Human Umbilical Vein Endothelial Cells Research Articles

Synthesis and characterization of activated carbon-supported magnetic nanocomposite (MNPs-OLAC) obtained from okra leaves as a nanocarrier for targeted delivery of morin hydrate.

The method of encapsulating the drug molecule in a carrier, such as a magnetic nanoparticle, is a promising development that has the potential to deliver the medicine to the site where it is intended to be administered. Morin is a pentahydroxyflavone obtained from the leaves, stems, and fruits of various plantsmainly from the Moraceae family exhibiting diverse pharmacological activities such as anti-inflammatory, anti-oxidant, and free radical scavenging and helps treat diseases such as diabetes, myocardial infarction and cancer. In this study, we conducted the synthesis of a nanocomposite with magnetic properties by coating biocompatible activated carbon obtained from okra plant leaves with magnetic nanoparticles. Characterization of the synthesized activated carbon-coated magnetic nanocomposite was confirmed by Fourier transform infrared, scanning electron microscopy, dynamic light scattering, and zeta potential. The cytotoxic effects of the drug-loaded magnetic nanocomposite were examined in HT-29 (Colorectal), MCF-7 (breast), U373 (brain), T98-G (Glioblastoma) cancer cell lines, and human umbilical vein endothelial cells healthy cell line. We studied the loading and release behavior of morin hydrate in the activated carbon-coated magnetic nanocomposite. Activated carbon-coated magnetic nanocomposite carriers can show promising results for the delivery of Morin hydrate drugs to the targeted site.

Synthesis of Carborane–Thiazole Conjugates as Tyrosinase and 11β-Hydroxysteroid Dehydrogenase Inhibitors: Antiproliferative Activity and Molecular Docking Studies

The presented study depicts the synthesis of 11 carborane–thiazole conjugates with anticancer activity, as well as an evaluation of their biological activity as inhibitors of two enzymes: tyrosinase and 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1). The overexpression of tyrosinase results in the intracellular accumulation of melanin and can be observed in melanoma. The overexpression of 11β-HSD1 results in an elevation of glucocorticoid levels and has been associated with the aggravation of metabolic disorders such as type II diabetes mellitus and obesity. Recently, as the comorbidity of melanomas and metabolic disorders is being recognized as an important issue, the search for new therapeutic options has intensified. This study demonstrates that carborane–thiazole derivatives inhibit both enzymes, exerting beneficial effects. The antiproliferative action of all newly synthesized compounds was evaluated using three cancer cell lines, namely A172 (human brain glioblastoma), B16F10 (murine melanoma) and MDA-MB-231 (human breast adenocarcinoma), as well as a healthy control cell line of HUVEC (human umbilical vein endothelial cells). The results show that 9 out of 11 newly synthesized compounds demonstrated similar antiproliferative action against the B16F10 cell line to the reference drug, and three of these compounds surpassed it. To the best of our knowledge, this study is the first to demonstrate dual inhibitory action of carborane–thiazole derivatives against both tyrosinase and 11β-HSD1. Therefore, it represents the first step towards the simultaneous treatment of melanoma and comorbid diseases such as type II diabetes mellitus.

APMCG-1 attenuates ischemic stroke injury by reducing oxidative stress and apoptosis and promoting angiogenesis via activating PI3K/AKT pathway

