In-Vitro Model of the Microscale Alveolar Environment

2011 ◽  
Author(s):  
Natalia Higuita-Castro ◽  
Cosmin Mihai ◽  
Derek J. Hansford ◽  
Samir N. Ghadiali
Keyword(s):  
Soft Lithography ◽  
Umbilical Vein ◽  
Three Dimensional ◽  
Multilayered Structure ◽  
Alveolar Epithelial Cells ◽  
Alveolar Epithelial ◽  
Nanoscale System ◽  
Vitro Model ◽  
Umbilical Vein Endothelial Cells

The present work describes the development of a novel micro-nanoscale system that more closely resembles the alveolar-capillary barrier in the lung by recapitulating different parameters of the cellular microenvironment, including fibrous geometry, fiber stiffness, chemistry, and cell-cell interactions. The system consists of a three-dimensional multilayered structure. Two microchannel chambers that resemble alveolar/airway space and the capillary lumen, interfaced with a porous mesh of polymeric nanofibers that act as the basal substrate for seeding lung epithelial and endothelial cell. The top and bottom chambers of the device were fabricated using soft lithography techniques, while the nanofiber mesh was obtained via electrospinning. Human alveolar epithelial cells (A549) and human umbilical vein endothelial cells (HUVEC) were successfully co-cultured using this system. Various cellular and molecular biology techniques are being employed to investigate injury patterns and overall cell responses under different circumstances which mimic various lung disorders such acute lung injury, pulmonary fibrosis and emphysema.

scholarly journals Generation of a Novel In Vitro Model to Study Endothelial Dysfunction from Atherothrombotic Specimens

2021 ◽  
Author(s):  
Susan Gallogly ◽  
Takeshi Fujisawa ◽  
John D. Hung ◽  
Mairi Brittan ◽  
Elizabeth M. Skinner ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Endothelial Dysfunction ◽  
In Vitro Model ◽  
Umbilical Vein ◽  
Coronary Syndrome ◽  
Coronary Endothelial Cells ◽  
Vitro Model ◽  
Umbilical Vein Endothelial Cells

Abstract Purpose Endothelial dysfunction is central to the pathogenesis of acute coronary syndrome. The study of diseased endothelium is very challenging due to inherent difficulties in isolating endothelial cells from the coronary vascular bed. We sought to isolate and characterise coronary endothelial cells from patients undergoing thrombectomy for myocardial infarction to develop a patient-specific in vitro model of endothelial dysfunction. Methods In a prospective cohort study, 49 patients underwent percutaneous coronary intervention with thrombus aspiration. Specimens were cultured, and coronary endothelial outgrowth (CEO) cells were isolated. CEO cells, endothelial cells isolated from peripheral blood, explanted coronary arteries, and umbilical veins were phenotyped and assessed functionally in vitro and in vivo. Results CEO cells were obtained from 27/37 (73%) atherothrombotic specimens and gave rise to cells with cobblestone morphology expressing CD146 (94 ± 6%), CD31 (87 ± 14%), and von Willebrand factor (100 ± 1%). Proliferation of CEO cells was impaired compared to both coronary artery and umbilical vein endothelial cells (population doubling time, 2.5 ± 1.0 versus 1.6 ± 0.3 and 1.2 ± 0.3 days, respectively). Cell migration was also reduced compared to umbilical vein endothelial cells (29 ± 20% versus 85±19%). Importantly, unlike control endothelial cells, dysfunctional CEO cells did not incorporate into new vessels or promote angiogenesis in vivo. Conclusions CEO cells can be reliably isolated and cultured from thrombectomy specimens in patients with acute coronary syndrome. Compared to controls, patient-derived coronary endothelial cells had impaired capacity to proliferate, migrate, and contribute to angiogenesis. CEO cells could be used to identify novel therapeutic targets to enhance endothelial function and prevent acute coronary syndromes.

scholarly journals Effect of aminaphtone on in vitro vascular permeability and capillary-like maintenance

2017 ◽  
Vol 33 (9) ◽  
pp. 592-599 ◽  
Author(s):  
Francesca Felice ◽  
Ester Belardinelli ◽  
Alessandro Frullini ◽  
Tatiana Santoni ◽  
Egidio Imbalzano ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Chronic Venous Insufficiency ◽  
Venous Insufficiency ◽  
Umbilical Vein ◽  
Three Dimensional ◽  
Endothelial Permeability ◽  
Human Umbilical Vein ◽  
Pre Treatment ◽  
Umbilical Vein Endothelial Cells

