Endothelial Cell Morphology Research Articles

Endothelial dysfunction in Marfan syndrome mice is restored by resveratrol

Patients with Marfan syndrome (MFS) develop thoracic aortic aneurysms as the aorta presents excessive elastin breaks, fibrosis, and vascular smooth muscle cell (vSMC) death due to mutations in the FBN1 gene. Despite elaborate vSMC to aortic endothelial cell (EC) signaling, the contribution of ECs to the development of aortic pathology remains largely unresolved. The aim of this study is to investigate the EC properties in Fbn1C1041G/+ MFS mice. Using en face immunofluorescence confocal microscopy, we showed that EC alignment with blood flow was reduced, EC roundness was increased, individual EC surface area was larger, and EC junctional linearity was decreased in aortae of Fbn1C1041G/+ MFS mice. This modified EC phenotype was most prominent in the ascending aorta and occurred before aortic dilatation. To reverse EC morphology, we performed treatment with resveratrol. This restored EC blood flow alignment, junctional linearity, phospho-eNOS expression, and improved the structural integrity of the internal elastic lamina of Fbn1C1041G/+ mice. In conclusion, these experiments identify the involvement of ECs and underlying internal elastic lamina in MFS aortic pathology, which could act as potential target for future MFS pharmacotherapies.

Carboxy-terminal telopeptide levels of type I collagen hydrogels modulated the encapsulated cell fate for regenerative medicine

The cellular microenvironment has a profound impact on cell proliferation, interaction, and differentiation. In cell encapsulation for disease therapy, type I collagen is an important biomaterial due to its ability to mimic the extracellular matrix. Telopeptides (carboxy-terminal, CTX, and amino-terminal, NTX) protruding from the triple helix structure of type I collagen are cross-link sites, but also mediate the signal transmission in tissue homeostasis. It is worth investigating the features of the hydrogel microenvironment shaped by the tissue-derived type I collagen with various telopeptide levels, which is paramount for encapsulated cell development. Here, we found the fate of encapsulated human adipose-derived stem cells (hADSCs) and human umbilical vein endothelial cells (HUVECs) behaved differently towards decreasing CTX levels in the collagen hydrogels. Even among collagen hydrogels with a small magnitude of CTX variation, similar stiffness and microstructure, the apparent CTX modulation on the proliferation, cell-interaction, and genes expression of encapsulated hADSCs, as well as morphology and tubule structure formation of endothelial cells were observed, suggesting the biological roles of CTX and its modulation on microenvironment for cell development.

Corneal endothelial cell morphology and optical coherence tomography findings in children with type 1 diabetes mellitus.

To evaluate central macular thickness, retinal nerve fibre layer thickness, corneal endothelial cell density and central corneal thickness in children with Type 1 Diabetes Mellitus (DM). Thirty children with Type 1 DM and 30 age-matched children as controls were examined. Central macular thickness (CMT) and four quadrants of retinal nerve fibre layer thickness (RNFLT) were measured by optical coherence tomography (OCT), while endothelial cell density (ECD), coefficient of variation in cell size (CV), hexagonality (HEX) and central corneal thickness (CCT) were measured by noncontact specular microscopy. The mean ECD was 2810.77 ± 273.47 cells/mm2 and the mean hexagonality ratio was 49.77 ± 13.2 both were significantly lower (p < 0.001, p = 0.037 respectively) in the diabetic group compared to the control group. The mean CV was 37.6 ± 7.27, it was significantly higher (p = 0.024) in the diabetic group than the control group. The mean corneal thickness was greater in the DM group than the controls, but the difference was not significant (p = 0.176). There were no significant differences in CMT or RNFLT between the two groups. There was a negative correlation between the HbA1c levels and the thickness of the superior quadrants of RNFL (r = -0.406, p = 0.026). The duration of diabetes had no significant correlations with the corneal and retinal parameters. The diabetic children had changes in corneal endothelial morphology and there was a negative correlation between HbA1c levels and superior quadrant of RNFLT.

Effects of Intracameral Drugs and Dyes on Corneal Endothelial Cell Apoptosis in a Rat Model: An In Vivo and In Vitro Analysis.