Ischemic stroke (IS) is a major cause of mortality and morbidity worldwide. Beyond thrombolysis, strategies targeting anti-oxidative apoptosis and angiogenesis are considered prospective therapeutic strategies. Nevertheless, existing natural and clinical remedies have limited efficacy in the management of IS. Moreover, despite their millennial legacy of IS remediation, natural remedies such as ginseng incur high production costs. The novel glycopeptide APMCG-1, extracted from mountain-cultivated ginseng dregs in our previous study, is a potent therapeutic candidate for IS. This study investigated APMCG-1's remedial mechanisms against IS injury using an H2O2-induced oxidative stress paradigm in human umbilical vein endothelial cells (HUVECs) emulating ischemic endothelial cells, in a ponatinib-induced zebrafish IS model, and in rat middle cerebral artery occlusion (MCAO) prototypes. Cellular assays confirmed the proficiency of APMCG-1 in preventing oxidative stress and cell death, fostering regeneration, and facilitating neovascularization within the H2O2-stressed HUVECs framework. Moreover, APMCG-1 augmented hemodynamic velocity, oxidative stress mitigation, apoptosis reduction, and motor enhancement in a zebrafish model of IS. In MCAO rats, APMCG-1 ameliorated neurological deficits and cerebral injury, as evidenced by increased neurological scores and diminished infarct dimensions. In cells and animal models, APMCG-1 activated the PI3K/AKT signaling pathway, modulating factors such as Nrf2, Bcl-2, Caspase 3, eNOS, and VEGFA, thereby ameliorating cellular oxidative distress and catalyzing angiogenesis. Collectively, these results demonstrate the potential protective effects of APMCG-1 in IS pharmacotherapy and its prospective utility as an herbal-derived IS treatment modality.

Zedoarondiol inhibits monocyte adhesion and expression of VCAM and ICAM in endothelial cells induced by oxidative stress

Zedoarondiol, a newly discovered compound derived from the roots of zedoary turmeric, a traditional Chinese herb, has demonstrated potential in reducing inflammation of the vascular endothelium and safeguarding it from harm. Nonetheless, the precise mechanism underlying these effects remains to be elucidated. In this study, we established a model of HUVEC injury induced by hydrogen peroxide. We observed whether Zedoarondiol could reduce the adhesion and transendothelial migration (TEM) of inflammatory cells by inhibiting the expression of VCAM-1 and ICAM-1 in HUVECs. The research findings indicate that utilising Zedoarondiol resulted in a significant reduction in the adhesion rate of THP1 cells to HUVECs, leading to a more condensed cytoskeletal structure. Moreover, Zedoarondiol demonstrated a decrease in NF-κBβ-Ser536 phosphorylation levels in H2O2-stimulated human umbilical vein endothelial cells, resulting in a hindered capacity to bind to target genes like ICAM-1 and VCAM-1, This findings may provide a new pharmacological basis and scientific evidence for Zedoarondiol to slow the atherosclerosis progression by maintaining endothelial function.

Constructing Mg-phytic acid enhanced hydrogel patch with janus structure for antibacterial and angiogenic urethral repair

Due to bacteria in urine and inadequate blood supply, urethral defect repair faces great clinical difficulties. Inspired by the natural structure of the urethra, in this study, we designed a multifunctional hydrogel urethral patch with a Janus structure. The inner layer was a double-network hydrogel of polyvinyl alcohol/carboxymethyl chitosan (PVA/CMCS) enhanced by natural phytic acid (PA) and Mg2+ (Mg2+-PA/PVA/CMCS). The outer layer was composed of poly (L-lactic acid) (PLLA) nanofibers to form a hydrophobic barrier, coupling via a micelle layer to ensure interfacial bonding. The patch exhibited mechanical properties matching soft tissues, with maximum tensile strength and toughness of 1.69±0.13 MPa and 24.7±0.9 MJ/m3, respectively, under the strengthening effect of Mg2+-PA chelation. Therein, the 4 % Mg2+-PA/PVA/CMCS composite patch could maintain unilateral hydrophobicity for at least 14 days and demonstrated an antibacterial rate of 99.0 %±0.1 % against E. coli. In PBS, the patch could deliver Mg2+ for at least 7 days and stabilize the surrounding pH environment for up to 28 days. In vitro cell experiments indicated that the 4 % Mg2+-PA/PVA/CMCS composite patch could promote the human umbilical vein endothelial cell (HUVEC) proliferation and migration, with the angiogenic protein expression enhanced more than 1.2 times compared with the Control group. In conclusion, the patch constructed in this study may provide a new approach to clinical urethral injury repair.

Sera from Rheumatoid Arthritis Patients Induce Oxidative Stress and Pro-Angiogenic and Profibrotic Phenotypes in Human Endothelial Cells.

