Human Umbilical Cord Vein Research Articles

Atherosclerosis on a Chip: A 3-Dimensional Microfluidic Model of Early Arterial Events in Human Plaques.

Realistic reconstruction of the in vivo human atherosclerotic environment requires the coculture of different cell types arranged in atherosclerotic vessel-like structures with exposure to flow and circulating cells, presenting challenges for disease modeling. This study aimed to develop a 3-dimensional tubular microfluidic model with quadruple coculture of human aortic smooth muscle cells, human umbilical cord vein endothelial cells, and foam cells to recreate a complex human atherosclerotic vessel in vitro to study the effects of flow and circulating immune cells. We developed a coculture protocol utilizing BFP (blue fluorescent protein)-labeled human aortic smooth muscle cells, GFP (green fluorescent protein)-labeled human umbilical cord vein endothelial cells, and THP-1 macrophage-derived, Dil-labeled oxidized LDL (low-density lipoprotein) foam cells within a fibrinogen/collagen I-based 3-dimensional ECM (extracellular matrix). Perfusion experiments were conducted for 24 hours on both atherosclerotic vessels and healthy vessels (BFP-labeled human aortic smooth muscle cells and GFP-labeled human umbilical cord vein endothelial cells without foam cells). Additionally, perfusion with circulating THP-1 monocytes was performed to observe cell extravasation and recruitment. The resulting vessels displayed early lesion morphology, with a layered composition including an endothelium and media, and foam cells accumulating in the subendothelial space. The layered wall composition of both atherosclerotic and healthy vessels remained stable under perfusion. Circulating THP-1 monocytes demonstrated cell extravasation into the atherosclerotic vessel wall and recruitment to the foam cell core. The qPCR analysis indicated increased expression of atherosclerosis markers in the atherosclerotic vessels and adaptation of vascular smooth muscle cell migration in response to flow and the plaque microenvironment, compared with control vessels. The human 3-dimensional atherosclerosis model demonstrated stability under perfusion and allowed for the observation of immune cell behavior, providing a valuable tool for the atherosclerosis research field.

Vasorelaxant effect of phenylpropanoids: Methyl eugenol and eugenol in human umbilical cord vein

Methyl-eugenol (ME) and eugenol (EUG) are phenylpropanoids with vasodilatory effects. While EUG's vasorelaxant effect in human umbilical artery (HUA) is known, their action in veins is unclear. This study aimed to evaluate ME and EUG in human umbilical vein (HUV). Isolated HUV underwent tension recordings. ME and EUG caused 100 % relaxation in HUV, with EC50 values corresponding to: 174.3 ± 7.3 and 217.3 ± 6.2 µM for ME and EUG respectively in presence of K+; 362.3 ± 5.4 and 227.7 ± 4.9 µM for ME and EUG respectively and in presence of serotonin (5-HT). It was observed that in presence of BaCl2 and CaCl2 evoked contractions, ME (800 and 1000 µM) and EUG (1000 and 1400 µM) prevent the contractions. In presence of K+ channel blockers it was observed that ME promoted relaxation compared to its control, except in presence of 4-AP, suggesting a possible Ca2+-dependent K+ channel activation for this molecule; EUG increased all EC50 in presence of the K+ blockers except in presence of TEA 1 mM. Greater pharmacological potency was observed for ME. This study highlights natural substances' effects on HUV contractile parameters, suggesting ME and EUG as potential vasodilators in maintaining fetal oxygenation and venous flow during gestational hypertensive syndromes.

Orthotopic transplantation of the bioengineered lung using a mouse-scale perfusion-based bioreactor and human primary endothelial cells

Whole lung engineering and the transplantation of its products is an ambitious goal and ultimately a viable solution for alleviating the donor-shortage crisis for lung transplants. There are several limitations currently impeding progress in the field with a major obstacle being efficient revascularization of decellularized scaffolds, which requires an extremely large number of cells when using larger pre-clinical animal models. Here, we developed a simple but effective experimental pulmonary bioengineering platform by utilizing the lung as a scaffold. Revascularization of pulmonary vasculature using human umbilical cord vein endothelial cells was feasible using a novel in-house developed perfusion-based bioreactor. The endothelial lumens formed in the peripheral alveolar area were confirmed using a transmission electron microscope. The quality of engineered lung vasculature was evaluated using box-counting analysis of histological images. The engineered mouse lungs were successfully transplanted into the orthotopic thoracic cavity. The engineered vasculature in the lung scaffold showed blood perfusion after transplantation without significant hemorrhage. The mouse-based lung bioengineering system can be utilized as an efficient ex-vivo screening platform for lung tissue engineering.