To evaluate the effects of intracameral drugs and dyes on rat corneal endothelial apoptosis and cell morphology. The right eyes of 72 rats were injected intracamerally with 1% lidocaine, 0.01% adrenaline, triamcinolone acetonide (TA) 4 mg/mL, 1% trypan blue (TB), 0.5% indocyanine green (ICG), and fortified balanced salt solution as control. Corneal samples were taken 1 day and 1 week post-injection. Corneal endothelial apoptosis was assessed by the TUNEL technique, and the ratio of apoptotic cells in each group was compared with the control. Corneal endothelial cell morphology was evaluated in each specimen by transmission electron microscopy. The mean apoptotic endothelial cell ratio was significantly higher at 1 day and 1 week after intracameral adrenaline injection when compared to controls (p=0.03 and 0.021, respectively). TB caused a significantly higher apoptotic cell ratio when compared to controls at 1 week after injection (p=0.043). Lidocaine caused a higher apoptotic cell ratio compared to TA and ICG at 1 week, although not statistically significant (p=0.058, 0.09, 0.69, respectively). In all experimental specimens, transmission electron microscopy showed morphological changes associated with apoptosis. This study showed that intracameral adrenaline, TB, and lidocaine injections may have toxic effects on corneal tissue, as indicated by ultrastructural and histopathological alterations. Therefore, these agents should be used with caution in intraocular surgery.

Effectiveness of selective laser trabeculoplasty and its safety on corneal endothelium in patients with primary open angle glaucoma and ocular hypertension.

The aim of this study was to evaluate the effectiveness of selective laser trabeculoplasty (SLT) in patients with primary open-angle glaucoma (POAG) and ocular hypertension (OHT), and to assess its effects and safety on corneal endothelial cell morphology. Forty patients with POAG (15 cases, 23 eyes) and OHT (25 cases, 48 eyes) were prospectively evaluated. All cases underwent pachymetry, goniolens examination, Humphrey Visual Field Test, Optical Coherence Tomography, and Corneal Confocal Microscopy measurements. Patients whose intraocular pressure (IOP) was more than 21 mmHg in POAG and OHT, underwent SLT. SLT treatment was applied to the inferior 180° region of the trabecular meshwork. Central corneal thickness (CCT), IOP, and specular microscopy examinations were taken at the visits before and after the first day, first week, first month, third and sixth months of the SLT procedure. Retinal nerve fiber layer (RNFL) thickness measurements and visual field tests were performed before and six month post-SLT. Results of the pre-SLT and post-SLT measurements were compared and a P value of lower than 0.05 was considered statistically significant. The results of IOP measurements at first week, first month, third, and sixth month post-SLT were significantly lower than the results before and post-SLT first day. No significant difference was found between the results of initial and final measurements of CCT and specular microscopic corneal endothelial cell morphology evaluation. There was no statistically significant difference between the results of initial and final measurements for RNFL thickness and the results of the visual field tests. SLT is an effective treatment modality for reducing IOP in patients with POAG and OHT and is also a safe procedure in terms of the entirety of corneal endothelial cell morphology.

Non-Invasive Pulsatile Shear Stress Modifies Endothelial Activation; A Narrative Review.

