Endothelial Cell Morphology Research Articles

Development and Validation of a Flow-Dependent Endothelialized 3D Model of Intracranial Atherosclerotic Disease.

Intracranial atherosclerotic disease (ICAD) is a major cause of stroke globally, with mechanisms presumed to be shared with atherosclerosis in other vascular regions. Due to the scarcity of relevant animal models, testing biological hypotheses specific to ICAD is challenging. We developed a workflow to create patient-specific models of the middle cerebral artery (MCA) from neuroimaging studies, such as CT angiography. These models, which can be endothelialized with human endothelial cells and subjected to flow forces, provide a reproducible ICAD model. Using imaging from the SAMMPRIS clinical trial, we validated this novel model. Computational fluid dynamics flow velocities correlated strongly with particle-derived flow, regardless of stenosis degree. Post-stenotic flow disruption varied with stenosis severity. Single-cell RNA-seq identified flow-dependent endothelial gene expression and specific endothelial subclusters in diseased MCA segments, including upregulated genes linked to atherosclerosis. Confocal microscopy revealed flow-dependent changes in endothelial cell proliferation and morphology in vessel segments related to stenosis. This platform, rooted in the specific anatomy of cerebral circulation, enables detailed modeling of ICAD lesions and pathways. Given the high stroke risk associated with ICAD and the lack of effective treatments, these experimental models are crucial for developing new ICAD-related stroke therapies.

A cotton-derived carbon coating by thermolysis method for enhanced mechanical and bio-compatibility of NiTi alloy

To elevate the performance of NiTi alloy in medical devices, carbon coating derived from natural cotton has been successfully deposited onto the alloy surface using a one-step thermolysis method. Natural cotton soaked in copper sulfate was utilized as carbon raw material, and the deposition process was carried out at low vacuum and temperature requirements. The mechanical and bio-compatibility of the coating performance of resulted carbon layer has been studied comprehensively. The uniformity and mechanical properties of the carbon film prepared at 390 °C, with a holding time of 1–2 h, indicate that it should retain the alloy's excellent mechanical properties and superelasticity. Furthermore, the carbon coating improves the corrosion resistance compared to the raw alloy material. Observations of the proliferation and morphology of adhered human umbilical vein endothelial cells confirm the non-toxic effect of the carbon film. Additionally, the application of the carbon film significantly reduces the contact angle and enhances the adsorption of albumin in blood, suggesting that employing carbon films as bio-active layers is a viable approach to enhance the performance of NiTi-based blood implants and components.

Unfractionated Heparin Enhances Sepsis Prognosis Through Inhibiting Drp1‐Mediated Mitochondrial Quality Imbalance

AbstractUnfractionated heparin (UFH) is commonly used as an anticoagulant in sepsis treatment and has recently been found to have non‐anticoagulant effects, but underlying mechanisms remain unclear. This retrospective clinical data showed that UFH has significant protective effects in sepsis compared to low‐molecular‐weight heparin and enoxaparin, indicating potential benefits of its non‐anticoagulant properties. Recombinant protein chip screening, surface plasmon resonance, and molecular docking data demonstrated that UFH specifically bound to the cytoplasmic Drp1 protein through its zone 2 non‐anticoagulant segment. In‐vitro experiments verified that UFH's specific binding to Drp1 suppressed Drp1 translocation to mitochondria following “sepsis” challenge, thereby improving mitochondrial morphology, function and metabolism in vascular endothelial cells. Consequently, UHF comprehensively protected mitochondrial quality, thus reducing vascular leakage and improving prognosis in a sepsis rat model. These findings highlight the potential of UFH as a sepsis treatment strategy targeting non‐anticoagulation mechanism.