Chitosan-Rapamycin Carbon Dots Alleviate Glaucomatous Retinal Injury by Inducing Autophagy to Promote M2 Microglial Polarization.

Glaucoma is a prevalent cause of irreversible vision impairment, characterized by progressive retinal ganglion cells (RGCs) loss, with no currently available effective treatment. Rapamycin (RAPA), an autophagy inducer, has been reported to treat glaucoma in rodent models by promoting RGC survival, but its limited water solubility, systemic toxicity, and pre-treatment requirements hinder its potential clinical applications. Chitosan (CS)-RAPA carbon dot (CRCD) was synthesized via hydrothermal carbonization of CS and RAPA and characterized by transmission electron microscopy, Fourier transform infrared spectra, and proton nuclear magnetic resonance. In vitro assays on human umbilical cord vein endothelial and rat retinal cell line examined its biocompatibility and anti-oxidative capabilities, while lipopolysaccharide-stimulated murine microglia (BV2) assays measured its effects on microglial polarization. In vivo, using a mouse retinal ischemia/reperfusion (I/R) model by acute intraocular pressure elevation, the effects of CRCD on visual function, RGC apoptosis, oxidative stress, and M2 microglial polarization were examined. CRCD exhibited good water solubility and anti-oxidative capabilities, in the form of free radical scavenging. In vitro, CRCD was bio-compatible and lowered oxidative stress, which was also found in vivo in the retinal I/R model. Additionally, both in vitro with lipopolysaccharide-stimulated BV2 cells and in vivo with the I/R model, CRCD was able to promote M2 microglial polarization by activating autophagy, which, in turn, down-regulated pro-inflammatory cytokines, such as IL-1β and TNF-α, as well as up-regulated anti-inflammatory cytokines, such as IL-4 and TGF-β. All these anti-oxidative and anti-inflammatory effects ultimately aided in preserving RGCs, and subsequently, improved visual function. CRCD could serve as a potential novel treatment strategy for glaucoma, via incorporating RAPA into CDs, in turn not only mitigating its toxic side effects but also enhancing its therapeutic efficacy.

Development of a Tri-Layered Vascular Construct and In Vitro Evaluation of Endothelization.

Advances in the development of vascular substitutes for small-sized arteries are ongoing because the present grafts do not entirely meet the requirements of native equivalents and are suboptimal in clinical performance. This study aims to develop a tri-layered vascular construct mimicking natural tissue using polyester blends and to investigate its endothelization through in vitro studies as a potential small-caliber vascular graft. The innermost layer is obtained by dip coating as a tubular porous film with a lumen diameter of 3mm and a pore size of ≤8µm. Circumferentially aligned electrospun fiber (diameter 100-800nm) with a deviation angle of 15° are deposited over the porous film forming the intermediate layer. The random electrospun fibers (diameter 100-1100nm) deviating at different angles are wrapped as the outermost layer. The mechanical properties of the tri-layered vascular construct are determined to be 44.80±14.80MPa for Young's modulus and 4.25±0.75MPa for ultimate tensile strength. MTS and cell behavior studies show that the isolated human umbilical cord vein endothelial cells proliferate and line the lumen of the vascular substitute. The vascular construct developed, with its biomimetic architecture, mechanical features, size, and endothelization, can be tested with in vivo studies.