Background: Rheumatoid arthritis (RA) is a long-term autoimmune condition marked by persistent inflammation of the joints and various systemic complications, including endothelial dysfunction, atherosclerosis, and pulmonary fibrosis. Oxidative stress is a key contributor to the pathogenesis of RA, potentially exacerbating vascular damage and promoting pro-angiogenic and profibrotic processes. Objective: This study aims to investigate the effects of sera from RA patients on human umbilical vein endothelial cells (HUVECs), focusing on the induction of oxidative stress, endothelial cell proliferation, migration, and collagen type I synthesis. Methods: Twenty-eight serum samples were collected from RA patients and healthy donors (HDs). HUVECs were exposed to these sera, and intracellular reactive oxygen species (ROS) levels were fluorescently detected using H2DCF-DA. Cell viability was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cell migration was evaluated through a scratch wound assay, and collagen type I synthesis was measured using a lentiviral vector expressing the green fluorescent protein (GFP) under the control of the human COL1A1 gene promoter. Results: Exposure to RA sera resulted in a significant increase in intracellular ROS levels in HUVECs compared to HD sera, indicating an elevated state of oxidative stress. RA sera also promoted endothelial cell proliferation and migration, suggesting a pro-angiogenic stimulus. Additionally, RA sera significantly increased collagen type I synthesis in HUVECs, implicating a potential role in profibrotic processes associated with RA. Conclusion: The results of this study emphasize the importance of circulating factors in RA sera in promoting oxidative stress, endothelial dysfunction, and pro-angiogenic and profibrotic phenotypes in endothelial cells. These processes may contribute to the vascular and fibrotic complications observed in RA, highlighting the necessity for additional research into focused therapeutic approaches to alleviate these effects.

MicroRNA let-7a regulation of Hantaan virus replication by Targeting FAS Signaling Pathways

Hantaan virus (HTNV) infection in humans can cause hemorrhagic fever and renal syndrome (HFRS). Understanding host responses to HTNV infection is crucial for developing effective disease intervention strategies. Previous RNA-sequencing studies have investigated the role of microRNAs (miRNAs) in the post-transcriptional regulation of host genes in response to HTNV infection. In this study, we demonstrated that HTNV infection induces let-7a expression in human umbilical vein endothelial cells (HUVEC) and that HTNV G protein upregulates the expression of let-7a. miRNA let-7a mimics and inhibitors validated the predicted targets, including cell apoptosis genes (FAS, caspase-8, and caspase-3) and inflammatory factors (IL-6 and its related factors). Modulation of miRNA let-7a levels by miRNA mimics and inhibitors affected HTNV replication, indicating that HTNV modulates host miRNA expression to affect the outcome of the antiviral host response.

Cytotoxic effect of polystyrene nanoplastics in human umbilical vein endothelial cells (HUVECs) and normal rat kidney cells (NRK52E)

Plastics play a crucial role in nearly each aspect of societal production and everyday life, therefore making them one of the most widespread pollutants on a global scale. Because synthetic plastics are not entirely biodegradable, they often remain in the environment and fragment into micro- and nanoplastic particles. The detrimental impacts of nanoplastics on the environment and human health have received substantial worldwide interest in recent years. However, the effects of nanoplastics on human health have not yet been fully explored. Our study aimed to investigate the cytotoxic effects of polystyrene nanoplastics on two distinct mammalian cell lines: normal rat kidney cells (NRK52E) and human umbilical vein endothelial cells (HUVECs). Results showed that polystyrene nanoplastics generate cytotoxicity in both NRK52E and HUVECs in a concentration- and time-dependent manner. Additionally, polystyrene nanoplastics induced pro-oxidant levels (e.g., reactive oxygen species and hydrogen peroxide) and reduced antioxidants (glutathione content and glutathione peroxidase enzyme activity) in both types of cells. We also found that polystyrene nanoplastics cause apoptosis in both NRK52E and HUVECs, as shown by the activation of the caspase-3 enzyme and the loss of mitochondrial membrane potential. Interestingly, it was noticed that the vulnerability of HUVECs cells against polystyrene nanoplastics was a little higher than that of NRK52E cells. Also, the cell toxicity caused by polystyrene nanoplastics in NRK52E and HUVECs was effectively alleviated by co-exposure to a reactive oxygen species inhibitor, N-acetyl-cysteine. This suggests that oxidative stress might be one of the pathways that polystyrene nanoplastics cause cell toxicity. The present work warrants future study to explore the toxicity mechanisms of polystyrene nanoplastics in appropriate in vivo models.