The monolayer of cells that line both the heart and the entire vasculature is the endothelial cell (EC). These cells respond to external and internal signals, producing a wide array of primary or secondary messengers involved in coagulation, vascular tone, inflammation, and cell-to-cell signaling. Endothelial cell activation is the process by which EC changes from a quiescent cell phenotype, which maintains cellular integrity, antithrombotic, and anti-inflammatory properties, to a phenotype that is prothrombotic, pro-inflammatory, and permeable, in addition to repair and leukocyte trafficking at the site of injury or infection. Pathological activation of EC leads to increased vascular permeability, thrombosis, and an uncontrolled inflammatory response that leads to endothelial dysfunction. This pathological activation can be observed during ischemia reperfusion injury (IRI) and sepsis. Shear stress (SS) and pulsatile shear stress (PSS) are produced by mechanical frictional forces of blood flow and contraction of the heart, respectively, and are well-known mechanical signals that affect EC function, morphology, and gene expression. PSS promotes EC homeostasis and cardiovascular health. The archetype of inducing PSS is exercise (i.e., jogging, which introduces pulsations to the body as a function of the foot striking the pavement), or mechanical devices which induce external pulsations to the body (Enhanced External Pulsation (EECP), Whole-body vibration (WBV), and Whole-body periodic acceleration (WBPA aka pGz)). The purpose of this narrative review is to focus on the aforementioned noninvasive methods to increase PSS, review how each of these modify specific diseases that have been shown to induce endothelial activation and microcirculatory dysfunction (Ischemia reperfusion injury-myocardial infarction and cardiac arrest and resuscitation), sepsis, and lipopolysaccharide-induced sepsis syndrome (LPS)), and review current evidence and insight into how each may modify endothelial activation and how these may be beneficial in the acute and chronic setting of endothelial activation and microvascular dysfunction.

Effect Assessment of Aurantio-Obtusin on Novel Human Renal Glomerular Endothelial Cells Model Using a Microfluidic Chip.

Cassiae semen is widely used as a raw material of health food. Anthraquinone compounds, the main components in cassiae semen, have been reported to show nephrotoxicity. Aurantio-obtusin (AO) is a major anthraquinone compound extracted from cassiae semen. This study investigates the effects of AO on the morphology and physiological function of human renal glomerular endothelial cells (HRGECs) on a microfluidic chip device for the first time. HRGECs were cultured on a microfluidic plate and exposed to a series of AO concentrations. Compared with traditional 96-well culture, HRGECs cultured on the microfluidic chip appeared to better mimic the glomerular microenvironment in vivo. AO induced different degrees of damage to cellular morphology and physiological function. The leakage of lactate dehydrogenase (LDH), as well as the secretion of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), transforming growth factor-β1 (TGF-β1), and monocyte chemoattractant protein 1 (MCP-1), increased in the AO treated groups. At the same time, cell viability and expression of ZO-1 in the AO treated groups decreased in a dose-dependent manner. The innovative device enables direct visualization and quantification to evaluate the cytotoxic effects of AO on HRGECs, and provides a useful visual in vitro model for studying health effect of health food.

Comparison of Corneal Endothelium in Patients with Uveitis and Healthy Subjects Using Specular Microscopy

Objective: To evaluate changes in corneal endothelium in patients with anterior uveitis by using specular microscopy and compare it with the corneal endothelium of healthy subjects.&#x0D; Study Design: Comparative cross-sectional study.&#x0D; Place and Duration of Study: Armed Forces Institute of Ophthalmology, Rawalpindi Pakistan, from Jun 2018 to Jun 2019.&#x0D; Methodology: This study included 65 eyes of 65 patients with anterior uveitis and 65 eyes of healthy subjects. Corneal endothelium was assessed in all participants of both Groups by using Specular microscopy. Corneal endothelial markers like central corneal thickness (CCT), cell density (CD), cell minimum area, cell maximum area, the average of cell size, percent of hexagonality (HEX%), coefficient of variation (CV) were compared in both Groups. The Uveitis Group was further subdivided into active and inactive Uveitis Subgroups, and corneal endothelium markers were also compared in these Subgroups.&#x0D; Results: The mean endothelial cell density was 2626.0±413.0 cells/mm² in patients with uveitis, whereas cell density was 2766.0±327.0 cells/mm² in healthy subjects. The difference in endothelial cell density was statistically significant between the Groups (p=0.03). The difference between central corneal thickness, maximum cell area, average cell size and hexagonality values between Groups was also statistically significant, whereas there was no difference in terms of cell minimum area between these Groups. There was no statistically significant difference between inactive and active uveitis when corneal endothelial markers were compared between them (p&gt;0.05).&#x0D; Conclusion: There was an effect of uveitis on central corneal thickness, endothelial cell density, cell size, and morphology compared to healthy subjects,......

Impact of spatial and temporal stability of flow vortices on vascular endothelial cells.