Revisiting Virchow’s triad: exploring the cellular and molecular alterations in cerebral venous congestion

BackgroundCerebral venous thrombosis (CVT) is a rare but serious condition that can lead to significant morbidity and mortality. Virchow’s triad elucidates the role of blood hypercoagulability, blood flow dynamics, and endothelial damage in the pathogenesis of CVT. Cerebral venous congestion (CVC) increases the risk of cerebral venous sinus thrombosis and can lead to recurrent episodes and residual symptoms. However, the precise mechanism by which blood congestion leads to thrombosis remains unclear. Our objective was to investigate the cellular and molecular alterations linked to CVC through analysis of the pathological morphology of venous sinus endothelial cells and transcriptomic profiling.ResultsThis study demonstrated a remarkable correlation between CVC and the phenotypic transformation of endothelial cells from an anticoagulant to a procoagulant state. The findings revealed that cerebral venous stasis results in tortuous dilatation of the venous sinuses, with slow blood flow and elevated pressure in the sinuses and damaged endothelial cells of the retroglenoid and internal jugular vein ligation (JVL) rat model. Mechanistically, analysis of transcriptomic results of cerebral venous sinus endothelial cells showed significant activation of platelet activation, complement and coagulation cascades pathway in the JVL rats. Furthermore, the expression of von Willebrand factor (vWF) and coagulation factor VIII (F8) in the complement and coagulation cascades and Fgg and F2 in the platelet activation was increased in the cerebral venous sinuses of JVL rats than in sham rats, suggesting that endothelial cell injury in the venous sinus induced by CVC has a prothrombotic effect. In addition, endothelial cell damage accelerates coagulation and promotes platelet activation. Significantly, the concentrations of vWF, F2 and F8 in venous sinus blood of patients with internal jugular vein stenosis were higher than in their peripheral blood.ConclusionCollectively, our data suggest that CVC can induce endothelial cell damage, which then exhibits a procoagulant phenotype and ultimately increases the risk of CVT. This research contributes to our understanding of the pathophysiology of CVC associated with procoagulant factors and reexamines the components of Virchow’s triad in the context of CVC.

Impact of oxidized low-density lipoprotein on rat liver sinusoidal endothelial cell morphology and function

Atherogenesis is associated with elevated plasma levels of oxidized low-density lipoproteins (oxLDL). In vivo, oxLDL causes liver endothelial swelling, and disrupts liver sinusoidal endothelial cell (LSECs) fenestrations. We mapped the nanoscale kinetics of these changes in vitro in isolated rat LSECs challenged with oxLDL and monitored viability with endocytosis and cytotoxicity assays. OxLDL disrupted LSEC ultrastructure – increasing oxLDL concentrations and oxidation levels caused sieve plate loss, fenestration fusion, and gap formation. Importantly, these effects were not uniform across all LSECs. LSECs retained the ability to endocytose ligands irrespective of the presence of oxLDL. However, increasing oxidation levels and concentrations of oxLDL inhibited LSEC mediated degradation of endocytosed ligands. Viability was unaffected by any oxLDL challenge. In conclusion, oxLDL disrupts LSEC ultrastructural morphology in vitro but LSECs remain viable and mostly maintain the scavenging function during oxLDL challenge.

Splice Variant of Retinal G-Protein-Coupled Receptor Deletion-Mediated Dysregulation of Autophagy Increases the Susceptibility to Age-Related Macular Degeneration-Like Defects