Establishment of a 3D multicellular placental microtissues for investigating the effect of antidepressant vortioxetine

The placenta is a unique organ with an active metabolism and dynamically changing physiology throughout pregnancy. It is difficult to elucidate the structure of cell-cell and cell-extracellular matrix interactions of the placenta in in vivo studies due to interspecies differences and ethical constraints. In this study, human umbilical cord vein cells (HUVEC) and human placental choriocarcinoma cells (BeWo) were co-cultured for the first time to form spheroids (microtissues) on a three-dimensional (3D) Petri Dish® mold and compared with a traditional two-dimensional (2D) system. Vortioxetine is an antidepressant with a lack of literature on its use in pregnancy in established cultures, the toxicity of vortioxetine was studied to investigate the response of spheroids representing placental tissue. Spheroids were characterised by morphology and exposed to vortioxetine. Cell viability and barrier integrity were then measured. Intercellular junctions and the localisation of serotonin transporter (SERT) proteins were demonstrated by immunofluorescence (IF) staining in BeWo cells. Human chorionic gonadotropin (beta-hCG) hormone levels were also measured. In the 3D system, cell viability and hormone production were higher than in the 2D system. It was observed that the barrier structure was impaired, the structure of intracellular skeletal elements was altered and SERT expression decreased depending on vortioxetine exposure. These results demonstrate that the multicellular microtissue placenta model can be used to obtain results that more closely resemble in vivo toxicity studies of various xenobiotics than other 2D and mono-culture spheroid models in the literature. It also describes the use of 3D models for soft tissues other than the placenta.

Effect of preconditioning of human umbilical cord mesenchymal stem cells with hydrogen peroxide on the therapeutic potential of MSCs in the cyclophosphamide -induced premature ovarian failure mice model.

Exposure of stem cells to sublethal levels of hydrogen peroxide (H2O2) can prevent oxidative stress-induced apoptosis. In the present study, the effects of H2O2 preconditioning on the therapeutic potential of human umbilical vein cord mesenchymal stem cells (hUCV-MSCs) were evaluated in a murine model of premature ovarian failure. Mature mice were divided into 4 groups, and 10 mice were incorporated into each group. The control (Ctrl) group received phosphate buffered saline (PBS) intraperitoneal (IP), and the CTX group was injected IP with cyclophosphamide (CTX). The CTX+MSC group after receiving CTX was injected with a single dose of hUCV-MSCs labeled with CM-DiI intravenously (IV), whereas the CTX+preMSCs group after CTX injection received preconditioned MSCs with H2O2 IV. Seven days later, the mice were euthanized, and their ovaries were removed for histological studies such as H&E staining and the TUNEL assay. Furthermore, the numbers of CM-DiI-labeled hUCV-MSCs in the different regions of the ovary were calculated. FSH and estradiol values in the serum were measured. Our studies showed that CTX caused degenerative changes and follicular loss in the ovary. The number of follicles in the CTX+MSCs and CTX+PreMSCs groups was significantly higher compared to the CTX group. In addition, in the CTX+PreMSCs group, the numbers of different types of follicles were higher than in the CTX-MSC group. Immunohistochemical studies in the CTX+MSCs and CTX+PreMSCs groups showed little evidence of TUNEL positivity compared with the CTX group. Moreover, the apoptotic index decreased in the CTX+PreMSCs group compared to the CTX+MSCs group. Moreover, CM-DiI-labeled MSCs in the ovary in the CTX+pre-MSCs group were higher than in the CTX+MSCs group. Our experiment offers preconditioning as an effective strategy in stem cell therapy to potentiate MSCs' therapeutic efficacy in ovarian function failure.

PHEMA-based cryogels as 3D placental scaffolds

The placental barrier regulates the passage of nutrients, waste, drugs, and chemicals between maternal and fetal tissues. Mono and co-culture systems were constituted with human placental choriocarcinoma cells (BeWo) and human umbilical cord vein cells (HUVEC) on biocompatible poly (2-hydroxyethyl methacrylate) (PHEMA)-based cryogels. PHEMA cryogels were supported with extracellular matrix (ECM) elements. According to the Fourier transform infrared spectrometry-Attenuated Total Reflectance (FTIR-ATR) results, it was confirmed that gelatin, folic acid (FA), fibronectin (FN), and collagen type IV (COL Type IV) were successfully incorporated into the structure of cryogels. The mean pore size of the cryogels confirmed by micro-computed tomography analysis was in the range of 15–195 µm. The antidepressant active ingredient Vortioxetine was investigated by 3-(4,5-dimethylthiazol2-yl)− 2,5-diphenyltetrazolium-bromide (MTT) viability analysis in the constituted cryogel co-culture system. The cryogels did not cause any toxic effect on the cells. High doses of Vortioxetine showed reduced cell viability in monocultures, but not the expected reduction in cell viability at the same doses of co-culture. The highest amount of human chorionic gonadotropin hormone was in the PHEMA+COL Type IV+FA group. There were no significant changes in matrix metalloproteinase (MMP) 2 and 9 protein levels due to Vortioxetine dose administration.

