently, very few therapeutic strategies for protection of the glycocalyx have been investigated. The aim of this study was to determine whether an experimental model of infrarenal aortic occlusion, with or without hemorrhage, causes a mitochondrial ROS-dependent glycocalyx shedding, and at what point this shedding predominantly occurs. We additionally aimed to determine whether plasma succinate was elevated following aortic occlusion.

Male Sprague Dawley rats were anesthetized and laparotomy performed. A jugular catheter was placed and a baseline blood sample collected. In the hemorrhage group, 3 mL of blood was drawn from the jugular catheter. The infrarenal abdominal aorta was isolated and clamped with an atraumatic vascular clamp. Thirty minutes after clamping a second blood sample was collected, followed by unclamping. Blood samples were collected 2 and 15 min after reperfusion. Blood was centrifuged, and succinate and glycocalyx-component syndecan-1 were measured in the plasma using an ELISA. All rats in the treatment arm received identical hemorrhage, clamping, and blood sampling, but were treated with mitochondrial-targeted ROS scavenger mitoTEMPOL prior to clamping the aorta. After euthanizing the rats, calf muscle of the hemorrhage group was flash frozen, sectioned, and stained for glycocalyx with wheat germ agglutinin.

Plasma syndecan-1 levels were significantly (P < 0.05) elevated 2 mins after unclamping the aorta compared to baseline in both groups. Levels remained increased, though not statistically significantly, at 15 min after reperfusion. No significant increase in syndecan-1 was seen after 30 min of ischemia. MitoTEMPOL treatment prevented an increase in plasma syndecan-1. Correspondingly, glycocalyx staining intensity in mitoTEMPOL-treated rats was increased compared to control. Plasma succinate was significantly elevated at 15 min after reperfusion and was not affected by treatment with mitoTEMPOL.

To our knowledge this is the first study which demonstrates a successful therapeutic strategy in the treatment of endothelial glycocalyx shedding caused by aortic occlusion and hemorrhage in an animal model. Our study shows that glycocalyx shedding is attributable to mitochondrial ROS, and occurs following distal reperfusion rather than occlusion of the aorta or during hemorrhage. Consistent with findings in hemorrhagic shock, plasma succinate levels are elevated as a result of aortic occlusion and are upstream of ROS. Taken together, our findings suggest that in the setting of aortic occlusion, a therapeutic intervention could be performed prior to reperfusion which would substantially reduce injury to the vascular endothelium.

Knowledge of the relative importance of advection and diffusion in clearing waste from the brain has been elusive, especially concerning the extracellular space (ECS). With local and global computational models of the mouse brain, we explore how the presence or absence of advection in the ECS affects solute transport. Without advection in the ECS, clearance would occur by diffusion into flowing cerebrospinal fluid in perivascular spaces (PVSs) or elsewhere, but we find this process to be severely limited by build-up of solute in the PVSs. We simulate flow in the ECS driven by a pressure drop between arteriole and venule PVSs, which enhances clearance considerably. To assess the relative importance of advection and diffusion, we introduce a local Péclet number [Formula: see text], a dimensionless scalar field. For our simulations, [Formula: see text] through much of the ECS but [Formula: see text] near PVSs near the brain surface. This local dominance of advection in the ECS establishes a clearance mechanism markedly different from that produced by diffusion alone. In network simulations that explore different parameter values and efflux routes, the pressures needed to drive the PVS flows measured in vivo are unrealistically large for most cases lacking ECS flow. Collectively, our models indicate that a flow in the ECS is necessary to explain experimental measurements and maintain homeostasis.

The use of neural tracers as targeting molecules for drug delivery has been previously established as a novel and efficient method of neural drug delivery. The wheat germ agglutinin-horseradish peroxidase conjugate (WGAHRP) is a common neural tracer, which has been used extensively to decipher neural pathways in vertebrates. It has been widely reported to bind to cell surfaces and be transported in a retrograde fashion (from synapses toward the cell body) via dynein motors along microtubules within axons and transynaptically between neurons. Here we report on the differential binding between WGAHRP and gold-conjugated WGAHRP (AuNP-WGAHRP) to the glycoprotein profiles extracted from two neuronal cell lines and one skeletal muscle cell line, as well as the binding kinetics to heparin. From proteomic analysis of the extracted glycoproteins, we suggest the identity of cell surface glycoproteins involved in the retrograde transport of WGAHRP. This study illuminates the interfacial and molecular interactions of protein-gold conjugates with native ligands and opens the door for the identification of new targets for neural tracing and nervous system-related drug delivery.

Sepsis is a life-threatening condition that occurs when the body is unable to effectively combat infection, leading to systemic inflammation and multi-organ failure. Interestingly, females exhibit lower sepsis incidence and improved clinical outcomes compared to males. However, the mechanisms underlying these sex-specific differences remain poorly understood. While sex hormones have been a primary focus, emerging evidence suggests that non-hormonal factors also play contributory roles. Despite sex differences in sepsis, clinical management is the same for both males and females, with treatment focused on combating infection using antibiotics and hemodynamic support through fluid therapy. However, even with these interventions, mortality remains high, highlighting the need for more effective and targeted therapeutic strategies. Sepsis-induced cardiomyopathy (SIC) is a key contributor to multi-organ failure and is characterized by left ventricular dilation and impaired cardiac contractility. In this review, we explore sex-specific differences in sepsis and SIC, with a particular focus on mitochondrial metabolism. Mitochondria generate the ATP required for cardiac function through fatty acid and glucose oxidation, and recent studies have revealed distinct metabolic profiles between males and females, which can further differ in the context of sepsis and SIC. Targeting these metabolic pathways could provide new avenues for sepsis treatment.