Objectives Aminaphtone, a naphtohydrochinone used in the treatment of capillary disorders, may affect oedema in chronic venous insufficiency. Aim of study is to investigate the effect of aminaphtone on vascular endothelial permeability in vitro and its effects on three-dimensional capillary-like structures formed by human umbilical vein endothelial cells. Method Human umbilical vein endothelial cells were treated with 50 ng/ml VEGF for 2 h and aminaphtone for 6 h. Permeability assay, VE-cadherin expression and Matrigel assay were performed. Results VEGF-induced permeability was significantly decreased by aminaphtone in a range concentration of 1–20 µg/ml. Aminaphtone restored VE-cadherin expression. Finally, 6 h pre-treatment with aminaphtone significantly preserved capillary-like structures formed by human umbilical vein endothelial cells on Matrigel up to 48 h compared to untreated cells. Conclusions Aminaphtone significantly protects endothelium permeability and stabilises endothelial cells organised in capillary-like structures, modulating VE-cadherin expression. These data might explain the clinical benefit of aminaphtone on chronic venous insufficiency.

scholarly journals Hydrogel-Coated Textile Scaffolds as Three-Dimensional Growth Support for Human Umbilical Vein Endothelial Cells (HUVECs): Possibilities as Coculture System in Liver Tissue Engineering

2002 ◽  
Vol 11 (4) ◽  
pp. 369-377 ◽  
Author(s):  
Makarand V. Risbud ◽  
Erdal Karamuk ◽  
René Moser ◽  
Joerg Mayer
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Three Dimensional ◽  
Mesh Size ◽  
Cell Types ◽  
Cell Number ◽  
Human Umbilical Vein ◽  
Hydrogel Matrix ◽  
Umbilical Vein Endothelial Cells

Three-dimensional (3-D) scaffolds offer an exciting possibility to develop cocultures of various cell types. Here we report chitosan–collagen hydrogel-coated fabric scaffolds with defined mesh size and fiber diameter for 3-D culture of human umbilical vein endothelial cells (HUVECs). These scaffolds did not require pre-coating with fibronectin and they supported proper HUVEC attachment and growth. Scaffolds preserved endothelial cell-specific cobblestone morphology and cells were growing in compartments defined by the textile mesh. HUVECs on the scaffold maintained the property of contact inhibition and did not exhibit overgrowth until the end of in vitro culture (day 6). MTT assay showed that cells had preserved mitochondrial functionality. It was also noted that cell number on the chitosan-coated scaffold was lower than that of collagen-coated scaffolds. Calcein AM and ethidium homodimer (EtD-1) dual staining demonstrated presence of viable and metabolically active cells, indicating growth supportive properties of the scaffolds. Actin labeling revealed absence of actin stress fibers and uniform distribution of F-actin in the cells, indicating their proper attachment to the scaffold matrix. Confocal microscopic studies showed that HUVECs growing on the scaffold had preserved functionality as seen by expression of von Willebrand (vW) factor. Observations also revealed that functional HUVECs were growing at various depths in the hydrogel matrix, thus demonstrating the potential of these scaffolds to support 3-D growth of cells. We foresee the application of this scaffold system in the design of liver bioreactors wherein hepatocytes could be cocultured in parallel with endothelial cells to enhance and preserve liver-specific functions.

scholarly journals Monoclonal antibodies to leukocyte integrins CD11a/CD18 and CD11b/CD18 or intercellular adhesion molecule-1 inhibit chemoattractant-stimulated neutrophil transendothelial migration in vitro

Blood ◽  
1991 ◽  
Vol 78 (8) ◽  
pp. 2089-2097 ◽  
Author(s):  
MB Furie ◽  
MC Tancinco ◽  
CW Smith
Keyword(s):  
Monoclonal Antibodies ◽  
Adhesion Molecule ◽  
Umbilical Vein ◽  
Intercellular Adhesion ◽  
Intercellular Adhesion Molecule ◽  
Intercellular Adhesion Molecule 1 ◽  
Endothelial Monolayers ◽  
Vitro Model ◽  
Umbilical Vein Endothelial Cells