3D bioprinting microgels to construct implantable vascular tissue

Engineered implantable functional thick tissues require hierarchical vasculatures within cell-laden hydrogel that can mechanically withstand the shear stress from perfusion and facilitate angiogenesis for nutrient transfer. Yet current extrusion-based 3D printing strategies are unable to recapitulate hierarchical networks, highlighting the need for bioinks with tunable properties. Here, we introduce an approach whereby crosslinkable microgels enhance mechanical stability and induce spontaneous microvascular networks comprised of human umbilical cord vein endothelial cells (HUVECs) in a soft gelatin methacryoyl (GelMA)-based bioink. Furthermore, we successfully implanted the 3D printed multi-branched tissue, being connected from the rat carotid artery to the jugular vein direct surgical anastomosis. The work represents a significant step toward in the field of large vascularized tissue fabrication and may have implications for the treatment of organ failure in the future.

Epigenetic Reprogramming via Synergistic Hypomethylation and Hypoxia Enhances the Therapeutic Efficacy of Mesenchymal Stem Cell Extracellular Vesicles for Bone Repair.

Mesenchymal stem cells (MSCs) are a promising cell population for regenerative medicine applications, where paracrine signalling through the extracellular vesicles (EVs) regulates bone tissue homeostasis and development. MSCs are known to reside in low oxygen tension, which promotes osteogenic differentiation via hypoxia-inducible factor-1α activation. Epigenetic reprogramming has emerged as a promising bioengineering strategy to enhance MSC differentiation. Particularly, the process of hypomethylation may enhance osteogenesis through gene activation. Therefore, this study aimed to investigate the synergistic effects of inducing hypomethylation and hypoxia on improving the therapeutic efficacy of EVs derived from human bone marrow MSCs (hBMSCs). The effects of the hypoxia mimetic agent deferoxamine (DFO) and the DNA methyltransferase inhibitor 5-azacytidine (AZT) on hBMSC viability was assessed by quantifying the DNA content. The epigenetic functionality was evaluated by assessing histone acetylation and histone methylation. hBMSC mineralisation was determined by quantifying alkaline phosphate activity, collagen production and calcium deposition. EVs were procured from AZT, DFO or AZT/DFO-treated hBMSCs over a two-week period, with EV size and concentration defined using transmission electron microscopy, nanoflow cytometry and dynamic light scattering. The effects of AZT-EVs, DFO-EVs or AZT/DFO-EVs on the epigenetic functionality and mineralisation of hBMSCs were evaluated. Moreover, the effects of hBMSC-EVs on human umbilical cord vein endothelial cells (HUVECs) angiogenesis was assessed by quantifying pro-angiogenic cytokine release. DFO and AZT caused a time-dose dependent reduction in hBMSC viability. Pre-treatment with AZT, DFO or AZT/DFO augmented the epigenetic functionality of the MSCs through increases in histone acetylation and hypomethylation. AZT, DFO and AZT/DFO pre-treatment significantly enhanced extracellular matrix collagen production and mineralisation in hBMSCs. EVs derived from AZT/DFO-preconditioned hBMSCs (AZT/DFO-EVs) enhanced the hBMSC proliferation, histone acetylation and hypomethylation when compared to EVs derived from AZT-treated, DFO-treated and untreated hBMSCs. Importantly, AZT/DFO-EVs significantly increased osteogenic differentiation and mineralisation of a secondary hBMSC population. Furthermore, AZT/DFO-EVs enhanced the pro-angiogenic cytokine release of HUVECs. Taken together, our findings demonstrate the considerable utility of synergistically inducing hypomethylation and hypoxia to improve the therapeutic efficacy of the MSC-EVs as a cell-free approach for bone regeneration.