Glycocalyx is a hair-like structure covering the endothelium of the aqueous outflow pathway. While trabecular outflow is segmental circumferentially around the eye, regional differences in glycocalyx morphology remain largely unexplored. This study investigated glycocalyx variations in the different structures along the trabecular outflow pathway in high-flow (HF) and low-flow (LF) regions of bovine eyes.

Enucleated bovine eyes (n = 8) were perfused with fluorescein to identify HF and LF regions. The glycocalyx was labeled with Alcian Blue 8GX, and radial wedges from the anterior chamber angles of both HF and LF regions were processed for transmission electron microscopy. Glycocalyx thickness and coverage were quantified using ImageJ and compared between different outflow pathway locations in HF and LF regions. Glycocalyx measurements at intracellular (I-pores) and border pores (B-pores), the percentage of glycocalyx-unfilled pores, as well as the percentage of giant vacuoles (GVs) with and without I-pores with glycocalyx lining the inner membrane were evaluated.

Glycocalyx thickness and coverage did not differ significantly between HF and LF regions. However, thickness progressively increased from the proximal (trabecular meshwork) to the distal (episcleral veins) outflow pathway. In both I-pores and B-pores, the glycocalyx was present near the basal opening, edge, and center of the pores, with thickness increasing toward the center. No significant differences in the percentage of glycocalyx-filled pores were observed between HF and LF regions. However, the percentage of GVs with I-pores exhibiting glycocalyx lining the inner cellular membrane was significantly higher (100%) than that of those without I-pores (16%).

No regional differences were found between HF and LF regions, but glycocalyx thickness progressively increased from the proximal to the distal outflow pathway, potentially reflecting varying shear stress conditions. The significantly higher percentage of GVs with I-pores containing glycocalyx lining the inner cellular membrane compared to those without I-pores suggests a relationship between aqueous outflow dynamics and glycocalyx synthesis. These findings provide a morphological basis for future research on glycocalyx alterations in glaucoma and their impact on outflow resistance.

Coronavirus disease 2019 (COVID-19) has been associated with impaired endothelial and vascular function. We investigated whether intervention with glycocalyx dietary supplement (GDS), containing glucosamine sulfate and fucoidan, improves endothelial glycocalyx and vascular function after COVID-19 infection.

Fifty-seven convalescent patients 14 days after mild-to-moderate COVID-19 infection managed in an outpatient setting were randomized to receive GDS (n = 29) or placebo (n = 28) for 4 consecutive months. We measured at baseline and at 4 months: (a) perfused boundary region (PBR) of the sublingual microvessels with a diameter range of 4-25 μm, as a marker of endothelial glycocalyx integrity, (b) pulse wave velocity and augmentation index, (c) coronary flow reserve using Doppler echocardiography, and (d) malondialdehyde and protein carbonyls as oxidative stress markers.

Four months after treatment, patients who received GDS showed a greater reduction in PBR 4-25 μm (-6.8% vs. -1.3%), pulse wave velocity (-13.2% vs. -3%), augmentation index (-28.5% vs. -2.5%), malondialdehyde (-26% vs. -2.9%), protein carbonyls (-31.3% vs. -1%) and a greater increase in coronary flow reserve (12.9% vs. 1.6%) compared to placebo (p < .05). In the GDS group, the reduction in PBR 4-25 μm was associated with the corresponding decrease in pulse wave velocity (r = .31, p = .047), malondialdehyde, and protein carbonyls, as well as with the increase in coronary flow reserve (r = -.59, p = .008) at follow-up. Post-treatment, none of the patients under GDS reported post-COVID symptoms compared to 21.4% of the patients under placebo.

Four-month treatment with GDS may improve endothelial glycocalyx and vascular function after COVID-19 infection.

URL: https://www.

gov. Unique identifier: NCT05185934.

Endothelial cell (EC) glycocalyx (GCX) shedding from disturbed blood flow and chemical factors leads to low-density lipoprotein infiltration, reduced nitric oxide synthesis, vascular dysfunction and atherosclerosis. This study evaluates a therapy combining sphingosine-1-phosphate (S1P) and heparin (heparan sulfate derivative). We hypothesized that heparin/S1P co-treatment repairs mechanically damaged EC GCX in disturbed flow (DF) regions and restores anti-atherosclerotic mechanotransduction to treat cardiovascular disease.

We used a parallel-plate flow chamber to simulate flow conditions in vitro and a partial carotid ligation mouse model to mimic DF in vivo. Heparin and albumin-bound S1P were administered to assess their reparative effects on the endothelial GCX. Fluorescent staining, confocal microscopy, and ultrasound evaluated endothelial cell function and endothelial-dependent vascular function. Barrier functionality was assessed via macrophage uptake. Heparin/S1P mechanism-of-action insights were gained through fluid dynamics simulations and staining of GCX synthesis enzyme and S1P receptor. Statistical analyses validated the results.

The in vitro data showed that heparin/S1P therapy improves DF-conditioned ECs by restoring GCX and elevating vasodilator eNOS (endothelial-type nitric oxide synthase) expression. In vivo studies confirmed GCX degradation, vessel inflammation, hyperpermeability, and wall thickening in the mouse model's partially ligated left carotid artery. Heparin/S1P treatment restored GCX thickness and coverage, reduced inflammation and hyperpermeability, and inhibited vessel wall thickening.

This work introduces a new approach to regenerating the EC GCX and restoring its function in ECs under DF conditions, offering a groundbreaking solution for preventing cardiovascular diseases like atherosclerosis.