A 3D in vitro model of biphasic calcium phosphate (BCP) scaffold combined with human osteoblasts, osteoclasts, and endothelial cells as a platform to mimic the oral microenvironment for tissue regeneration

Objectives: This study aimed to develop an innovative 3D in vitro model based on the biphasic calcium phosphate (BCP) scaffold combined with human osteoblasts (hOBs), osteoclasts (hOCs), and endothelial cells to evaluate its effects on bone and vascular cells behavior. Methods: To this end, an optimized mixture of hydroxyapatite (HA) and β-tricalcium phosphate (TCP) with a weight ratio of 30/70 was employed to develop a BCP scaffold using the computer-aided design (CAD) approach. The BCP scaffold was combined with primary cultures of hOBs, hOCs and human umbilical vein endothelial cells (HUVECs). Results: Morphometric analyses using scanning electron microscopy (SEM) and X-ray micro-computed tomography, along with biomechanical testing, revealed that BCP scaffold exhibited a regular 3D structure with large interconnected internal pores (700 µm) and high mechanical strength. In terms of biological behavior, after 14 days of tri-culture with hOBs, hMCs and HUVECs, SEM, immunofluorescence, and histological analyses showed that all cell types were viable and adhered well to the entire surface of the scaffold. Interestingly, SEM and energy-dispersive X-ray spectroscopy analyses also revealed on the BCP scaffold the presence of mineralized matrix crystals of Ca, P, O and C within a tissue-like cell layer produced by the interaction of the three cell types. Conclusions: Data confirmed the high performance of the BCP scaffold through biomechanical studies. Notably, for the first time, this study demonstrated the feasibility of combining BCP scaffold with hOBs, hOCs, and HUVEC, which remained viable and maintained their native phenotypes, creating also tissue-like cell layer. Clinical significanceAlthough further investigation is needed, these results underscore the potential to develop a 3D in vitro model that mimics the oral microenvironment, which could be valuable for BTE approaches in in vivo studies.

Long Non-Coding RNA PVT1 Facilitates Radiation-Induced Vascular Endothelial Cell Injury through Sponging MicroRNA-9-5p.

Radiotherapy is a common therapeutic strategy for various solid tumors, with vascular endothelial injury being a common side effect. The study aimed to examine the effect of long non-coding RNA PVT1 on radiation-induced vascular endothelial cell injury, and explore the possible underlying mechanism. Human umbilical vein endothelial cells (HUVECs) were exposed to different doses of X ray to mimic radiation. LncRNA and miRNA levels were detected via qRT-PCR. Interaction between lncRNA and miRNAs was determined through dual-luciferase reporter assay. Statistical processing was conducted using student's t test between two groups and one-way ANOVA among multiple groups, and P < 0.05 means a significant difference. GO and KEGG were performed for the function and pathway enrichment analysis. LncRNA PVT1 elevated along with the increase of radiation dose in HUVECs. Poorly expressed lncRNA PVT1 promotes cell viability and inhibits oxidative stress. PVT1 serves as a competitive endogenous RNA (ceRNA) of miR-9-5p. miR-9-5p inhibitor inverted the influence of PVT1 knockdown on radiation-stimulated cell apoptosis and oxidative stress in HUVECs. KEGG analysis identified significant enrichment of the MAPK signaling pathway among overlapping target genes of miR-9-5p. LncRNA PVT1 knockdown alleviated radiation-induced vascular endothelial injury via sponging miR-9-5p. The underlying mechanism might be probably MAPK signaling-related.