Introduction: The splice variant of retinal G-protein-coupled receptor deletion (RGR-d) is a persistent component of drusen and may be involved in the pathogenesis of dry age-related macular degeneration (AMD). Increasing evidence has demonstrated the critical role of autophagy in AMD. In this study, we investigated whether RGR-d disrupts autophagy in early dry AMD in vivo and in vitro. Methods: Fundus imaging and fluoroscopy were performed on RGR-d mice created by multiplex gene editing. The retina microstructure was evaluated by performing hematoxylin and eosin (H&E) staining as well as transmission electron microscopy (TEM). Retinal function was assessed by full-field electroretinography (ERG). After lentivirus transfection and stimulation, the permeability, phagocytosis, and tight junctions of ARPE-19 cells were evaluated. Western blotting of ATG5, Beclin-1, LC3II/I, and P62 was performed to detect the changes in autophagy pathways. Results: Atrophy and patchy penetrating hyperfluorescent foci, consistent with early AMD-like defects, were observed in the fundus of 12-month-old RGR-d mice. H&E staining of retinal tissues indicated thinning of each layer of the retinal structure. H&E staining of retinal tissues indicated thinning of each layer of the retinal structure. TEM analysis showed some diffuse granular deposits. And the morphology of choroidal microvascular endothelial cells was degraded and distorted. The morphology of the photoreceptor outer segments showed structural damage, and Bruch’s membrane was thickened. ERG indicated that the photoreceptor of RGR-d mice were dysfunctional. Changes in autophagy-related protein expression were observed in the retinal pigment epithelium and retinal neurepithelium, and autophagy regulation was decreased. Palmitic acid (PA) stimulation caused permeability, phagocytosis, and tight junction dysfunction in cells overexpressing RGR-d. Beclin-1 and LC3II/I expression levels were significantly decreased and that of P62 was elevated in RGR-d cells after PA stimulation. Conclusion: RGR-d disrupts the autophagy pathway, causing the development of an early AMD-like pathophysiology.

AMP dependent protein kinase regulates endothelial heparan sulfate expression in response to an inflammatory stimulus under arterial shear stress

Heparan sulfate (HS) is the most abundant glycosaminoglycan on the vascular endothelium and can regulate endothelial cell morphology and function in response to mechanical stimuli. This study investigated endothelial HS response to an inflammatory stimulus under static and arterial shear stress conditions. Human aortic endothelial cells (HAECs) under static conditions expressed significantly higher HS when treated with an inflammatory stimulus compared to untreated controls. HAECs exposed to an inflammatory stimulus after being conditioned with 10 dyn/cm2 of shear stress for 24 h did not express significantly higher HS compared to untreated controls under flow. To investigate the mechanism underlying this differential endothelial HS expression in response to an inflammatory stimulus under static and shear stress conditions, we hypothesized a shear dependent increase in AMP dependent protein kinase (AMPK) was regulating HS response to the inflammatory stimulus. AMPK inhibition using compound C decreased HAEC HS expression in response to inflammatory stimulus under arterial shear stress, revealing AMPK as a regulator of HS expression. Further investigation is needed to elucidate the mechanistic pathways underlying the interactions between HS and AMPK expression in endothelial cells and how they regulate HAEC inflammatory response.

Comparison of the structural integrity and quality of corneal endothelium stored in organ culture storage medium versus Eusol-C.

In this experimental study, we compared the structural integrity and cell quality of corneal endothelium stored in organ culture medium (OCS) and Eusol-C. The experiment included rabbit and human cornea experiments in vitro. Thirty rabbit corneas and thirty-two human corneas were collected and divided into two groups. All right corneas were allocated in experiment group and left corneas were placed in control group. The corneas in experimental group were stored in OCS at 34°C, and the corneas in control group were stored in Eusol-C at 4°C for 7, 14, 21, 28, and 35days, respectively. Endothelial cell morphology, cell count, and trypan blue staining for viability were assessed before storage (Day 0) and at days 7, 14, 21, 28 and 35. The structural integrity of human corneal endothelial cell was analyzed using immunohistochemistry. The samples of storage solution for microbial culture were collected on the third day and at the end of storage. The results show that no bacterial and fungal infections were found in both groups. After 14days of storage, the morphology of endothelial cell was better in the experimental group than in the control group. The endothelial cell stored in OCS were better than those stored in Eusol-C at the end of storage times, except human cornea 14days storage group. The ZO-1 protein staining showed the typical polygonal morphology of endothelial cell stored in the OCS. Corneal endothelial cells stored in the OCS had better quality up to 28days. It can be applied to Chinese eye banks as a method of corneal preservation.

Vascular endothelial cell morphology and alignment regulate VEGF-induced endothelial nitric oxide synthase activation.