The glycocalyx of endothelial cells is a dynamic, gel-like layer of glycoproteins, proteoglycans, and glycolipids that lines the luminal surface of blood vessels, playing a critical role in vascular permeability, mechanotransduction, and protection against shear stress. In this study, we investigated the in vitro adhesion of giant unilamellar vesicles (GUVs) composed of 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) and 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC). Specifically, we examined mildly positively charged DOTAP-DMPC (20:80) GUVs, based on positively charged DOTAP and neutral DMPC but exhibiting an overall mild positive charge in physiological buffer, and neutral DMPC GUVs, which show a negative charge in physiological buffer. Adhesion to human umbilical vein endothelial cells (HUVEC) was studied under three culture conditions: dynamic (intact glycocalyx), static (underdeveloped glycocalyx), and glycocalyx-shed (degraded glycocalyx). Vesicles were produced via electroformation, stained with Texas Red dye, and perfused over endothelial cells at a controlled velocity to simulate slow blood flow. Adhesion was tracked using fluorescence microscopy combined with cell segmentation techniques. Adhesion of DOTAP-DMPC vesicles was significantly enhanced-by approximately 3.5-fold-on glycocalyx-shed cells compared to cells with an intact glycocalyx. In contrast, DMPC vesicles showed no adhesion under any condition. Analysis of vesicle size distributions revealed no significant differences between adherent and nonadherent vesicles or between DOTAP-DMPC and DMPC vesicles. These findings provide insights into the role of the endothelial glycocalyx in regulating adhesion, with potential implications for tumor cell interactions with the endothelium and mechanisms underlying DOTAP-based transfection.

Objectives: Children with nephrotic syndrome (NS) are prone to oral health issues due to immunosuppression and systemic inflammation, which may exacerbate their renal condition. The objective of this study was to evaluate the impact of a 12-year interdisciplinary collaboration between pediatric dentists and nephrologists on oral health in children with NS. Methods: A retrospective analysis of 80 NS patients-40 assessed in 2012 and 40 in 2024-was conducted using caries indices (dmft/DMFT), Plaque Index, and Gingival Index. Statistical tests assessed differences between groups (p < 0.05). Results: The prevalence of active caries significantly decreased (50% vs. 78%; p = 0.011), with fewer decayed permanent teeth (0.96 ± 1.56 vs. 2.66 ± 2.51; p = 0.003) and improved oral hygiene (good hygiene in 52.5% vs. 30%; p = 0.041) in the 2024 group. Gingivitis was less severe compared to 2012. Conclusions: Long-term interdisciplinary care significantly improved oral health in children with NS. These improvements may contribute to reduced systemic inflammation and better overall disease control. Integrating dental care into NS management is recommended to support long-term outcomes.

Chronic venous disease (CVD) is among the most common diseases in industrialized countries and has a significant socioeconomic impact. The diversity of clinical symptoms and manifestations of CVD pose major challenges in routine diagnosis and treatment. Despite the high prevalence and the huge number of venous surgical interventions performed every day, a substantial proportion of the etiopathogenesis remains unclear. There are several widely advocated and generally valid theories of "peri-capillary fibrin cuffs" and "white cell trapping hypothesis", which consider the role of venous reflux/obstruction, inflammation, vascular remodeling, hemodynamic changes, genetic and social risk factors. There are several specific provoking factors for the development of venous reflux: incompetence of the valve system, inflammation of the vascular wall, and venous hypertension. Over the past few years, increasing scientific data has demonstrated the link between oxidative stress, endothelial dysfunction, and vascular inflammation. High levels of oxidants and persistent inflammation can cause cumulative changes in hemodynamics, resulting in permanent and irreversible damage to the microcirculation and endothelial cells. Production of reactive oxygen species and expression of inflammatory cytokines and adhesion molecules are involved in a vicious cycle of venous wall remodeling. The interaction of ROS, and in particular, the superoxide anion radical, with nitric oxide leads to a decrease in NO bioavailability, followed by the initiation of prolonged vasoconstriction and hypoxia and impairment of vascular tone. This review addresses the role of ED, oxidative, and hemodynamic stress in the CVD mediation. Based on predefined inclusion and exclusion criteria, we conducted a systematic review of published scientific articles using PubMed, PMC Europe, Scopus, WoS, MEDLINE, and Google Scholar databases in the interval from 24 April 2002 to 1 April 2025. The current review included studies (n = 197) scientific articles, including new reviews, updates, and grey literature, which were evaluated according to eligibility criteria. The selection process was performed using a standardized form according to PRISMA rules, the manual search of the databases, and a double-check to ensure transparent and complete reporting of reviews. Studies had to report quantitative assessments of the relationship between vascular endothelial dysfunction, inflammation, oxidative stress, and shear stress in a chronic venous disease.

Acute renal dysfunction following hemorrhagic shock and resuscitation carries significant morbidity and mortality. While shock-induced shedding of the glycocalyx is well described within the pulmonary and splanchnic vasculature, less is known regarding early alterations to the glycocalyx of the renal microcirculation, particularly the glomerulus. We sought to evaluate the impact of hemorrhagic shock and resuscitation modalities on glomerular glycocalyx metabolism and function. We hypothesized that fresh frozen plasma resuscitation would attenuate glomerular glycocalyx shedding and reduce glomerular barrier dysfunction.