Heparin-binding protein and sepsis-induced coagulopathy: Modulation of coagulation and fibrinolysis via the TGF-β signalling pathway

BackgroundHeparin-binding protein (HBP) levels have been linked to organ failure and may represent an inflammatory biomarker of sepsis. We found disseminated intravascular coagulation (DIC) is associated with higher HBP levels in patients and in in vivo and in vitro models. This prospective, single-center observational study investigated the effects and underlying mechanisms of HBP on the coagulation cascade in sepsis. Methods538 patients with sepsis from June 2016 to December 2019 were enrolled. Mechanisms underlying HBP and the coagulation system were investigated in human umbilical vein endothelial cells (HUVEC) and C57 mice. ResultsIncreased HBP was associated with sepsis-induced DIC. The optimal cutoff value was 37.5 ng/mL (sensitivity: 56 %, specificity: 65 %). Antithrombin-III (AT-III) activity, plasmin-a2 plasmin inhibitor complex (PIC), procalcitonin (PCT), hemoglobin, and HBP ≥37.5 ng/mL were associated with of DIC occurrence. In HUVECs &C57 mice models, Western blotting, qPCR, and immunohistochemistry analysis showed that the binding between HBP and TGF-β receptor 2 (TGFBR2) caused elevation of plasminogen activator inhibitor-1 (PAI-1) levels. Furthermore, we found that mice stimulated with HBP had higher levels of fibrinogen and D-dimer in the blood. HBP treatment caused the accumulation of fibrinogen in mice lung tissue. Treatment with TGFBR2-small interfering RNAs inhibited the effects. ConclusionPatients with sepsis having HBP ≥37.5 ng/mL at admission were more likely to develop DIC. HBP upregulates the expression of fibrinogen and PAI-1 via TGFBR2 and the TGF-β signalling pathway.

Fine-tuning the cytotoxicity profile of N,N,N-trimethyl chitosan through trimethylation, molecular weight, and polyelectrolyte complex nanoparticles.

N,N,N-trimethyl chitosan (TMC) is a promising biopolymer for pharmaceutical applications due to its enhanced solubility and bioadhesive properties, though its cytotoxic limitations necessitate careful modification to ensure safety and efficacy. This study sought to investigate whether nanoparticle (NP) formation could reduce the anticipated cytotoxic effects of TMC, thus improving its applicability across a wider spectrum of pharmaceutical uses. TMC's capability to form NPs with anionic polyelectrolytes led to the application of chondroitin sulfate (ChS) in this study. Five TMC samples, varying in degree of trimethylation (DTM 23, 32, 46, 50 and 99%) and molecular weight (Mw, 66-290kDa) were synthesized, and their biocompatibility with human umbilical vein endothelial cells (HUVECs) was assessed. The results revealed a discernible impact of both DTM and Mw on cell viability, with higher DTM and lower Mw correlating with increased toxicity. Cytotoxicity studies against ovarian cancer cell lines SKOV-3 and OVISE showed a clear indication of a higher cytotoxic effect of TMC samples against cancer cells compared to healthy cells (HUVEC). The cytotoxicity against cancer cells also indicated an optimal DTM for maximum efficacy, deviating from a linear trend. The effects of Mw were cell-dependent, introducing complexity to the observed relationship. Additionally, TMC-ChS NPs were successfully prepared, demonstrating a substantial reduction in cytotoxicity compared to TMC alone in all tested cells. This promising outcome suggests the potential of NP formation to fine-tune the cytotoxicity profile of TMC, paving the way for the development of safer and more effective pharmaceutical formulations.