Nitric oxide (NO) production by endothelial nitric oxide synthase (eNOS) inhibits platelet and leukocyte adhesion while promoting vasorelaxation in smooth muscle cells. Dysfunctional regulation of eNOS is a hallmark of various vascular pathologies, notably atherosclerosis, often associated with areas of low shear stress on endothelial cells (ECs). While the link between EC morphology and local hemodynamics is acknowledged, the specific impact of EC morphology on eNOS regulation remains unclear. Morphological differences between elongated, aligned ECs and polygonal, randomly oriented ECs correspond to variations in focal adhesion and cytoskeletal organization, suggesting differing levels of cytoskeletal prestress. However, the functional outcomes of cytoskeletal prestress, particularly in the absence of shear stress, are not extensively studied in ECs. Some evidence suggests that elongated ECs exhibit decreased immunogenicity and enhanced NO production. This study aims to elucidate the signaling pathways governing VEGF-stimulated eNOS regulation in the aligned EC phenotype characterized by elongated and aligned cells within a monolayer. Using anisotropic topographic cues, bovine aortic endothelial cells (BAECs) were elongated and aligned, followed by VEGF treatment in the presence or absence of cytoskeletal tension inhibitors. Phosphorylation of eNOS ser1179, AKT ser437 and FAK Tyr397 in response to VEGF challenge were significantly heightened in aligned ECs compared to unaligned ECs. Moreover this response proved to be robustly tied to cytoskeletal tension as evinced by the abrogation of responses in the presence of the myosin II ATPase inhibitor, blebbistatin. Notably, this work demonstrates for the first time the reliance on FAK phosphorylation in VEGF-mediated eNOS activation and the comparatively greater contribution of the cytoskeletal machinery in propagating VEGF-eNOS signaling in aligned and elongated ECs. This research underscores the importance of utilizing appropriate vascular models in drug development and sheds light on potential mechanisms underlying vascular function and pathology that can help inform vascular graft design.

Changes in Corneal Endothelial Cells and Central Corneal Thickness in Pseudoexfoliation Syndrome

Purpose: The aim of the research is to investigate the impact of pseudoexfoliation syndrome (PEX) on corneal endothelial cells and central corneal thickness. Methods: In this analytic case-control study, the research sample included 60 patients (120 eyes), which was divided into two groups: the PEX group, 30 patients (60 eyes) obtained inclusion criteria and the control group, 30 patients (60 eyes) for result comparison, from patients visiting the ophthalmic clinic at Tishreen University Hospital. PEX degree was assessed using a slit lamp and the morphology of endothelial cells: endothelial cell density (ECD), coefficient of variation (COV) , percentage of hexagonal cells (EX%), average cell area (Avg.Cell Area) and central corneal thickness (CCT) were assessed using a specular microscope. Results: The average of ECD was lower in the PEX group compared with the control group (p-value = 0.0001). The average of EX% was lower in the PEX group compared with the control group (p-value = 0.0001). No statistically significant differences were observed between the research groups in COV values (p-value = 0.9). The Avg.Cell Area was higher in the PEX group compared with the control group (p-value = 0.003). The average of CCT was lower in the PEX group compared with the control group (p-value = 0.04). Conclusion: Corneal endothelial cells morphology and central corneal thickness are affected in eyes with PEX syndrome, and these changes increase with the severity of PEX.

Improved Composite Hydrogel for Bioengineered Tracheal Graft Demonstrates Effective Early Angiogenesis.