Male Sprague-Dawley rats were subjected to 60 minutes of hemorrhagic shock to 40% of baseline mean arterial pressure, followed by resuscitation with shed whole blood and either lactated Ringer's or fresh frozen plasma. Experimental groups included the following: (a) baseline, (b) post-hemorrhagic shock, (c) post-lactated Ringer's resuscitation, and (d) post-plasma resuscitation. Enzyme-linked immunosorbent assays and immunohistochemistry were used to evaluate alterations of syndecan-1 and hyaluronic acid within the glomerular glycocalyx. Urine protein concentration was measured as a surrogate for glomerular function, and expression of cubilin and megalin was quantified to evaluate renal tubule protein reabsorptive capacity.

Despite evidence of systemic glycocalyx shedding, hemorrhagic shock and resuscitation did not alter glomerular synedcan-1 expression. However, shock induced shedding of hyaluronic acid from the glomerular glycocalyx. While hyaluronic acid breakdown was exacerbated by crystalloid resuscitation, plasma utilization restored levels back to baseline. Urine protein concentration drastically increased following hemorrhagic shock and resuscitation with lactated Ringer's. By contrast, plasma administration reduced urine protein levels back to normal. Renal cortex cubilin and megalin expression did not differ among the experimental groups, suggesting that alterations in urine protein were driven by changes in glomerular function.

Plasma-based resuscitation appears to reverse shock-induced shedding of glomerular hyaluronic acid and attenuates glomerular barrier dysfunction. Differential shedding of the glomerular glycocalyx may represent a novel pathway in acute kidney injury pathophysiology.

The blood-brain barrier (BBB) glycocalyx is the dense layer of glycans and glycoconjugates that coats the luminal surface of the central nervous system (CNS) vasculature. Despite being the first point of contact between the blood and brain, not much is known about the BBB glycocalyx. Here, we performed a multi-omic investigation of the BBB glycocalyx which revealed a unique glycan landscape characterized by enrichment of sialic acid, chondroitin sulfate, and hyaluronan. We found that the BBB glycocalyx was thicker than glycocalyces in the peripheral vasculature and that hyaluronan was the major contributor to its ultrastructure. Using endothelial RNA sequencing, we found potential genetic determinants for these differences, including BBB enrichment of genes involved in sialic acid addition and peripheral enrichment of Tmem2 and Hyal2, the only known cell-surface hyaluronidases. Glycocalyx degradation and increases in vascular permeability are widely associated with inflammation. However, we found that the BBB glycocalyx remains largely unchanged in neuroinflammation during the experimental autoimmune encephalomyelitis (EAE) model of multiple sclerosis and that its degradation is not sufficient to alter BBB permeability in health. Moreover, we showed that CNS endothelial sialic acid removal delays onset of EAE, indicating that BBB glycocalyx sialic acid may contribute to the progression of neuroinflammation. These findings underscore the unique and robust nature of the BBB glycocalyx and provide targets and tools for future studies into its role in health and neuroinflammation.

Sepsis-induced acute kidney injury (S-AKI) is a common complication of sepsis. It occurs at high incidence and is associated with a high level of mortality in the intensive care unit (ICU). The pathophysiologic mechanisms underlying S-AKI are complex, and include renal vascular endothelial cell dysfunction. The endothelial glycocalyx (EG) is a polysaccharide/protein complex located on the cell membrane at the luminal surface of vascular endothelial cells that has anti-inflammatory, anti-thrombotic, and endothelial protective effects. Recent studies have shown that glycocalyx damage plays a causal role in S-AKI progression. In this review, we first describe the structure, location, and basic function of the EG. Second, we analyze the underlying mechanisms of EG degradation in sepsis and S-AKI. Finally, we provide a summary of the potential therapeutic strategies that target the EG.

Gliomas are the most common primary brain tumors in adults. However, glioblastoma is especially difficult to treat despite advancements in treatment. Therefore, new and more effective treatments are needed. The endothelial glycocalyx covers the luminal surface of the endothelium and plays an important role in vascular homeostasis. Tumor blood vessels normally have increased permeability, but some of them mimic normal cerebral blood vessels constituting the blood-brain barrier and retain drug-barrier function. Therefore, brain tumor vessels are considered to constitute the blood-tumor barrier. There are few reports on the endothelial glycocalyx in human brain tumor vessels. We aimed to visualize the endothelial glycocalyx in human brain tumor vessels and evaluate its microstructural differences in glioma vessels and normal capillaries. Surgical specimens from patients with glioma who underwent tumor resection at our institution were evaluated. We visualized the microstructures of the brain tumor vessels in human glioma specimens using electron microscopy with lanthanum nitrate. The endothelial glycocalyx was identified in the human glioma vasculature and its microstructure varied between the tumor margin and core. These variations may influence tumor angiogenesis and vascular remodeling, contributing to advancements in targeted therapies and diagnostics for human gliomas.

As the site of gas exchange, the lung is critical for organismal survival. It is also subject to continual environmental insults inflicted by pathogens, particles, and toxins. Sometimes, these insults result in structural damage and the initiation of an innate immune response. Operating in parallel, the immune response aims to eliminate the threat, while the repair process ensures continual physiological function of the lung. The inflammatory response and repair processes are thus inextricably linked in time and space but are often studied in isolation. Here, we review the interplay of innate immune cells and nonimmune cells during lung insult and repair. We highlight how cellular cross talk can fine-tune the circuitry of the immune response, how innate immune cells can facilitate or antagonize proper organ repair, and the prolonged changes to lung immunity and physiology that can result from acute immune responses and repair processes.