CLEC14A facilitates angiogenesis and alleviates inflammation in diabetic wound healing

BackgroundDelayed wound healing is a serious complication of diabetic wounds, posing a significant challenge to the treatment of patients with diabetes. Diabetic wound healing is a complex dynamic process involving angiogenesis and inflammatory responses. Currently, there are limited targeted therapies to promote diabetic wound healing. This study aimed to reveal the role of CLEC14A in the process of diabetic wound healing, with the hope of identifying new therapeutic targets to accelerate the healing of diabetic wounds. MethodsIn vivo, diabetic mice were generated by combined streptozotocin (STZ) and high-fat diet treatment. The wound healing model was established in wild-type and Clec14a−/− diabetic mice. The degree of wound healing, as well as angiogenesis and inflammation during the healing process, were evaluated through Hematoxylin and Eosin (H&E) staining, immunohistochemical staining, and immunofluorescence staining. In vitro, the angiogenic activities of Human Umbilical Vein Endothelial Cells (HUVECs) were assessed following treatment with high glucose and adenoviruses overexpressing CLEC14A, using scratch assays and tube formation assays. Interleukin-1β (IL-1β) and Tumor Necrosis Factor-α (TNF-α) were utilized to evaluate the levels of inflammation in HUVECs. ResultsCLEC14A expression was suppressed in diabetic wounds. Deletion of the Clec14a inhibited angiogenesis and activated inflammatory responses in vivo. High-glucose treatment led to decreased CLEC14A expression, impaired angiogenic capacity, and elevated inflammatory levels in vitro. However, adenoviral-mediated overexpression of CLEC14A reversed the response induced by high glucose. ConclusionCLEC14A accelerates diabetic wound healing by promoting angiogenesis and reducing wound inflammation.

3D morphometry of endothelial cells angiogenesis in an extracellular matrix composite hydrogel

Human umbilical vein endothelial cells (HUVECs) play a fundamental role in angiogenesis. Herein, we introduce digital holographic microscopy (DHM) for the 3D quantitative morphological analysis of HUVECs in extracellular matrix (ECM)-based biomaterials as an angiogenesis model. The combination of volumetric information from DHM and the physicochemical and cytobiocompatibility data provided by fluorescence microscopy and cytology offers a comprehensive understanding of the angiogenesis-related parameters of HUVECs within the ECM. DHM enables label-free, non-contact, and non-invasive 3D monitoring of living samples in real time, in a quantitative manner. In this study, the human amniotic membrane (HAM) is decellularized, pulverized, and combined with sodium alginate hydrogel to provide an in vitro substrate for modeling HUVEC angiogenesis. Our results demonstrate that modifying alginate hydrogel with HAM enhances its biofunctionality due to the presence of ECM components. Moreover, the DHM results reveal an increase in its porous properties, which, in turn, aids in interpreting the tubulation results.

Zinc and melatonin mediated antimicrobial, anti-inflammatory, and antioxidant coatings accelerate bone defect repair

Inflammation and bacterial infection are important causes of implant failure, and the development of multifunctional titanium surfaces to address these issues is an effective means of treating infected bone defects. In this study, polyphenols (EGCG) and Zn2+ were first loaded onto the titanium surface to construct an EGCG/Zn2+ polyphenol metal network coating. Then melatonin (MT) was loaded into the EGCG/Zn2+ network structure to prepare the EGCG/Zn2+/MT composite coating. The results proved that the EGCG/Zn2+/MT coating had good mechanical properties, hydrophilicity, corrosion resistance and bioactivity. In vitro, the inhibition rates of EGCG/Zn2+/MT against E. coli and S. aureus were about 97 % and 81 %, respectively. In vitro experiments revealed that EGCG/Zn2+/MT could regulate the polarization of macrophages (RAW264.7) to M2 type, could induce vascularization of human umbilical vein endothelial cells (HUVEC), and could promote the differentiation of pro-osteoblasts (MC3T3-E1) to osteogenesis. Meanwhile, EGCG/Zn2+/MT achieved effective ROS scavenging within HUVEC and MC3T3-E1. In vivo experiments demonstrated that the EGCG/Zn2+/MT coatings possessed favorable biosafety, anti-inflammatory, antimicrobial, and bone repair capabilities. This study provides a simple and versatile strategy for designing multifunctional surfaces with both antimicrobial, anti-inflammatory, antioxidant, angiogenic and osteogenic properties.