Decellularized and Genipin Crosslinked Human Umbilical Cord Artery and Vein for Potential Use as Peripheral Nerve Conduit

Critical gap peripheral nerve injury, commonly caused by motor vehicle accidents, results in dysfunctional nerve and impaired body function. Our study aims to develop a conduit from decellularized and genipin crosslinked human umbilical cord artery and vein for future use in critical nerve gap injury treatments. Human umbilical cord arteries (HUCA) and veins (HUCV) were divided into native (nHUCA and nHUCV), decellularized (dHUCA and dHUCV) and genipin-crosslinked (clHUCA and clHUCV) groups. Both the decellularized and crosslinked groups were decellularized, and subsequently, the clHUCA and clHUCV groups were crosslinked with 0.1%, 0.4% and 0.7% (w/v) genipin. The HUCA and HUCV were then studied for decellularization efficiency, crosslinking index, biodegradation, swelling ratio, ultrastructure analysis, flexibility and mechanical strength. In addition, mesenchymal stem cells isolated from Wharton’s jelly were seeded into HUCA and HUCV for biocompatibility studies. The degradation test showed that nHUCV and dHUCV degraded at day 7 compared to other groups that did not show any degradation even after 21 days. Biocompatibility studies showed that the conduits crosslinked with 0.4% (w/v) genipin were successfully seeded and was having the most amount of seeded cells. In conclusion, the decellularization and genipin crosslinking of human umbilical cord artery and vein enabled successful in fabrication of conduit with suitable properties such as reduced swelling, flexibility, porosity and mechanical strength, with potential in tissue engineering applications.

Vascular and pulmonary effects of ibuprofen on neonatal lung development

BackgroundIbuprofen is a nonsteroidal anti-inflammatory drug that is commonly used to stimulate closure of a patent ductus arteriosus (PDA) in very premature infants and may lead to aberrant neonatal lung development and bronchopulmonary dysplasia (BPD).MethodsWe investigated the effect of ibuprofen on angiogenesis in human umbilical cord vein endothelial cells (HUVECs) and the therapeutic potential of daily treatment with 50 mg/kg of ibuprofen injected subcutaneously in neonatal Wistar rat pups with severe hyperoxia-induced experimental BPD. Parameters investigated included growth, survival, lung histopathology and mRNA expression.ResultsIbuprofen inhibited angiogenesis in HUVECs, as shown by reduced tube formation, migration and cell proliferation via inhibition of the cell cycle S-phase and promotion of apoptosis. Treatment of newborn rat pups with ibuprofen reduced pulmonary vessel density in the developing lung, but also attenuated experimental BPD by reducing lung inflammation, alveolar enlargement, alveolar septum thickness and small arteriolar wall thickening.ConclusionsIn conclusion, ibuprofen has dual effects on lung development: adverse effects on angiogenesis and beneficial effects on alveolarization and inflammation. Therefore, extrapolation of the beneficial effects of ibuprofen to premature infants with BPD should be done with extreme caution.

The isolation of VCAM-1+ endothelial cell-derived extracellular vesicles using microfluidics