No longer viewed as a passive consequence of cellular activities, membrane curvature-the physical shape of the cell membrane-is now recognized as an active constituent of biological processes. Nanoscale topographies on extracellular matrices or substrate surfaces impart well-defined membrane curvatures on the plasma membrane. This review examines biological events occurring at the nano-bio interface, the physical interface between the cell membrane and surface nanotopography, which activates intracellular signaling by recruiting curvature-sensing proteins. We encompass a wide range of biological processes at the nano-bio interface, including cell adhesion, endocytosis, glycocalyx redistribution, regulation of mechanosensitive ion channels, cell migration, and differentiation. Despite the diversity of processes, we call attention to the critical role of membrane curvature in each process. We particularly highlight studies that elucidate molecular mechanisms involving curvature-sensing proteins with the hope of providing comprehensive insights into this rapidly advancing area of research.

The development of microcirculation imaging devices has significantly advanced our comprehension of the capillary environment's dynamics. Early research suggested that erythrocytes did not contact the vessel's inner surface due to the Fåhraeus effect, implying the presence of a covering on the endothelial cell surface. Subsequent electron microscopy studies revealed this layer to be a complex part of the vessel wall, now known as the endothelial glycocalyx (EG). The EG is a network of proteoglycans and glycoproteins bound to the endothelial membrane, incorporating soluble molecules from the endothelium and plasma. Over time, studies have elucidated the structure, function, and therapeutic targets of the glycocalyx, underscoring its pivotal role in vascular biology. The presence of cellular extensions of lung tissue cells in both vascular and nonvascular areas demonstrates the pivotal role of the glycocalyx in pulmonary vascular leak, surfactant dysfunction, impaired lung compliance and gas exchange abnormalities, which are hallmarks of acute respiratory distress syndrome (ARDS). It is of the utmost importance to elucidate the mechanisms underlying alveolocapillary glycocalyx degradation to develop efficacious treatments for ARDS, which has a mortality rate of 35 %. An understanding of the glycocalyx's role in vascular integrity provides a foundation for exploring new therapeutic avenues to mitigate lung injury and improve clinical outcomes in ARDS patients.

Glypican 4 (GPC4), a member of the cell surface heparan sulfate proteoglycan family, plays a crucial role in regulating various cell signaling and developmental processes. Its ability to be released from the cell surface into the bloodstream through shedding makes it a promising blood-based biomarker in health and disease. In this context, circulating GPC4 has been initially proposed as an insulin-sensitizing adipokine being linked with various conditions of insulin resistance. In addition, serum levels of GPC4 can indicate glycocalyx shedding and associated pathophysiological states, such as systemic inflammation. Particularly in a morbid and elderly population, increased GPC4 concentrations may reflect general organ dysfunction and an advanced state of multimorbidity, showing a strong association with the prognosis of severe conditions such as heart failure or advanced cancer. This comprehensive review is the first to summarize the existing scientific knowledge on the role of circulating GPC4 as a novel diagnostic and prognostic biomarker across different pathologic conditions. We also discuss in detail the putative underlying pathophysiological mechanisms behind these findings.

The glycocalyx and its associated endothelial surface layer which lines all cell membranes and most tissues, dwarfs the phospholipid membrane of cells in extent. Its major components are sulphated polymers like heparan and chondroitin sulphates and hyaluronic acid. These form a fuzzy layer of unknown structure and function. It has become increasingly clear that the ESL-GC complex must play many roles. We postulate it has a self-organised infrastructure that directs cell traffic, acts in defence against pathogens and other cells, and in diseases like diabetes, and heart disease, besides being a playground for a host of biochemical activity. Based on an analogous sulphated polymeric system Nafion, the fuel cell polymer, we suggest a model for the structure of the ESL-GC complex and how it functions. Taken together with parallel developments in physical chemistry, in nanobubbles, their stability in physiological media, and reactivity, we believe the model may throw light on a variety of phenomena, diabetes and some other diseases.

Glycosylation is the process by which glycans (i.e. 'sugars') are enzymatically attached to proteins or lipids to form glycoconjugates. Growing evidence points to glycosylation playing a central role in atherosclerosis. Glycosylation occurs in all human cells and post-translationally modifies many signalling molecules that regulate cardiovascular disease, affecting their binding and function. Glycoconjugates are present in abundance on the vascular endothelium and on circulating lipoproteins, both of which have well-established roles in atherosclerotic plaque development. Sialic acid is a major regulator of glycan function and therefore the process of sialylation, in which sialic acid is added to glycans, is likely to be entwined in any regulation of atherosclerosis. Glycans and sialylation regulators have the potential to present as new biomarkers that predict atherosclerotic disease or as targets for pharmacological intervention, as well as providing insights into novel cardiovascular mechanisms. Moreover, the asialoglycoprotein receptor 1 (ASGR1), a glycan receptor, is emerging as an exciting new regulator of lipid metabolism and coronary artery disease. This review summarises the latest advances in the growing body of evidence that supports an important role for glycosylation and sialylation in the regulation of atherosclerosis.