Intracranial aneurysms (IAs) are pathological dilations of cerebrovascular vessels due to degeneration of the mechanical strength of the arterial wall, precluded by altered cellular functionality. The presence of swirling hemodynamic flow (vortices) is known to alter vascular endothelial cell (EC) morphology and protein expression indicative of IAs. Unfortunately, less is known if vortices with varied spatial and temporal stability lead to differing levels of EC change. The aim of this work is to investigate vortices of varying spatial and temporal stability impact on ECs. Vortex and EC interplay was investigated by a novel combination of parallel plate flow chamber (PPFC) design and computational analysis. ECs were exposed to laminar (7.5 dynes/[Formula: see text] wall shear stress) or low (<1 dynes/[Formula: see text]) stress vortical flow using PPFCs. Immunofluorescent imaging analyzed EC morphology, while ELISA tests quantified VE-cadherin (cell-cell adhesion), VCAM-1 (macrophage-EC adhesion), and cleaved caspase-3 (apoptotic signal) expression. PPFC flow was simulated, and vortex stability was calculated via the temporally averaged degree of (volume) overlap (TA-DVO) of vortices within a given area. EC morphological changes were independent of vortex stability. Increased stability promoted VE-cadherin degradation (correlation coefficient r = [Formula: see text]0.84) and 5-fold increased cleaved caspase-3 post 24 h in stable (TA-DVO 0.736±0.05) vs unstable (TA-DVO 0.606[Formula: see text]0.2) vortices. ECs in stable vortices displayed a 4.5-fold VCAM-1 increase than unstable counterparts after 12 h. This work demonstrates highly stable disturbed flow imparts increased inflammatory signaling, degraded cell-cell adhesion, and increased cellular apoptosis than unstable vortices. Such knowledge offers novel insight toward understanding IA development and rupture.

Comparison of corneal, endothelial, and anterior segment parameters in eyes with and without pigment dispersion

BackgroundTo evaluate the differences of central corneal thickness (CCT), corneal endothelial cell morphology, and biometric parameters of the anterior chamber and iridocorneal angle in pigment dispersion syndrome (PDS), pigmentary glaucoma (PG), primary open-angle glaucoma (POAG), and healthy group. MethodsTwenty- three eyes in the PDS group, 39 eyes in the PG group, 33 eyes in the POAG group, and 45 eyes in the healthy control group were included in this cross-sectional retrospective study. Corneal endothelial cell density, the coefficient of variation in the cell area, hexagonality, and central corneal thickness (CCT) were obtained by specular microscopy (CEM 530, NIDEK, Japan). Anterior chamber depth (ACD), iridocorneal angle parameters and CCT were measured by anterior segment optical coherence tomography (AS-OCT) (Visante OCT, Carl Zeiss AG, Germany). ResultsThe mean CCT via specular microscopy (SM-CCT) was measured at the thinnest value in the PG group (531.20 ± 34.91 µm) and the thickest in the control group (569.13 ± 37.52 µm). CV value was higher in PG (34.65 ± 6.84) and POAG group (34.27 ± 9.93) and lower in control group (28.82 ± 5.18) (p = 0,005). The mean AS-OCT-CCT was the thinnest in the PG group (513.61 ± 39.94 µm), and the thickest in the control group (547.04 ± 36.72 µm) (p = 0.001). All parameters of the iridocorneal angle were larger in the PDS and PG groups (p < 0.001). In the correlation analysis of the glaucomatous eyes, a negative correlation between the C/D ratio and SM-CCT and AS-OCT-CCT (p = 0.037, p = 0.017, respectively) and a positive correlation between the pRNFL thickness and AS-OCT- CCT (p = 0.002). ConclusionCCT values obtained with both SM and AS-OCT were found to be significantly thinner in PG cases. CV, which is the measure of polymegatism, was found to be significantly higher in PG cases.