Background: Vascular cell adhesion molecule-1 (VCAM-1+) endothelial cell-derived extracellular vesicles (EC-EVs) are augmented in cardiovascular disease, where they can signal the deployment of immune cells from the splenic reserve. Endothelial cells in culture activated with pro-inflammatory tumor necrosis factor-α (TNF-a) also release VCAM-1+ EC-EVs. However, isolating VCAM-1+ EC-EVs from conditioned cell culture media for subsequent in-depth analysis remains challenging. Aim: We utilized the extracellular vesicles (EV) microfluidics herringbone chip (EVHB-Chip), coated with anti-VCAM-1 antibodies, for selective capture of VCAM-1+ cells and EC-EVs. Methods and Results: Engineered EA.hy926 endothelial cells overexpressing VCAM-1 (P < 0.001 versus control) showed increased binding to the VCAM-1- EVHB-Chip versus an IgG device. TNF-α-stimulated human umbilical cord vein endothelial cells (HUVECs) exhibited elevated VCAM-1 protein levels (P < 0.001) and preferential binding to the VCAM-1- EVHB-Chip versus the IgG device. HUVECs stimulated with TNF-α showed differential gene expression of intercellular adhesion molecule-1 (ICAM-1) (P < 0.001) and VCAM-1 (P < 0.001) by digital droplet PCR versus control cells. HUVEC-derived EC-EVs were positive for CD9, CD63, HSP70, and ALIX and had a modal size of 83.5 nm. Control and TNF-α-stimulated HUVEC-derived EC-EV cultures were captured on the VCAM-1- EVHB-Chip, demonstrating selective capture. VCAM-1+ EC-EV were significantly enriched for ICAM-1 (P < 0.001) mRNA transcripts. Conclusion: This study presents a novel approach using the EVHB-Chip, coated with anti-VCAM-1 antibodies and digital droplet PCR for the study of VCAM-1+ EC-EVs. Isolation of VCAM-1+ EC-EV from heterogeneous sources such as conditioned cell culture media holds promise for subsequent detailed characterization, and may facilitate the study of VCAM-1+ EC-EVs in cardiovascular and metabolic diseases, for disease monitoring and therapeutic insights.

In vitro ANGIOGENESIS CAPACITY OF MESENCHYMAL STEM CELLS DERIVED FROM UMBILICAL CORD BLOOD AND ADIPOSE TISSUE

Objective: Angiogenesis plays an essential role in regenerative medicine and damaged tissue repair. Mesenchymal stem cells (MSCs) have been demonstrated to have angiogenic properties and therapeutic potential through their capacity to induce endothelial cell proliferation and tube formation. Besides, bone marrow-derived MSCs have been reported to form a tube-like structure on the Matrigel. In this study, we aimed to check the ability of adipose (AD) and umbilical cord blood (UCB) – derived MSCs in the tube formation and compare them to that of human umbilical cord vein endothelial cells (hUVECs). Methods: The Matrigel was used for the in vitro angiogenesis model. Results: We successfully isolated and cultured ADMSCs, UCBMSCs, and hUVECs with the high expression of specific cell type markers. Moreover, hUVECs could form a tube-like structure network and maintain this morphology for more than 8 hours. ADMSCs and UCBMSCs were able to form the tube-like structure at the early time of 2 hours but failed to retain the structure. Conclusion: ADMSCs and UCBMSCs had potential in the formation of tube-like structures in a short time.

Differential Cytotoxicity of Curcumin-Loaded Micelles on Human Tumor and Stromal Cells.

Although curcumin in the form of nanoparticles has been demonstrated as a potential anti-tumor compound, the impact of curcumin and nanocurcumin in vitro on normal cells and in vivo in animal models is largely unknown. This study evaluated the toxicity of curcumin-loaded micelles in vitro and in vivo on several tumor cell lines, primary stromal cells, and zebrafish embryos. Breast tumor cell line (MCF7) and stromal cells (human umbilical cord vein endothelial cells, human fibroblasts, and human umbilical cord-derived mesenchymal stem cells) were used in this study. A zebrafish embryotoxicity (FET) assay was conducted following the Organisation for Economic Co-operation and Development (OECD) Test 236. Compared to free curcumin, curcumin PM showed higher cytotoxicity to MCF7 cells in both monolayer culture and multicellular tumor spheroids. The curcumin-loaded micelles efficiently penetrated the MCF7 spheroids and induced apoptosis. The nanocurcumin reduced the viability and disturbed the function of stromal cells by suppressing cell migration and tube formation. The micelles demonstrated toxicity to the development of zebrafish embryos. Curcumin-loaded micelles demonstrated toxicity to both tumor and normal primary stromal cells and zebrafish embryos, indicating that the use of nanocurcumin in cancer treatment should be carefully investigated and controlled.

Jatrorrhizine Improves Endothelial Function in Diabetes and Obesity through Suppression of Endoplasmic Reticulum Stress.