Abstract Intercellular adhesion molecule-1 (ICAM-1) is present on the endothelium and binds to one or more members of the CD11/CD18 family of leukocyte surface integrins. To assess the role of these molecules in mediating chemotaxis of neutrophils across the endothelium, an in vitro model consisting of monolayers of human umbilical vein endothelial cells (HUVEC) grown on amniotic connective tissue was used. Neutrophils placed on the apical sides of these cultures migrated across the endothelium in response to chemoattractants added basally. Monoclonal antibodies (MoAbs) to CD11a, CD11b, and CD18 on the neutrophils inhibited this migration by 52% +/- 11%, 29% +/- 19%, and 90% +/- 7%, respectively. An MoAb to ICAM-1 inhibited transendothelial chemotaxis of the leukocytes by 55% +/- 16%. Inhibition was mediated by binding of the MoAb to ICAM-1 on the HUVEC, rather than by any direct effect of the antibody on the neutrophils. When used in combination, MoAbs to CD11a and to CD11b inhibited migration in a nearly additive fashion. A similar additive effect was observed when MoAbs to CD11b and to ICAM-1 were used together. In contrast, MoAbs to CD11a and to ICAM-1 produced no more inhibition when used in combination than when added singly. These results show that ICAM-1, CD11a/CD18, and CD11b/CD18 all participate in controlling migration of neutrophils across endothelial monolayers in response to chemotactic agents.

scholarly journals An In Vitro Lung System to Assess the Proinflammatory Hazard of Carbon Nanotube Aerosols

2020 ◽  
Vol 21 (15) ◽  
pp. 5335
Author(s):  
Hana Barosova ◽  
Bedia Begum Karakocak ◽  
Dedy Septiadi ◽  
Alke Petri-Fink ◽  
Vicki Stone ◽  
...  
Keyword(s):  
Prolonged Exposure ◽  
Three Dimensional ◽  
Adverse Outcomes ◽  
Alveolar Epithelial Cells ◽  
Proinflammatory Response ◽  
Multiwalled Carbon ◽  
Alveolar Epithelial ◽  
Culture Model ◽  
Air Liquid Interface

In vitro three-dimensional (3D) lung cell models have been thoroughly investigated in recent years and provide a reliable tool to assess the hazard associated with nanomaterials (NMs) released into the air. In this study, a 3D lung co-culture model was optimized to assess the hazard potential of multiwalled carbon nanotubes (MWCNTs), which is known to provoke inflammation and fibrosis, critical adverse outcomes linked to acute and prolonged NM exposure. The lung co-cultures were exposed to MWCNTs at the air-liquid interface (ALI) using the VITROCELL® Cloud system while considering realistic occupational exposure doses. The co-culture model was composed of three human cell lines: alveolar epithelial cells (A549), fibroblasts (MRC-5), and macrophages (differentiated THP-1). The model was exposed to two types of MWCNTs (Mitsui-7 and Nanocyl) at different concentrations (2–10 μg/cm2) to assess the proinflammatory as well as the profibrotic responses after acute (24 h, one exposure) and prolonged (96 h, repeated exposures) exposure cycles. The results showed that acute or prolonged exposure to different concentrations of the tested MWCNTs did not induce cytotoxicity or apparent profibrotic response; however, suggested the onset of proinflammatory response.

scholarly journals Analysis of Low–level Cadmium Exposure Effects on HUVECs (Human Umbilical Vein Endothelial Cells) Cell Viability and Morphology

2019 ◽  
Vol 5 (1) ◽  
pp. 50
Author(s):  
Kristianningrum Dian Sofiana ◽  
Provisia Marthalita Y.W. ◽  
Khotimah Husnul ◽  
M Aris Widodo
Keyword(s):  
Cell Viability ◽  
Cell Morphology ◽  
Umbilical Vein ◽  
Cadmium Exposure ◽  
Control Group ◽  
Correlation Test ◽  
Low Concentration ◽  
Vitro Model ◽  
Umbilical Vein Endothelial Cells

Cadmium is a heavy metal that could be found in daily life. This metal has a toxicity, could contaminate the environment, and affect human health. The main aim of this research was to find the effect of low concentration Cadmium exposure in acute time toward HUVECs cell morphology and viability.In a True experimental research with in vitro model using HUVECs cell, HUVECs cell was divided into four groups. One control group (without CdCl2 induction) and three treatment groups with CdCl2 induction with various concentrations, 0,153 µg/L, 1,53 µg/L and 15,3 µg/L. The trial was repeated five times for each group. Cell morphology was observed with an inverted microscope. Cell viability was examined by MTT assay. Data were analyzed using Kruskal Wallis statistical test and continue with the Man Whitney test. Correlation test was using Spearman.Morphology of treatment group HUVECs cell induced by CdCl2 concentration 15,3 µg/L looked significantly different compared with control group (p<0.05). Cell viability on group HUVECs induced by CdCl2 15,3 µg/L significantly different compared with the control group. The correlation test resulted R= -0,665 with probability 0.001 which means the higher concentration of CdCl2 the lower the viability of cells. Cadmium in low concentration induces cell morphology change and reduce cell viability. Keywords: HUVEC, cadmium, cell morphology, cell viability.  