Nanoparticles are widely studied for delivering treatments to target tissues, but few have reached clinical use. Most nanoparticles encounter blood vessels on their way to target tissues. The inner surface of these vessels is lined with endothelial cells covered by a glycocalyx, an extracellular matrix rich in anionic glycans. The role of the glycocalyx in nanoparticle interactions is not well understood. Here, we demonstrate that endothelial cells need extended culture times to synthesize a mature glycocalyx. Our research shows that branched polyethyleneimine functionalized gold nanoparticles bind to endothelial cells expressing either a developing or mature glycocalyx, with the interaction involving hyaluronan and heparan sulfate. These nanoparticles are subsequently internalized. Similar results were seen with poly(L-arginine). A mature glycocalyx protects cells by reducing the toxicity of these cationic nanoparticles. In contrast, lipoic acid-functionalized gold nanoparticles are internalized by cells with a developing glycocalyx, but not a mature one. Poly(L-glutamic acid) only interacts with cells when major glycans in the glycocalyx are degraded. These findings highlight the complex relationship between nanoparticle charge and structure, and their effects on toxicity, binding, and uptake by endothelial cells. This offers important insights for improving nanoparticle interactions with blood vessels in health and disease. STATEMENT OF SIGNIFICANCE: Endothelial cells lining blood vessels form a barrier through which nanoparticles must cross to reach target tissues. These cells are covered with a layer called the glycocalyx, which is rich in anionic glycans. However, the role of the glycocalyx in how nanoparticles interact with cells remains underexplored. Our research revealed that cells with a mature glycocalyx internalize cationic nanoparticles and experience reduced cytotoxicity. Conversely, a mature glycocalyx prevents anionic nanoparticles from entering cells. These results suggest that the structure of both the nanoparticles and the glycocalyx should be considered in future studies to improve the use of nanoparticles for medical applications.

Endothelial cell dysfunction has a critical role in the pathophysiology of atherosclerosis. This study aims to uncover pivotal genes and pathways linked to endothelial cell dysfunction in atherosclerosis, as well as to ascertain the assumed causal effects and potential mechanisms.

Datasets relevant to endothelial cell dysfunction in atherosclerosis were collected and divided into training and validation sets. Following differential analysis, we constructed a protein-protein interaction (PPI) network and a molecular interaction map of common-differentially expressed genes (co-DEGs) with proteins known to be involved in atherosclerotic endothelial cell dysfunction. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genome (KEGG), and Gene Set Enrichment Analysis (GSEA) were also conducted. Moreover, human umbilical vein endothelial cells (HUVECs) were cultured in circumstances characterized by elevated glucose levels to establish a cellular injury model simulating atherosclerotic conditions, and quantitative Polymerase Chain Reaction (qPCR) experiments were conducted to validate the differences of co-DEGs. Subsequently, the Summary-data-based Mendelian Randomization (SMR) method was employed. Additionally, we employed the Western Blot (WB) technique to validate the differential expression of VAMP8. Finally, we identified the differential expression of VAMP8 in the validation set and further validated its differential expression by collecting fresh blood samples from 20 patients with atherosclerosis and 20 healthy individuals.

14 co-DEGs (FABP5, GULP1, COL4A5, VAMP8, FABP4, PFN2, ANGPT2, TFPI2, NUPR1, SULF1, FGF13, BASP1, EPB41L3, and PBK) were identified. SMR analysis confirmed 10 potential causal effect genes: PSRC1, VAMP8, FES, HNRNPUL1, CFDP1, SAP130, MDN1, OPRL1, UTP11, and HOXC4. The qPCR and WB experiments demonstrated that VAMP8 was significantly upregulated in the injured HUVECs group (p < 0.0001). Compared to the control group, VAMP8 was markedly increased in the blood samples of patients with atherosclerosis (p < 0.0001).

VAMP8 may potentially serve as a pathogenic gene in the process of endothelial dysfunction in atherosclerosis.

Intracellular delivery of proteins and small molecules is an important barrier in the development of strategies to deliver functional proteins and therapeutics into the cells to realize their full potential in biotechnology, biomedicine, cell-based therapies, and gene editing protein systems. Most of the intracellular protein delivery strategies involve the conjugation of cell penetrating peptides to enable the permeability of plasma membrane of mammalian cells to allow proteins to enter cytosol. The conjugations of small molecules such as (p-methylphenyl) glycine, pyrenebutyrate and cysteines are used for the same purpose. Molecular level interactions are governed mostly by ionic (cationic/anionic), covalent and noncovalent interactions with various molecular entities of glycocalyx matrix on plasma membrane lipid bilayer. Although the role of noncovalent interactions in cellular uptake is not fully understood, several recent advances have focused on the noncovalent interaction-based strategies of intracellular delivery of small molecules and proteins into mammalian cells. These are achieved by simple modification of protein surfaces with chemical moieties which can form noncovalent interactions other than hydrogen bonding. In this review, we describe the recent advances and the mechanistic aspects of intracellular delivery and role of noncovalent interactions in the cellular uptake of proteins and small molecules.

Flagellar-driven locomotion plays a critical role in the bacterial attachment and colonization of surfaces, contributing to the risks of contamination and infection. Extensive efforts to uncover the underlying principles governing bacterial motility near surfaces have relied on idealized assumptions about surrounding artificial surfaces. However, in the context of living systems, the role of cells from tissues and organs becomes increasingly critical, particularly in bacterial swimming and adhesion, yet it remains poorly understood. Here, we propose using biological surfaces composed of vascular endothelial cells to experimentally investigate bacterial motion and interaction behaviors. Our results reveal that bacterial trapping observed on inorganic surfaces is counteractively manifested with reduced radii of circular motion on cellular surfaces. Additionally, two distinct modes of bacterial adhesion were identified: tight and loose adhesion. Interestingly, the presence of living cells enhances bacterial surface enrichment, and imposed flow intensifies this accumulation via a bias-swimming effect. These results surprisingly indicate that physical effects remain the dominant factor regulating bacterial motility and accumulation at the single-cell-layer level in vitro, bridging the gap between simplified hydrodynamic mechanisms and complex biological surfaces with relevance to biofilm formation and bacterial contamination.