Jatrorrhizine (JAT) is one of the major bioactive protoberberine alkaloids found in rhizoma coptidis, which has hypoglycemic and hypolipidemic potential. This study aimed to evaluate the vasoprotective effects of JAT in diabetes and obesity and the underlying mechanism involved. Mouse aortas, carotid arteries and human umbilical cord vein endothelial cells (HUVECs) were treated with risk factors (high glucose or tunicamycin) with and without JAT ex vivo and in vitro. Furthermore, aortas were obtained from mice with chronic treatment: (1) control; (2) diet-induced obese (DIO) mice fed a high-fat diet (45% kcal% fat) for 15 weeks; and (3) DIO mice orally administered JAT at 50 mg/kg/day for the last 5 weeks. High glucose or endoplasmic reticulum (ER) stress inducer tunicamycin impaired acetylcholine-induced endothelium-dependent relaxations (EDRs) in mouse aortas, induced oxidative stress in carotid arteries and HUVECs, downregulated phosphorylations of Akt at Ser473 and eNOS at Ser1177 and enhanced ER stress in mouse aortas and HUVECs, and these impairments were reversed by cotreatment with JAT. JAT increased NO release in high-glucose-treated mouse aortas and HUVECs. In addition, chronic JAT treatment restored endothelial function with EDRs comparable to the control, increased Akt/eNOS phosphorylation, and attenuated ER stress and oxidative stress in aortas from DIO mice. Blood pressure, glucose sensitivity, fatty liver and its morphological change, as well as plasma levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) and plasma lipid profile, were also normalized by JAT treatment. Collectively, our data may be the first to reveal the vasoprotective effect of JAT that ameliorates endothelial dysfunction in diabetes and obesity through enhancement of the Akt/eNOS pathway and NO bioavailability, as well as suppression of ER stress and oxidative stress.

3D engineered neural co-culture model and neurovascular effects of marine fungi-derived citreohybridonol

Marine-based biomolecules are emerging metabolites that have gained attention for developing novel biomaterials, drugs, and pharmaceutical in vitro platforms. Here, we developed a 3D engineered neural co-culture model via a 3D prototyped sliding frame-platform for multi-step UV lithography and investigated the neurovascular potential of citreohybridonol in neuroblastoma treatment. Citreohybridonol was isolated from a sponge-derived fungus Penicillium atrovenetum. The model was characterized by Fourier-transform infrared spectroscopy and scanning electron microscopy analysis. Human umbilical cord vein endothelial cells (HUVECs) and neuroblastoma (SH-SY5Y) cell lines were encapsulated in gelatin methacrylate (GelMA) with and without citreohybridonol. The effect of citreohybridonol on the proliferation capacity of cells was assessed via cell viability and immunostaining assays. GelMA and 3D culture characterization indicated that the cells were successfully encapsulated as axenic and mixed with/without citreohybridonol. The cytotoxic test confirmed that the 3D microenvironment was non-toxic for cultural experiments, and it showed the inhibitory effects of citreohybridonol on SH-SY5Y cells and induced the proliferation of HUVECs. Finally, immunohistochemical staining demonstrated that citreohybridonol suppressed SH-SY5Y cells and induced vascularization of HUVECs in mixed 3D cell culture.

Versatile-in-All-Trades: Multifunctional Boron-Doped Calcium-Deficient Hydroxyapatite Directs Immunomodulation and Regeneration.

Osseointegration of implants depends on several intertwined factors: osteogenesis, angiogenesis, and immunomodulation. Lately, novel reinforcements allowing faster bonding with osseous tissue have been explored intensively. In this study, we hypothesized the use of boron as a major multifunctional ion to confer versatility to calcium-deficient hydroxyapatite (cHA) synthesized by a wet precipitation/microwave reflux method. By synthesis of boron-doped calcium-deficient hydroxyapatite (BcHA), we expected to obtain an osteoimmunomodulatory and regenerative nanoreinforcement. BcHA was found to possess a pure HA phase, a greater surface area (66.41 m2/g, p = 0.028), and cumulative concentrations of Ca (207.87 ± 6.90 mg/mL, p < 0.001) and B (112.70 ± 11.79 mg/mL, p < 0.001) released in comparison to cHA. Osteogenic potential of BcHA was analyzed using human fetal osteoblasts. BcHA resulted in a drastic increase in the ALP activity (1.11 ± 0.11 mmol/gDNA·min, p < 0.001), biomineralization rate, and osteogenic gene expressions compared to cHA. BcHA angiogenic potential was investigated using human umbilical cord vein endothelial cells. Significantly, the highest VEGF-A release (1111.14 ± 87.82 in 4 h, p = 0.009) and angiogenic gene expressions were obtained for BcHA-treated samples. These samples were also observed to induce a more prominent and highly branched tube network. Finally, inflammatory and inflammasome responses toward BcHA were elucidated using human monocyte-derived macrophages differentiated from THP-1s. BcHA exhibited lower CAS-1 release (50.18 ± 5.52 μg/gDNA μg/gDNA) and higher IL-10 release (126.97 ± 15.05 μg/gDNA) than cHA. In addition, BcHA treatment led to increased expression of regenerative genes such as VEGF-A, RANKL, and BMP-2. In vitro results demonstrated that BcHA has tremendous osteogenic, angiogenic, and immunomodulatory potential to be employed as a “versatile-in-all-trades” modality in various bone tissue engineering applications.