Effect of mechanical stretching and substrate stiffness on the morphology, cytoskeleton and nuclear shape of corneal endothelial cells

Due to the limited capacity of corneal endothelial cells (CECs) division, corneal endothelial diseases have become a great challenge. The cornea is subjected to various mechanical stimuli in vivo, which may have a positive or negative influence. Thus, it is significant to gain an insight into the mechanism of mechanobiology of CECs for seeking more possible treatment. The purpose of this study was to determine the impacts of mechanical stretch and substrate stiffness on the morphology and fundamental cell behavior of CECs. Rabbit corneal endothelial cells (RCECs) were subjected to a 5% mechanical stretch or cultured on substrates of different stiffness. The impacts of mechanical stimulus on cell area, aspect ratio, circularity, cell density, nuclear shape, cytoskeleton, and cell viability were investigated. The expressions of the corneal endothelium-related markers ZO-1 and Na+/K+-ATPase were also evaluated by confocal immunofluorescence microscopy in the stiffness group. Our results suggested that mechanical stretch promoted the rearrangement of the cytoskeleton while decreasing the cell circularity, nuclear area, and cell density as well as cell viability. RCECs cultured on 10 ​kPa substrates, which was close to the physiological stiffness of rabbit Descemet's membrane (DM), showed better cell morphology, more stable actin cytoskeleton assembly, and more robust expression of the functional marker compared with other softer or stiffer substrates. In summary, mechanical stretch and substrate stiffness have profound influences on the morphology and function of CECs, which may have implications for the understanding and possible treatment of corneal endothelial diseases.

COMPARISON OF CHANGES IN ENDOTHELIAL CELL DENSITY AND MORPHOLOGY AFTER MANUAL SMALL INCISION CATARACT SURGERY IN DIABETICS VS. NON-DIABETICS – A PROSPECTIVE STUDY

Purpose: To compare the changes in endothelial cell density (ECD) and morphology after manual small incision cataract surgery (MSICS) in diabetics vs. non-diabetics All consecutive consenting adults Methods: with age related cataract &lt; grade 2 nuclear sclerosis undergoing MSICS were recruited. Specular microscopy was done to document the ECD, coefcient of variation (CoV) and percentage of hexagonality in all eyes before and 1 month after MSICS. The primary outcome measure was the difference in ECD between diabetics and non-diabetics at 1 month compared to baseline. One hundred eyes of 100 Results: patients were recruited in the diabetic (n=50) and non-diabetic (n=50) groups and completed 1 month follow up. There were no differences at baseline, though non-diabetics had marginally higher counts (p=0.07). The mean ECD decreased signicantly in both groups (p&lt;0.001 for both) but this drop was signicantly more in diabetics (566 + 137cells/mm3, 95% CI= 527 to 604) compared to non-diabetics (347 + 94cells/mm3, 95% CI= 320 to 373) (p&lt;0.001). The percentage hexagonality decreased slightly after MSICS in both, although by the same magnitude (-6.4 + 3.7 in diabetics vs. -5.2 + 3.9%, p=0.09) where as the CoV changed more (-5.8 + 5.5 in non-diabetics vs. 2.9 + 2.6, p&lt;0.001) in the diabetic group. Eyes Conclusions: with softer cataracts in diabetics undergoing MSICS experience much greater endothelial cell loss at 1 month, though changes in the morphology such as percentage hexagonality and CoV are not as marked.

Structural abnormalities in the non-dilated ascending aortic wall of bicuspid aortic valve patients

BackgroundA bicuspid aortic valve (BAV) is the most common congenital cardiac malformation. The development of the aortic valve is closely related to the development of the ascending aorta, associated with structural differences in the bicuspid aorta. Here we describe the non-dilated ascending aortic wall in bicuspid aortic valve patients. MethodsBAV (n=41) and tricuspid aortic valve (TAV) (n=18) non-dilated ascending aortic wall samples were studied. We investigated the following features of the aortic wall: vessel wall thickness, endothelial cell morphology, atherosclerosis, and elastic lamellae organization. Medial pathologic features encompassing elastic fiber thinning, fragmentation and degeneration, overall medial degeneration, mucoid extracellular matrix accumulation, and smooth muscle cell nuclei loss were studied. Furthermore, we included apoptosis, periaortic inflammation, and the level of expression of differentiated vascular smooth muscle cells. ResultsThe non-dilated BAV ascending aortic wall is characterized by a significantly thinner intimal layer, without features of atherosclerosis (P<.001). The medial layer is significantly thicker (P<.001) with more mucoid extracellular matrix accumulation (P<.001). All other medial pathologic features were more prominent in the TAV (P<.001). The media has significantly less differentiated vascular smooth muscle cells (P<.001) between the neatly regulated elastic lamellae which are thinner in the BAV as compared to the TAV (P<.0001). ConclusionsThe BAV ascending aorta without dilatation is characterized by a differentiation defect of vascular smooth muscle cells in the media and a significantly thinner intimal layer without overt pathologic features.