Background/Objectives: Collagen-agarose hydrogel blends currently used in tracheal graft bioengineering contain relatively high concentrations of collagen to withstand mechanical stresses associated with native trachea function (e.g., breathing). Unfortunately, the high collagen content restricts effective cell infiltration into the hydrogel. In this study, we created an improved hydrogel blend with lower concentrations of collagen (<5 mg/mL) and characterized its capacity for fibroblast invasion and angiogenesis. Methods: Four collagen-agarose hydrogel blends were created: 1 mg/mL type 1 collagen (T1C) and 0.25% agarose, 1 mg/mL T1C and 0.125% agarose, 2 mg/mL T1C and 0.25% agarose, and 2 mg/mL T1C and 0.125% agarose. The hydrogel surface was seeded with fibroblasts, while both endothelial cells and fibroblasts (3:1 ratio) were mixed within the hydrogel matrix. We assessed early angiogenesis by observing fibroblast migration and endothelial cell morphology (elongation and branching) at 7 days. In addition, we performed immunostaining for alpha-smooth muscle actin (aSMA) and explored the gene expression of various angiogenic markers (including vascular endothelial growth factor; VEGF). Results: Gels with lower agarose concentrations (0.125%) with 1 or 2 mg/mL T1C were more effective in allowing early attachment and migration of surface-applied fibroblasts compared to gels with higher (0.25%) agarose concentrations. The low-agarose gels also allowed cells to quickly adopt a spread morphology and self-assemble into elongated structures indicative of early angiogenesis, while demonstrating positive immunostaining for aSMA and increased gene expression of VEGF by day 7. Conclusions: Hydrogel blends with collagen and low agarose concentrations may be effective in allowing early cellular infiltration and angiogenesis, making such gels a suitable cell substrate for use in the development of composite bioengineered tracheal grafts. The collagen-agarose hydrogel blend is meant to be cast around a three-dimensional (3D) printed polycaprolactone support structure and wrapped in porcine small intestine submucosa ECM to create an off-the-shelf bioengineered tracheal implant.

Catalpol inhibits HHcy-induced EndMT in endothelial cells by modulating ROS/NF-κB signaling

BackgroundHyperhomocysteinemia (HHcy) is an independent risk factor for atherosclerosis (AS). Endothelial mesenchymal transition (EndMT) refers to the process in which endothelial cells lose endothelial cell morphology and characteristic gene expression, and acquire phenotypic characteristics and gene expression related to mesenchymal cells. Numerous studies have confirmed that EndMT is involved in the formation of atherosclerosis. Catalpol is one of the active components of Rehmannia, which has antioxidant, anti-inflammatory, anti-tumor, neuroprotective and other biological activities. Studies have shown that catalpol can reduce atherosclerotic plaque induced by high sugar or fat. However, the effect of catalpol on HHCY-induced EndMT is unclear.Methods and resultsIn vitro HHcy-treated primary human umbilical vein endothelial cells (HUVECs) were used to construct a cell model, and the antioxidants N-acetylcysteine (NAC) and catalase alcohol were administered. In vivo C57BL/6N mice were given a diet fed with 4.4% high methionine chow to construct a HHcy mice model and were treated with catalpol. The results showed that hhcy could induce morphological transformation of endothelial cells into mesenchymal cells, increase intracellular ROS content, up-regulate α-SMA, N-cadherin, p-p65 protein expression, down-regulate VE-cadherin, CD31 protein expression, induce pathological changes of aortic root endothelium, and increase aortic endothelial ROS content. Catalpol reversed these hhcy induced outcomes.ConclusionsCatalpol inhibits HHcy-induced EndMT, and the underlying mechanism may be related to the ROS/NF-κB signaling pathway. Catalpol may be a potential drug for the treatment of HHcy-related AS.