Small molecule modulation of insulin receptor-insulin like growth factor-1 receptor heterodimers in human endothelial cells

ObjectivesThe insulin receptor (IR) and insulin like growth factor-1 receptor (IGF-1R) are heterodimers consisting of two extracellular α-subunits and two transmembrane β -subunits. Insulin αβ and insulin like growth factor-1 αβ hemi-receptors can heterodimerize to form hybrids composed of one IR αβ and one IGF-1R αβ. The function of hybrids in the endothelium is unclear. We sought insight by developing a small molecule capable of reducing hybrid formation in endothelial cells. MethodsWe performed a high-throughput small molecule screening, based on a homology model of the apo hybrid structure. Endothelial cells were studied using western blotting and qPCR to determine the effects of small molecules that reduced hybrid formation. ResultsOur studies unveil a first-in-class quinoline-containing heterocyclic small molecule that reduces hybrids by >50% in human umbilical vein endothelial cells (HUVECs) with no effects on IR or IGF-1R. This small molecule reduced expression of the negative regulatory p85α subunit of phosphatidylinositol 3-kinase, increased basal phosphorylation of the downstream target Akt and enhanced insulin/insulin-like growth factor-1 and shear stress-induced serine phosphorylation of Akt. In primary saphenous vein endothelial cells (SVEC) from patients with type 2 diabetes mellitus undergoing coronary artery bypass (CABG) surgery, hybrid receptor expression was greater than in patients without type 2 diabetes mellitus. The small molecule significantly reduced hybrid expression in SVEC from patients with type 2 diabetes mellitus. ConclusionsWe identified a small molecule that decreases the formation of IR: IGF-1R hybrid receptors in human endothelial cells, without significant impact on the overall expression of IR or IGF-1R. In HUVECs, reduction of IR: IGF-1R hybrid receptors leads to an increase in insulin-induced serine phosphorylation of the critical downstream signalling kinase, Akt. The underpinning mechanism appears, at least in part to involve the attenuation of the inhibitory effect of IR: IGF-1R hybrid receptors on PI3-kinase signalling.

Exosomes from adipose-derived stem cells overexpressing microRNA-671–3p promote fat graft angiogenesis and adipogenic differentiation

Exosomes from adipose-derived stem cells (ADSCs) have been demonstrated to benefit angiogenesis, wound healing, and fat grafting. Small noncoding RNAs such as microRNA (miRNA) and circular RNA play critical roles in mediating the function of ADSCs-derived exosomes. However, the underlying mechanisms have not been fully elucidated. In this study, we investigated the function and mechanism of human ADSCs-derived exosomes (hADSCs-Exo) in promoting fat graft angiogenesis and adipogenic differentiation. hADSCs-Exo were isolated and identified, and treatment with hADSCs-Exo enhanced fat graft angiogenesis and adipogenic differentiation in a mouse fat graft implantation model. We found that hADSCs-Exo overexpressed miR-671–3p and promoted human umbilical vein endothelial cell (HUVEC) proliferation, migration, and invasion. Bioinformatics analysis and luciferase reporter assay validated that TMEM127 is a direct target of miR-671–3p. Rescue experiments demonstrated that TMEM127 overexpression partially antagonized the function of hADSCs-Exo in vitro, such as suppressing HUVEC cell proliferation, migration, and invasion. Moreover, TMEM127 overexpression abrogated the function of hADSCs-Exo on fat graft angiogenesis and adipogenic differentiation. Taken together, our findings demonstrate that miR-671–3p-overexpressing exosomes from ADSC promote fat graft angiogenesis and adipogenic differentiation, which highlights the potential of targeting the ADSC-Exosomes-miR-671–3p/TMEM127 axis to improve outcome of fat graft and tissue engineering regenerative medicine.