Corneal endothelial morphology of healthy myopic Malaysian children of Chinese ethnicity aged 8-9 years and its association with axial length

Background: This is a cross-sectional study to further understand the effects of axial length elongation on the corneal endothelial cell's morphology in myopic children. Abnormal changes in the morphology of corneal endothelium are indicators of corneal stress or instability which could be linked to myopia. Methods: 111 school children comprising of 37 emmetropes, 37 mild myopes and 37 moderate myopes aged 8-9 years old were recruited. Visual acuity was measured using the LogMar chart, cycloplegic refraction was determined using an open-field autorefractor (Grand Seiko WAM-5100, Hiroshima, Japan) and refined using subjective refraction. Morphology of corneal endothelial cells [endothelial cell density, coefficient of variation, hexagonality and central corneal thickness] was evaluated using a non-contact specular microscope (Topcon SP-2000P). Axial length was measured with A-scan ultrasound biometry (PacScan Plus, Sonomed Escalon, NY). The correlation between morphology of corneal endothelial cells and axial length were assessed using Pearson Correlation and Linear regression analysis. Results: There was no significant difference in corneal endothelial cells and axial length between gender (p>0.05). Significant reduction in endothelial cells density and hexagonality and increased coefficient of variation was found in eyes of higher myopic power which had longer axial when compared to emmetropes (p<0.001). Except for central corneal thickness, all corneal endothelial cells parameters correlated significantly with axial length (p<0.05). For every 1mm increase in axial length, endothelial cells density decreased by 73.27cells/mm 2, hexagonality decreased by 2.32% and coefficient of variation increased by 1.75%. Conclusions: There were significant changes in morphology of cornea endothelial cells in young moderate myopic children of Chinese ethnicity at 8-9 years of age. This result provides normative data for Malaysian children of Chinese ethnicity that can be used for comparison and reference for clinical procedures, thereby facilitating decision-making with respect to interventions for myopia control, especially in prescribing contact lens for children.

Microengineering 3D Collagen Hydrogels with Long-Range Fiber Alignment.

Aligned collagen I (COL1) fibers guide tumor cell motility, influence endothelial cell morphology, control stem cell differentiation, and are a hallmark of cardiac and musculoskeletal tissues. To study cell response to aligned microenvironments in vitro, several protocols have been developed to generate COL1 matrices with defined fiber alignment, including magnetic, mechanical, cell-based, and microfluidic methods. Of these, microfluidic approaches offer advanced capabilities such as accurate control over fluid flows and the cellular microenvironment. However, the microfluidic approaches to generate aligned COL1 matrices for advanced in vitro culture platforms have been limited to thin "mats" (<40 µm in thickness) of COL1 fibers that extend over distances less than 500 µm and are not conducive to 3D cell culture applications. Here, we present a protocol to fabricate 3D COL1 matrices (130-250 µm in thickness) with millimeter-scale regions of defined fiber alignment in a microfluidic device. This platform provides advanced cell culture capabilities to model structured tissue microenvironments by providing direct access to the micro-engineered matrix for cell culture.