Corneal Endothelial Cell Density and Morphology in Healthy Libyan Eyes

Background Endothelial cell density and morphology vary across ethnic groups. Aim This article investigates the corneal endothelial cell density and morphology in healthy Libyan eyes. Methods A 3-month cross-sectional observational study was conducted at Benghazi Teaching Eye Hospital, involving 198 eyes of 100 healthy Libyan participants. The noncontact Topcon specular microscope (SP-1P model) was used to assess the following parameters: the mean central corneal thickness (CCT), the mean cell density (MCD), the mean coefficient of variation (CV%), and the mean percentage of the hexagonal cell (Hex [%]). The variables were analyzed in relation to age and gender using the Statistical Package for the Social Sciences (SPSS version 25.0) Results The mean age of participants in this study was 47.4 ± 13.8 years (range 21–75 years). The mean CCT was 516.45 ± 43.04 μm, the MCD was 2664.30 ± 371.26 cells/mm2, the mean CV% was 32.3% ± 3.7, and the mean Hex (%) was 52.8% ± 9.6. There was no statistical difference in the age, CCT, and MCD across genders. Whereas CV (%) and Hex (%) showed significant gender differences (p &lt; 0.01 for both). There was a significant negative weak correlation between CCT (r = –0.10) and age, as well as a significant negative moderate correlation between MCD and Hex (%) with age (r = –0.36 and r = –0.31, respectively). CV% exhibited a significant, moderately positive association with age (r = 0.35). The higher endothelial cell loss rate of 8.4% was in the third decade of life whereas other age groups ranged between 1.1 and 2.7%. Conclusion The normative data for the endothelium of Libyan eyes are reported, which can be used as a baseline for future studies.

Novel mathematical approach to accurately quantify 3D endothelial cell morphology and vessel geometry based on fluorescently marked endothelial cell contours: Application to the dorsal aorta of wild-type and Endoglin-deficient zebrafish embryos.

Endothelial cells, which line the lumen of blood vessels, locally sense and respond to blood flow. In response to altered blood flow dynamics during early embryonic development, these cells undergo shape changes that directly affect vessel geometry: In the dorsal aorta of zebrafish embryos, elongation of endothelial cells in the direction of flow between 48 and 72 hours post fertilization (hpf) reduces the vessel's diameter. This remodeling process requires Endoglin; excessive endothelial cell growth in the protein's absence results in vessel diameter increases. To understand how these changes in vessel geometry emerge from morphological changes of individual endothelial cells, we developed a novel mathematical approach that allows 3D reconstruction and quantification of both dorsal aorta geometry and endothelial cell surface morphology. Based on fluorescently marked endothelial cell contours, we inferred cross-sections of the dorsal aorta that accounted for dorsal flattening of the vessel. By projection of endothelial cell contours onto the estimated cross-sections and subsequent triangulation, we finally reconstructed 3D surfaces of the individual cells. By simultaneously reconstructing vessel cross-sections and cell surfaces, we found in an exploratory analysis that morphology varied between endothelial cells located in different sectors of the dorsal aorta in both wild-type and Endoglin-deficient zebrafish embryos: In wild-types, ventral endothelial cells were smaller and more elongated in flow direction than dorsal endothelial cells at both 48 hpf and 72 hpf. Although dorsal and ventral endothelial cells in Endoglin-deficient embryos had similar sizes at 48 hpf, dorsal endothelial cells were much larger at 72 hpf. In Endoglin-deficient embryos, elongation in flow direction increased between 48 hpf and 72 hpf in ventral endothelial cells but hardly changed in dorsal endothelial cells. Hereby, we provide evidence that dorsal endothelial cells contribute most to the disparate changes in dorsal aorta diameter in wild-type and Endoglin-deficient embryos between 48 hpf and 72 hpf.

Increased basal fibronectin is sufficient to promote excess endothelial cell matrix assembly causing localized barrier dysfunction.

Endothelial cell behavior is regulated by subendothelial extracellular matrix (ECM). The ECM protein fibronectin (FN) is rare in healthy blood vessels but accumulates in disease accompanied by endothelial dysfunctions. Here, we report that excess assembly of FN disrupts key endothelial functions. We mimicked increased FN expression as in diseased stroma by providing exogenous FN basally in a Transwell insert and found dose-dependent upregulation of subendothelial FN matrix assembly. Taking advantage of discontinuous matrix assembly by endothelial cells, we show correlations between regional increases in FN matrix and disruptions in endothelial cell morphology, VE-cadherin junctions, and the cell cycle, all of which were not changed in FN-deficient regions of the monolayer. These changes affected endothelial barrier function with increased monolayer permeability exposing basal regions of high FN matrix and permitting FN-dependent adhesion of MDA-MB-231 tumor cells from the apical side of the monolayer. FN matrix accumulation was quick and increases in FN matrix preceded all other changes in the endothelium. Therefore, subendothelial accumulation of FN matrix is a cause, not an effect, of endothelial monolayer disorganization and leakiness. Regulating FN accumulation in the subendothelial space could be an important target for controlling progression of fibrosis and related diseases.