Selenoprotein M protects cardiac endothelial cell integrity against high-glucose stress via enhancing Parkin-mediated mitophagy

Selenoprotein M (SELENOM) has emerged as a crucial factor in maintaining cellular redox homeostasis and mitigating oxidative damage. This study aims to investigate its protective role in cardiac endothelial cells under hyperglycemic stress, a condition commonly associated with diabetes mellitus and its cardiovascular complications. Diabetic mice model and human umbilical vein endothelial cells (HUVECs) were applied for in vivo and in vitro studies. Results reveal that hyperglycemia significantly downregulates SELENOM expression in both diabetic mouse hearts and primary cultured cardiac endothelial cells. Overexpression of SELENOM in HUVECs mitigated high-glucose-induced FITC-Dextran diffusion and the loss of transendothelial electrical resistance. Additionally, SELENOM overexpression decreased reactive oxygen species (ROS) levels, preserved tight junction protein expression, and maintained cellular structural integrity under hyperglycemic conditions. Furthermore, SELENOM overexpression attenuated high-glucose-induced mitochondrial apoptosis. High-glucose conditions decreased Parkin and increased p62 and Beclin1 expressions. SELENOM overexpression restored Parkin levels and promoted co-localization of LAMP1 and TOMM20. Knockdown of Parkin significantly attenuated these protective effects, suggesting the importance of Parkin in Selenoprotein M-mediated mitophagy. Collectively, these findings suggest that Selenoprotein M enhances Parkin-mediated mitophagy to protect endothelial cells from hyperglycemic stress, offering potential therapeutic insights for diabetic cardiovascular complications.

An open window into endothelial dysfunction: the HUVEC model

Type 2 diabetes mellitus is a prevalent chronic disease globally, with complications such as retinopathy, nephropathy, neuropathy, and cardiovascular disease being major causes of morbidity and mortality. Endothelial dysfunction is a central factor in these complications. Human umbilical vein endothelial cells (HUVEC) serve as a valuable in vitro model within translational research, enabling detailed investigation of endothelial dysfunction and evaluation of therapeutic interventions. HUVEC-based studies bridge basic science and clinical applications, offering a reproducible and accessible model that closely mimics human vascular biology, making them instrumental in developing targeted therapies for cardiovascular complications in diabetes.

Modelling the maternal-fetal interface: An invitro approach to investigate nutrient and drug transport across the human placenta.

The placenta plays a critical role in maternal-fetal nutrient transport and fetal protection against drugs. Creating physiological invitro models to study these processes is crucial, but technically challenging. This study introduces an efficient cell model that mimics the human placental barrier using co-cultures of primary trophoblasts and primary human umbilical vein endothelial cells (HUVEC) on a Transwell®-based system. Monolayer formation was examined over 7 days by determining transepithelial electrical resistance (TEER), permeability of Lucifer yellow (LY) and inulin, localization of transport proteins at the trophoblast membrane (immunofluorescence), and syncytialization markers (RT-qPCR/ELISA). We analysed diffusion-based (caffeine/antipyrine) and transport-based (leucine/Rhodamine-123) processes to study the transfer of physiologically relevant compounds. The latter relies on the adequate localization and function of the amino-acid transporter LAT1 and the drug transporter P-glycoprotein (P-gp) which were studied by immunofluorescence microscopy and application of respective inhibitors (2-Amino-2-norbornanecarboxylic acid (BCH) for LAT1; cyclosporine-A for P-gp). The formation of functional monolayer(s) was confirmed by increasing TEER values, low LY transfer rates, minimal inulin leakage, and appropriate expression/release of syncytialization markers. These results were supported by microscopic monitoring of monolayer formation. LAT1 was identified on the apical and basal sides of the trophoblast monolayer, while P-gp was apically localized. Transport assays confirmed the inhibition of LAT1 by BCH, reducing both intracellular leucine levels and leucine transport to the basal compartment. Inhibiting P-gp with cyclosporine-A increased intracellular Rhodamine-123 concentrations. Our invitro model mimics key aspects of the human placental barrier. It represents a powerful tool to study nutrient and drug transport mechanisms across the placenta, assisting in evaluating safer pregnancy therapies.