Effects of exogenous hydrogen sulfide on pulmonary hypertension in rabbits with endotoxic shock

Objective: To investigate the effects of exogenous hydrogen sulfide (H2S) on pulmonary vascular reactivity induced by endotoxic shock (ES) in rabbits. Methods: In this experiment, the model of endotoxic shock (ES) was induced by injection of lipopolysaccharides (LPS) to New Zealand big eared white rabbit through jugular vein (8 mg/0.8 ml/kg), the intervention was performed by H2S donor(sodium hydrosulfide, NaHS) which was injected intraperitoneally (28 μmol/kg) 15 min in advance. New Zealand rabbits were randomly divided into 4 groups(n=8):control group, LPS group, LPS+NaHS group and NaHS group. The changes of mean arterial pressure (MAP) and mean pulmonary arterial pressure (MPAP) were detected. The tension of pulmonary artery ring (PARs) was detected byin vitro vascular ring technique. The ultrastructure of pulmonary artery wall and pulmonary artery endothelial cells were observed by light microscope and scanning electron microscope. Results: ①MAP was decreased while MPAP was increased in rabbits after LPS injection, and ES animal model was established successfully. Compared with LPS group, mPAP of rabbit in LPS+NaHS group was decreased significantly (all P＜0.05). ②Compared with normal control group, pulmonary artery of rabbits in LPS group had an increased contractile response to phenylephrine (PE) and a decreased relaxation response to acetylcholine (ACh) (both P＜0.01); Compared with LPS group, pulmonary artery of rabbits in LPS+NaHS group had a decreased contractile response to PE and an increased relaxation response to ACh (both P＜0.05). ③Under light microscope, the structure of vascular endothelial cells was continuous in the normal control group, the elastic fibers were intact in the subcutaneous layer, and the smooth muscle layer was arranged neatly. LPS can shed some of the pulmonary artery endothelial cells, break the subcutaneous elastic fibers, and disorder the smooth muscle layer structure. Compared with LPS group, the injury of pulmonary artery wall in LPS+NaHS group was ameliorated. The morphology of pulmonary artery wall was normal in NaHS group. It is showed that some endothelial cells of pulmonary artery were missing in LPS group by Scanning electron microscopy. The morphology of pulmonary artery endothelial cells in LPS+NaHS group was similar to that in the control group: slightly widened intercellular space was observed, and no cell exfoliation was observed. Conclusion: These results suggest that exogenous H2S can protect pulmonary artery endothelial cells and regulate the reactivity changes of pulmonary artery during ES, which may be one of the mechanisms reducing PAH in ES rabbits.

Case-control study of corneal topography and specular microscopy parameters in JIA patients with and without ocular involvement.

To assess corneal topography and specular microscopy values in juvenile idiopathic arthritis-related uveitis (JIA-U). This case-control study included 30 eyes from JIA-U patients, 20 eyes from JIA patients, and 50 eyes from age- and gender-matched healthy subjects. Patients with a history of ocular diseases or intraocular surgery were excluded. Corneal topography maps (Pentacam HR) and specular microscopy images (CellChek SL) were obtained. The measurements of the groups were compared. Keratometric astigmatism was higher in the JIA-U group than in the control group (p = 0.040). Patients with astigmatism greater than 1.50D were more common in the JIA-U group than in the control group (p = 0.026). The JIA-U group had higher anterior and posterior elevation values than the control group (p = 0.006, p = 0.025). The density of endothelial cells, coefficient of variation, and hexagonality did not change across groups (p = 0.465, p = 0.096, p = 0.869). The total number of exacerbations and the duration of anterior chamber inflammation were both positively correlated with posterior elevation (r = 0.600, p 0.001; r = 0.583, p 0.001). The age of diagnosis was found to be negatively correlated with anterior elevation (r = -0.412, p = 0.021). Corneal astigmatism, as well as anterior and posterior elevation values, were all higher in JIA-U patients. Endothelial cell density and morphology, on the other hand, did not differ significantly between groups. Chronic inflammation's impact on stromal remodelling could explain these corneal alterations. The positive correlation between posterior elevation and the number of flares and duration of inflammation represents the importance of early diagnosis and effective treatment.