Effect of Intravitreal Injection of Ranibizumab (Lucentis®) on Corneal Endothelial Cell Density and Morphology

Abstract Background VEGF has an essential role in initiating wound and inflammation-related neovascularization. Several studies have demonstrated that VEGF is an important mediator of blood– retinal-barrier breakdown, leading to fluid leakage below the macula and inducing the development of macular edema and choroidal neovascular membranes. Anti-VEGF agents interfere with its receptor binding, thus inhibiting VEGF’s signal, which results in inhibition of abnormal blood vessel formation and decreasing vascular permeability. Aim of the Work This study aimed is to report early changes in the corneal endothelial cell density (ECD) and morphology in patients undergoing intravitreal anti-VEGF injections (ranibizumab) for treatment of macular edema. Patients and Methods This prospective study included 22 eyes of 22 DME patients indicated for IVI of Ranibizumab, with mean age 57.1±7.4 years, ranging from 42.0 years to 70.0 years. The majority of them were females (77.3%). Results Cell density (endothelial cell count) at baseline (2601.8±220.0) didn’t show statistically significant difference than that at one month (2598.9±198.1) after IVI. Coefficient variant was not significantly changed at month-1 as compared to baseline. Coefficient variant at baseline was 29.8±4.0, ranging from 22.0 to 39.0. and at one month was 31.3±5.4, ranging from 21.0 to 48.0. Hexagonal cells (HEX) were not significantly changed at one month as compared to baseline. Hexagonal cells at baseline were 68.9±3.2, ranging from 63.0 to 75.0 and after one month, it was 67.8±4.1, ranging from 60.0 to 76.0. Pachymetry was not significantly changed at one month as compared to baseline. The corneal thickness (pachymetry) at baseline was 538.8±40.1 ranging from 448.0–587.0 to 447.0–597.0. after one month of IV ranibizumab it was 536.3±42.1 0.183. Conclusion A single IV injection of Ranibizumab in phakic and otherwise normal eyes with DME showed no change in ECD and pachymetry but a statistically non-significant increase in CV and decrease in HEX 1-month after injection.

Tripterygium drug-loaded liposome alleviates renal function by promoting vascularization and inhibiting fibrosis.

Introduction: Tripterygium species have been traditionally used in Chinese medicine for treating various conditions. The aim of the study was to construct a drug-modified renal infarction targeting liposome (rTor-LIP) containing Tripterygium in order to improve the therapeutic effect on renal injury. Methods: rTor-LIP was prepared using the extruder method containing Tripterygium solution. The preparation was characterized by transmission electron microscopy, Marvin laser particle size analyzer, and Western blotting. In vitro experiments were conducted to verify the biocompatibility of rTor-LIP, and in vivo experiments were conducted to verify the therapeutic effect of rTor- LIP on renal injury. Results and discussion: The surface of rTor-LIP was regular and oval. In vitro results showed that after co-incubation with rTor-LIP, endothelial cells did not show significant apoptosis, and there were no significant abnormalities in the mitochondrial metabolism. The in vivo results showed that the morphology of endothelial cells in the rTor-LIP group was uniform and the cytoplasmic striations were clear, but the local striations had disappeared. Thus, rTor-LIP nano-targeted liposomes can effectively target hypoxic kidney tissue, providing a new idea for the treatment of renal infarction.

Correlations of Corneal Endothelial Morphology and Corneal Thickness With Anterior Segment Parameters in Healthy Individuals.