scholarly journals Monoclonal antibodies to leukocyte integrins CD11a/CD18 and CD11b/CD18 or intercellular adhesion molecule-1 inhibit chemoattractant-stimulated neutrophil transendothelial migration in vitro

Blood ◽  
1991 ◽  
Vol 78 (8) ◽  
pp. 2089-2097 ◽  
Author(s):  
MB Furie ◽  
MC Tancinco ◽  
CW Smith
Keyword(s):  
Monoclonal Antibodies ◽  
Adhesion Molecule ◽  
Umbilical Vein ◽  
Intercellular Adhesion ◽  
Intercellular Adhesion Molecule ◽  
Intercellular Adhesion Molecule 1 ◽  
Endothelial Monolayers ◽  
Vitro Model ◽  
Umbilical Vein Endothelial Cells

Intercellular adhesion molecule-1 (ICAM-1) is present on the endothelium and binds to one or more members of the CD11/CD18 family of leukocyte surface integrins. To assess the role of these molecules in mediating chemotaxis of neutrophils across the endothelium, an in vitro model consisting of monolayers of human umbilical vein endothelial cells (HUVEC) grown on amniotic connective tissue was used. Neutrophils placed on the apical sides of these cultures migrated across the endothelium in response to chemoattractants added basally. Monoclonal antibodies (MoAbs) to CD11a, CD11b, and CD18 on the neutrophils inhibited this migration by 52% +/- 11%, 29% +/- 19%, and 90% +/- 7%, respectively. An MoAb to ICAM-1 inhibited transendothelial chemotaxis of the leukocytes by 55% +/- 16%. Inhibition was mediated by binding of the MoAb to ICAM-1 on the HUVEC, rather than by any direct effect of the antibody on the neutrophils. When used in combination, MoAbs to CD11a and to CD11b inhibited migration in a nearly additive fashion. A similar additive effect was observed when MoAbs to CD11b and to ICAM-1 were used together. In contrast, MoAbs to CD11a and to ICAM-1 produced no more inhibition when used in combination than when added singly. These results show that ICAM-1, CD11a/CD18, and CD11b/CD18 all participate in controlling migration of neutrophils across endothelial monolayers in response to chemotactic agents.

scholarly journals Microcapsules functionalized with neuraminidase can enter vascular endothelial cells in vitro

2014 ◽  
Vol 11 (101) ◽  
pp. 20141027 ◽  
Author(s):  
Weizhi Liu ◽  
Xiaocong Wang ◽  
Ke Bai ◽  
Miao Lin ◽  
Gleb Sukhorukov ◽  
...  
Keyword(s):  
Drug Delivery ◽  
Endothelial Cells ◽  
Mechanical Stability ◽  
Vascular Endothelial Cells ◽  
Umbilical Vein ◽  
Three Dimensional ◽  
Endothelial Barrier ◽  
Live Cells ◽  
Umbilical Vein Endothelial Cells

Microcapsules made of polyelectrolyte multilayers exhibit no or low toxicity, appropriate mechanical stability, variable controllable degradation and can incorporate remote release mechanisms triggered by various stimuli, making them well suited for targeted drug delivery to live cells. This study investigates interactions between microcapsules made of synthetic (i.e. polystyrenesulfonate sodium salt/polyallylamine hydrochloride) or natural (i.e. dextran sulfate/poly- l -arginine) polyelectrolyte and human umbilical vein endothelial cells with particular focus on the effect of the glycocalyx layer on the intake of microcapsules by endothelial cells. Neuraminidase cleaves N -acetyl neuraminic acid residues of glycoproteins and targets the sialic acid component of the glycocalyx on the cell membrane. Three-dimensional confocal images reveal that microcapsules, functionalized with neuraminidase, can be internalized by endothelial cells. Capsules without neuraminidase are blocked by the glycocalyx layer. Uptake of the microcapsules is most significant in the first 2 h. Following their internalization by endothelial cells, biodegradable DS/PArg capsules rupture by day 5; however, there is no obvious change in the shape and integrity of PSS/PAH capsules within the period of observation. Results from the study support our hypothesis that the glycocalyx functions as an endothelial barrier to cross-membrane movement of microcapsules. Neuraminidase-loaded microcapsules can enter endothelial cells by localized cleavage of glycocalyx components with minimum disruption of the glycocalyx layer and therefore have high potential to act as drug delivery vehicles to reach tissues beyond the endothelial barrier of blood vessels.

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