High dietary salt intake has powerful effects on cerebral blood vessels and has emerged as a risk factor for stroke and cognitive impairment. In mice, high salt diet (HSD) leads to reduced cerebral blood flow (CBF), tau hyperphosphorylation and cognitive dysfunction. However, it is still unclear whether the reduced CBF is responsible for the effects of HSD on tau and cognition. Capillary stalling has emerged as a cause of CBF reduction and cognitive impairment in models of Alzheimer's disease and diabetes. Therefore, we tested the hypothesis that capillary stalling also contributes to the CBF reduction and cognitive impairment in HSD. Using two-photon imaging, we found that HSD increased stalling of neutrophils in brain capillaries and decreased CBF. Neutrophil depletion reduced the number of stalled capillaries and restored resting CBF but did not prevent tau phosphorylation or cognitive impairment. These novel findings show that, capillary stalling contribute to CBF reduction in HSD, but not to tau phosphorylation and cognitive deficits. Therefore, the hypoperfusion caused by capillary stalling is not the main driver of the tau phosphorylation and cognitive impairment.

Understanding the intricate mechanisms underlying brain-related diseases hinges on unraveling the pivotal role of the blood-brain barrier (BBB), an essential dynamic interface crucial for maintaining brain equilibrium. This review offers a comprehensive analysis of BBB physiology, delving into its cellular and molecular components while exploring a wide range of in vivo and in vitro BBB models. Notably, recent advancements in 3D cell culture techniques are explicitly discussed, as they have significantly improved the fidelity of BBB modeling by enabling the replication of physiologically relevant environments under flow conditions. Special attention is given to the cellular aspects of in vitro BBB models, alongside discussions on advances in stem cell technologies, providing valuable insights into generating robust cellular systems for BBB modeling. The diverse array of cell types used in BBB modeling, depending on their sources, is meticulously examined in this comprehensive review, scrutinizing their respective derivation protocols and implications. By synthesizing diverse approaches, this review sheds light on the improvements of BBB models to capture physiological conditions, aiding in understanding BBB interactions in health and disease conditions to foster clinical developments.

The blood-brain barrier (BBB) is highly specialized to protect the brain from harmful circulating factors in the blood and maintain brain homeostasis1,2. The brain endothelial glycocalyx layer, a carbohydrate-rich meshwork composed primarily of proteoglycans, glycoproteins and glycolipids that coats the BBB lumen, is a key structural component of the BBB3,4. This layer forms the first interface between the blood and brain vasculature, yet little is known about its composition and roles in supporting BBB function in homeostatic and diseased states. Here we find that the brain endothelial glycocalyx is highly dysregulated during ageing and neurodegenerative disease. We identify significant perturbation in an underexplored class of densely O-glycosylated proteins known as mucin-domain glycoproteins. We demonstrate that ageing- and disease-associated aberrations in brain endothelial mucin-domain glycoproteins lead to dysregulated BBB function and, in severe cases, brain haemorrhaging in mice. Finally, we demonstrate that we can improve BBB function and reduce neuroinflammation and cognitive deficits in aged mice by restoring core 1 mucin-type O-glycans to the brain endothelium using adeno-associated viruses. Cumulatively, our findings provide a detailed compositional and structural mapping of the ageing brain endothelial glycocalyx layer and reveal important consequences of ageing- and disease-associated glycocalyx dysregulation on BBB integrity and brain health.

We examined how respiratory syncytial virus (RSV) particles circumvent the overlying glycocalyx on virus-infected A549 cells. The glycocalyx was detected using the lectin WGA-AL488 probe, and the antibodies anti-HS and anti-syndecan-4 that detect heparin sulphate (HS) and the syndecan-4 protein (SYND4) respectively. Imaging of RSV-infected cells provided evidence that the glycocalyx envelopes the virus filaments as they form, and that components of the glycocalyx such as HS moieties and SYND4 are displayed on the surface of the mature virus filaments. Recombinant expression of the G protein in these cells suggested that the G protein was trafficked into pre-existing filamentous cellular structures with a well-defined glycocalyx, further suggesting that the glycocalyx is maintained at the site of virus particle assembly. These data provide evidence that during RSV particle assembly the virus filaments become enveloped by the glycocalyx, and that the glycocalyx should be considered as a structural component of virus filaments.

The medical industry is an integral part of the delivery of healthcare. Collaboration between academic institutions, healthcare providers, and the industry are necessary but not devoid of flaws. This expert opinion article calls for closer attention to be paid by the medical industry to "what a frontline clinician needs" rather than relying solely on experts' opinions and stake holders' requests in planning future products and features. The need for the monitoring of tissue fluid accumulation is discussed from the point of view of practicing anaesthesiology and intensive care specialists in the context of the potential missed opportunity to have it be already available.

Preeclampsia (PE) is a gestational complication affecting 5% to 10% of all pregnancies. PE is characterized by hypertension and endothelial dysfunction, whose etiology involves, among other factors, alterations in the extracellular matrix (ECM) that can compromise vascular remodeling and trophoblast invasion, ie, processes essential for placental development. Endothelial dysfunction is caused by release of antiangiogenic factors, mainly a soluble fms-like tyrosine kinase-1 (sFlt-1), which antagonizes two endothelial angiogenic factors, the vascular endothelial growth factor (VEGF) and placental growth factor (PLGF). This angiogenic imbalance contributes to clinical symptoms including hypertension and multisystem dysfunction. This review aims to summarize recent advances in understanding PE, particularly with altered ECM components such as heparan sulfate proteoglycans, the glycosidase heparanase, fibronectin, collagen XVIII (endostatin), and metalloproteases. This comprehensive narrative review was conducted on PubMed from 1994 to 2024, focusing on articles on the pathophysiology of PE, particularly endothelial dysfunction caused by ECM modifications. The data shows a reduced expression of matrix metalloproteinases, increased collagen fragment XVIII, and significant changes in fibronectin associated with PE. Furthermore, endothelial dysfunction was associated with increased degradation of heparan sulfate chains from proteoglycans and increased sFlt-1. Understanding these ECM modifications is crucial for developing potential new therapeutic interventions that improve maternal and fetal outcome in PE.