The purpose of this study was to investigate the associations between central corneal endothelial cell density (ECD), endothelial morphology, and corneal thickness (central corneal thickness) with the anterior chamber depth, corneal volume (CV), white-to-white (WTW) distance, mean anterior chamber angle (CAmean), and gender in healthy individuals. This observational study included 136 healthy volunteers. The ECD, coefficient of variation of cell area, and hexagonal cell appearance ratio (%Hex) were measured by means of specular microscopy. The central corneal thickness, anterior chamber depth, CV, WTW distance, and the angle width of 12 points were taken by the Pentacam HR Scheimpflug anterior segment imaging. The arithmetical mean of the 12 points was considered as the CAmean. We used mixed effect linear regression model for the statistical analysis of the data. ECD was positively correlated with CV ( P = 0.028), while after adjusting for age, it was negatively correlated with age ( P < 0.001). Coefficient of variation of cell area was positively correlated with CAmean ( P = 0.036), while after adjusting for age, it was positively correlated with age ( P < 0.001) and CAmean ( P = 0.005). Hex was negatively correlated with WTW ( P = 0.023) and CAmean ( P = 0.025), and after adjusting for age, this correlation remained the same ( P = 0.029 when correlated with WTW and P = 0.035 with CAmean). There were significant changes in the morphology of the corneal endothelial cells in eyes with wider anterior chamber angle. Greater pleomorphism and polymegethism of the corneal endothelium was observed in healthy patients with wider CAmean. Deepening of the anterior chamber as myopia progresses could render the corneal endothelium more fragile and susceptible to mechanical stress, which is an area worthy of further study.

Dynamics of endothelial cells migration in nature-mimicking blood vessels

Endothelial cells (ECs) migration is a crucial early step in vascular repair and tissue neovascularization. While extensive research has elucidated the biochemical drivers of endothelial motility, the impact of biophysical cues, including vessel geometry and topography, remains unclear. Herein, we present a novel approach to reconstruct 3D self-assembly blood vessels-on-a-chip that accurately replicates real vessel geometry and topography, surpassing conventional 2D flat tube formation models. This vessels-on-a-chip system enables real-time monitoring of vasculogenesis and ECs migration at high spatiotemporal resolution. Our findings reveal that ECs exhibit increased migration speed and directionality in response to narrower vessel geometries, transitioning from a rounded to a polarized morphology. These observations underscore the critical influence of vessel size in regulating ECs migration and morphology. Overall, our study highlights the importance of biophysical factors in shaping ECs behavior, emphasizing the need to consider such factors in future studies of endothelial function and vessel biology.

Corneal Endothelial Morphology and Ocular Biometric Indexes in Premature Children With and Without Retinopathy of Prematurity.

The purpose of this study was to analyze human corneal endothelial cells (HCECs) morphology and ocular biometrics in premature (PM) children with or without retinopathy of prematurity (ROP). Retrospective data on patient demographics, HCECs status, and ocular biometrics with at least 2 visits between 2016 and 2021 were reviewed. The main outcomes were endothelial cell density (ECD), coefficient of variation (CV), hexagonal cell ratio (HEX), central corneal thickness (CCT), axial length, anterior chamber depth, keratometry, corneal diameter, pupil diameter, and refraction status. Generalized estimating equation was used to evaluate the differences between PM no-ROP and ROP groups. We also analyzed the trend of ECD, CV, HEX, and CCT change with age between groups. The study included 173 PM patients without ROP and 139 patients with ROP. A total of 666 and 544 measurements were recorded in the PM no-ROP and ROP groups, respectively. The ROP group had higher spherical power, myopic spherical equivalent (SE), and steeper steep keratometry (K; P < 0.05). The ROP group had higher CV (P = 0.0144), lower HEX (P = 0.0012) and thicker CCT (P = 0.0035). In the HCECs parameters, the ROP group had slower ECD decrement (P < 0.0001), faster CV decrement (P = 0.0060), and faster HEX increment (P = 0.0001). A difference in corneal morphology changes between the ROP and PM no-ROP groups were prominent in patients with lower gestational age (GA) in the subgroup analysis. Worse HCECs morphology and higher myopic status were initially observed in patients with prior ROP but not in PM patients with no-ROP. ECD and HCECs morphology improved with age, especially in patients with low GA.