Effects of hydrogen peroxide on endothelial function in three-dimensional hydrogel vascular model and regulation mechanism of polar protein Par3

Three-dimensional (3D) cell cultures better reflect the function of endothelial cells (ECs) than two-dimensional (2D) cultures. In recent years, studies have found that ECs cultured in a 3D luminal structure can mimic the biological characteristics and phenotypes of vascular ECs, thus making it more suitable for endothelial dysfunction research. In this study, we used a 3D model and 2D tissue culture polystyrene (TCP) to investigate the effects of cell polarity on hydrogen peroxide (H2O2)-induced endothelial dysfunction and its related mechanisms. We observed the cell morphology, oxidative stress, and barrier and endothelial function of human umbilical vein ECs (HUVECs) in 3D and 2D cultures. We then used Illumina to detect the differentially expressed genes (DEGs) in the 3D-cultured HUVEC with and without H2O2 stimulation, using clusterProfiler for Gene Ontology function enrichment analysis and Kyoto Encyclopaedia of Genes and Genomes pathway enrichment analysis of DEGs. Finally, we explored the role and mechanism of polar protein partitioning defective protein 3 (Par3) in the regulation of ECs. ECs were inoculated into the 3D hydrogel channel; after stimulation with H2O2, the morphology of HUVECs changed, the boundary was blurred, the expression of intercellular junction proteins decreased, and the barrier function of the EC layer was damaged. 3D culture increased the oxidative stress response of cells stimulated by H2O2 compared to 2D TCPs. The polarity-related protein Par3 and cell division control protein 42 were screened using bioinformatics analysis, and western blotting was used to verify the results. Par3 knockdown significantly suppressed claudin1 (CLDN1) and vascular endothelial cadherin. These results suggest that the polar protein Par3 can protect H2O2-induced vascular ECs from damage by regulating CLDN1 and VE-cadherin.

Phosphorylated-S6 Expression in Sturge-Weber Syndrome Brain Tissue

Background: Sturge-Weber Syndrome (SWS) is a rare neurovascular disorder associated with port-wine birthmarks, glaucoma, and abnormal leptomeningeal blood vessels. It is most commonly caused by a R183Q somatic mutation in GNAQ predicted to result in impaired deactivation of the protein Gαq, and downstream pathways such as the Ras-Raf-MEK-ERK and mTOR pathways. The GNAQ mutation is reportedly enriched in endothelial cells. Methods: Using a phosphorylated-S6 (p-S6) antibody, a marker for mTOR pathway activity, we analyzed p-S6 protein expression in human brain tissue samples from SWS subjects and epilepsy controls. Results: SWS brain tissue was more likely to have leptomeningeal endothelial p-S6 antibody staining, compared with epilepsy controls, while the percentage of samples with p-S6 staining in perivascular cells and cortex were not significantly different. Conclusions: Leptomeningeal endothelial mTOR activity may contribute to abnormal endothelial cell morphology and function. This finding implicates mTOR as a potential treatment target for Sturge-Weber Syndrome.

Cold Storage Media versus Optisol-GS in the Preservation of Corneal Quality for Keratoplasty: A Systematic Review

Optisol-GS is the most widely used pharmaceutical composition to preserve corneas for transplantation. This systematic review investigated the effects of different cold corneal storage media (CCSM) compared with Optisol-GS on the quality of stored corneas. The literature was searched throughout May 2022 on six databases and grey literature. Studies including corneas (population) exposed to distinct cold storage media (exposure) and Optisol-GS (comparison) that reported qualitative and/or quantitative parameters of cornea quality (outcome) were included. Methodological quality was assessed using ToxRTool. From 4520 identified studies, fourteen were included according to the eligibility criteria, comprising 769 evaluated cornea samples comparing Optisol-GS with commercial and noncommercial media. All studies showed good methodological quality. Experimental times ranged from 1–28 days, mainly using 4 °C as the preservation temperature. Viable endothelial cell density (ECD) and endothelial cell morphology (EC) were the most assessed parameters. ECD results for Cornisol were higher than Optisol-GS in 10 days (p = 0.049) and favored Cornea ColdTM up to 4 weeks (p &lt; 0.05), which also showed better qualitative results. While the standardization of test protocols could improve comparisons, evidence indicates that most CCSM present similar performances on cornea preservation for transplantation at seven days, while some formulations may increase preservation at extended times.