Overview of the angiogenic effect of probiotics (Lactobacillus acidophilus and Lactobacillus rhamnosus) at human umbilical vein endothelial cells

Introduction: Angiogenesis (neovascularization), which means new vessel construction, is normal and physiologically, wound healing, embryogenesis, a necessary menstrual cycle it's a mechanism. When taken in appropriate amounts together with or separately with nutrients, mucosal and by regulating systemic immunity, ensuring nutritional and microbial balance in the intestines living nonpatogenic microorganisms that positively affect the health of the host it is called "probiotics". Lactic acid bacteria, the most probiotic microorganisms it constitutes its important group. Where probiotics have an effect on angiogenesis, and it is thought to help heal wounds through the road. With this research indicated that roles of Lactobacillus acidophilus and Lactobacillus rhamnosus on angiogenesis if present to demonstrate in vitro methods and the gene expression responsible for the formation of these effects it is intended to reveal.&#x0D; Material and Method: This study is an experimental study conducted in vitro human umbilical cord vein endothelial cell (HUVEC) MTT test in cell culture with (3-[4,5-Dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide; Thiazolyl blue) evaluation of viability and proliferation wound healing model, tube formation method and gene expression with real rime-polymer chain reaction (RT-PCR) methods of appointment were used.&#x0D; Results: HUVEC cells L. acidophilus 10⁹ CFU\ml after extract application statistical of mRNA expression of VEGF and FGF genes by control group 24 per hour it was found to increase significantly. L. rhamnosus 10 6 CFU\ml and 109 CFU\ml after application of extracts VEGF gene mRNA by control group 24 per hour its expression was found to be statistically significantly increased. Also L. rhamnosus extracts cell proliferation and migration of in vitro wound model it was found to increase statistically significantly.&#x0D; Conclusion: In this study, in vitro L. acidophilus 10⁹ CFU\ml extract and 10⁶ CFU\ml and 10⁹ CFU\ml extract of L. rhamnosus, VEGF gene mRNA revealed to be effective on angiogenesis in HUVEC cells by increasing expression it is.

Protective Role of miR-34c in Hypoxia by Activating Autophagy through BCL2 Repression.

Hypoxia leads to significant cellular stress that has diverse pathological consequences such as cardiovascular diseases and cancers. MicroRNAs (miRNAs) are one of regulators of the adaptive pathway in hypoxia. We identified a hypoxia-induced miRNA, miR-34c, that was significantly upregulated in hypoxic human umbilical cord vein endothelial cells (HUVECs) and in murine blood vessels on day 3 of hindlimb ischemia (HLI). miR-34c directly inhibited BCL2 expression, acting as a toggle switch between apoptosis and autophagy in vitro and in vivo. BCL2 repression by miR-34c activated autophagy, which was evaluated by the expression of LC3-II. Overexpression of miR-34c inhibited apoptosis in HUVEC as well as in a murine model of HLI, and increased cell viability in HUVEC. Importantly, the number of viable cells in the blood vessels following HLI was increased by miR-34c overexpression. Collectively, our findings show that miR-34c plays a protective role in hypoxia, suggesting a novel therapeutic target for hypoxic and ischemic diseases in the blood vessels.