The drivers of tissue necrosis in Mycobacterium ulcerans infection (Buruli ulcer disease) have historically been ascribed solely to the directly cytotoxic action of the diffusible exotoxin, mycolactone. However, its role in the clinically evident vascular component of disease aetiology remains poorly explained. We have now dissected mycolactone's effects on human primary vascular endothelial cells in vitro. We show that mycolactone-induced changes in endothelial morphology, adhesion, migration, and permeability are dependent on its action at the Sec61 translocon. Unbiased quantitative proteomics identified a profound effect on proteoglycans, driven by rapid loss of type II transmembrane proteins of the Golgi, including enzymes required for glycosaminoglycan (GAG) synthesis, combined with a reduction in the core proteins themselves. Loss of the glycocalyx is likely to be of particular mechanistic importance, since knockdown of galactosyltransferase II (beta-1,3-galactotransferase 6; B3GALT6), the GAG linker-building enzyme, phenocopied the permeability and phenotypic changes induced by mycolactone. Additionally, mycolactone depleted many secreted basement membrane components and microvascular basement membranes were disrupted in vivo during M. ulcerans infection in the mouse model. Remarkably, exogenous addition of laminin-511 reduced endothelial cell rounding, restored cell attachment and reversed the defective migration caused by mycolactone. Hence supplementing mycolactone-depleted extracellular matrix may be a future therapeutic avenue, to improve wound healing rates.

Ischemic stroke, a blockage in the vasculature of the brain that results in insufficient blood flow, is one of the world's leading causes of disability. The cascade of inflammation and cell death that occurs immediately following stroke drives vascular and functional loss that does not fully recover over time, and no FDA-approved therapies exist that stimulate regeneration post-stroke. We have previously developed a hydrogel scaffold that delivered heparin nanoparticles with and without VEGF bound to their surface to promote angiogenesis and reduce inflammation, respectively. However, the inclusion of the naked heparin nanoparticles warranted concern over the development of bleeding complications. Here, we explore how microporous annealed particle (MAP) scaffolds functionalized with VEGF coated heparin nanoparticles can both reduce inflammation and promote angiogenesis - without the inclusion of free heparin nanoparticles. We show that our updated design not only successfully promotes de novo tissue formation, including the development of mature vessels and neurite sprouting, but it also leads to functional improvement in a photothrombotic stroke model. In addition, we find increased astrocyte infiltration into the infarct site correlated with mature vessel formation. This work demonstrates how our biomaterial design can enhance endogenous regeneration post-stroke while eliminating the need for excess heparin.

The LINC (linker of nucleoskeleton and cytoskeleton) complex is a critical component of the cellular architecture that bridges the nucleoskeleton and cytoskeleton and mediates mechanotransduction to and from the nucleus. Though it plays important roles in all blood vessels, it is in arterioles that this complex plays a pivotal role in maintaining endothelial cell integrity, regulating vascular tone, forming new microvessels and modulating responses to mechanical and biochemical stimuli. It is also important in vascular smooth muscle cells and fibroblasts, where it possibly plays a role in the contractile to secretory phenotypic transformation during atherosclerosis and vascular ageing, and in fibroblasts' migration and inflammatory responses in the adventitia. Physiologically, the LINC complex contributes to the stability of arteriolar structure, adaptations to changes in blood flow and injury repair mechanisms. Pathologically, dysregulation or mutations in LINC complex components can lead to compromised endothelial function, vascular remodelling and exacerbation of cardiovascular diseases such as atherosclerosis (arteriolosclerosis). This review summarizes our current understanding of the roles of the LINC complex in cells from arterioles, highlighting its most important physiological functions, exploring its implications for vascular pathology and emphasizing some of its functional characteristics in endothelial cells. By elucidating the LINC complex's role in health and disease, we aim to provide insights that could improve future therapeutic strategies targeting LINC complex-related vascular disorders.

Returning to the moon and traveling to Mars represent the main targets of human space exploration missions within the upcoming decades. Comparable to microgravity, partial gravity in these destinations is assumed to dysregulate immune functions, thereby threatening astronauts´ health. To investigate the impact of partial gravity on immune cell attachment to vessel endothelia, THP-1 cells and HUVEC cell layers were monitored in a flow chamber system during parabolic flight in lunar (0.16 g) or Martian (0.38 g) gravity. Focus was set on floating speed, cell adhesion, surface molecule expression and cytoskeletal reorganization under basal and TNF-induced inflammatory environment. Floating speed of THP-1 cells was increased in partial gravity, which was accompanied by a successively lower adhesion to the endothelial HUVEC cells. Expression levels of the adhesion markers Mac-1 on THP-1 cells as well as ICAM-1 on HUVECs were found elevated in lunar and Martian gravity, which was aggravated by TNF. Analysis of cytoskeletal organization in HUVECs revealed reduced intracellular F-actin microfilament networks and a stronger cell directionality with stress fiber alignment at cell borders in partial gravity, which was intensified by TNF. In summary, altered immune cell - endothelium interactions as quantified in partial gravity conditions show similarities to cellular behavior in microgravity. However, the different magnitudes of effects in dependence of gravitational level still need to be assessed in further investigations.