Models Of Granulocyte-Mediated Endothelial Injury In Vitro

1981 ◽  
Author(s):  
G Vercellotti ◽  
P Flynn ◽  
D Weisdorf ◽  
C J Lammi-Keefe ◽  
H Jacob ◽  
...  
Keyword(s):  
Lipid A ◽  
Umbilical Vein ◽  
Endothelial Damage ◽  
Endothelial Injury ◽  
Free Radical Scavengers ◽  
Methyl Prednisolone ◽  
Vitro Model ◽  
Umbilical Vein Endothelial Cells

To determine the role of neutrophil (PMN) induced vascular injury during inflammation an in vitro model of endothelial damage was investigated. Injury to human umbilical vein endothelial cells (EC) labeled with 51Cr or 14c sodium arachidonate was monitored by specific release of these labels or their products. Several agents were capable of triggering PMN to induce significant EC injury: these include activated serum complement (C') opsonized particles, serotonin , phorbol myristate acetate, and the lipid A moiety of endotoxin. PMN must adhere closely to the EC for effective cytotoxicity, since agents which retard PMN adherence (cytochalasin B, methyl prednisolone) inhibit 51Cr release.Lyscsorcal proteases did not mediate PMN induced endothelial injury since there was no correlation between release and injury, and soluble stimuli which did not release lysosomal contents induced injury. Free radical scavengers such as SOD/catalase, and a-tocopherol significantly reduced PMN mediated endothelial injury implying that PMN generated reactive oxygen species were responsible for this damage.To further study PMN mediated endothelial injury, other inflammatory agents were also utilized. C' activated PMN’s exposed to Ibuprofen (I) (50 μg/ml) but not Aspirin (ASA) (200 μg/ml) induced no EC injury (51cr releasey Furthermore, it was shown that I inhibits O- 2 production, blocks release of PMN lysozyme and glucuronidase, and inhibits aggregation of C' stimulated PMN’s. ASA at doses of 500 μg/ml (clinically toxic), failed to inhibit these in vitro activities. This data suggests that I’s anti-inflammatory affect may be expressed through inhibition of PMN functions. Since I and ASA both inhibit cyclooxygenase, but only I modulated PMN induced endothelial injury, these agents may provide useful probes to elucidate PMN-endothelial interactions in vivo.

Protective Effect of Vitamin E on Immune Triggered, Granulocyte Mediated Endothelial Injury

1984 ◽  
Vol 51 (01) ◽  
pp. 089-092 ◽  
Author(s):  
M A Boogaerts ◽  
J Van de Broeck ◽  
H Deckmyn ◽  
C Roelant ◽  
J Vermylen ◽  
...  
Keyword(s):  
Vitamin E ◽  
Protective Effect ◽  
Umbilical Vein ◽  
Endothelial Damage ◽  
Endothelial Injury ◽  
Mediated Effects ◽  
Anti Oxidant ◽  
Endothelial Cell Cultures ◽  
Vitro Model

SummaryThe effect of alfa-tocopherol on the cell-cell interactions at the vessel wall were studied, using an in vitro model of human umbilical vein endothelial cell cultures (HUEC). Immune triggered granulocytes (PMN) will adhere to and damage HUEC and platelets enhance this PMN mediated endothelial injury. When HUEC are cultured in the presence of vitamin E, 51Cr-leakage induced by complement stimulated PMN is significantly decreased and the enhanced cytotoxicity by platelets is completely abolished (p <0.001).The inhibition of PMN induced endothelial injury is directly correlated to a diminished adherence of PMN to vitamin E- cultured HUEC (p <0.001), which may be mediated by an increase of both basal and stimulated endogenous prostacyclin (PGI2) from alfa-tocopherol-treated HUEC (p <0.025). The vitamin E-effect is abolished by incubation of HUEC with the irreversible cyclo-oxygenase inhibitor, acetylsalicylic acid, but the addition of exogenous PGI2 could not reproduce the vitamin E-mediated effects.We conclude that vitamin E exerts a protective effect on immune triggered endothelial damage, partly by increasing the endogenous anti-oxidant potential, partly by modulating intrinsic endothelial prostaglandin production. The failure to reproduce vitamin E-protection by exogenously added PGI2 may suggest additional, not yet elucidated vitamin E-effects on endothelial metabolism.

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.

Structural analysis of endothelial cell mediated prolongation of t-PA driven lysis of an “aged clot”

1995 ◽  
Vol 53 ◽  
pp. 798-799
Author(s):  
W.G. Jerome ◽  
S. Handt ◽  
R.R. Hantgan
Keyword(s):  
Heart Attack ◽  
Umbilical Vein ◽  
Controlled Study ◽  
Voltage Electron ◽  
Tnf Α ◽  
Glass Tubes ◽  
Vitro Model ◽  
Umbilical Vein Endothelial Cells

In an effort to reestablish blood flow in blocked coronary arteries, heart attack victims are now routinely treated with thrombolytic (clot busting) drugs. However, thrombolytic therapy fails to restore sufficient flow in approximately 20% of cases. The reasons for these failures are undoubtedly multifactorial and difficult to study in vivo. We have developed an “in vitro” model blood vessel which allows controlled study of factors influencing thrombolysis. In the studies reported here, we investigated the 3-D distribution of HUVEC secreted PAI-1 within the clot using immuno-gold cytochemistry and intermediate voltage electron microscopy (IVEM).Human umbilical vein endothelial cells (HUVEC) were grown on the inside surface of fibronectin-coated glass tubes. The cells are stimulated for 4 hours with 1000 U/ml of TNF-α. Clot formation was initiated by addition of 2.3 μM fibrinogen, 100 nM plasminogen, and then 2 nM thrombin in HEPES buffered Gey's salts + 0.5% human serum albumin. After aging the clot for up to 4 hours (to mimic the time delay between heart attack and treatment), 0.5 nM rt-PA (thrombolytic agent) was perfused through the clot for 10 min. at 5 ml/hr. Formation and dissolution of the clot were monitored by laser light scattering.

Enkephalins Modify Granulocyte-Endothelial Interactions by Stimulating Prostacyclin Production

1983 ◽  
Vol 50 (02) ◽  
pp. 572-575 ◽  
Author(s):  
M A Boogaerts ◽  
J Vermylen ◽  
H Deckmyn ◽  
C Roelant ◽  
R L Verwilghen ◽  
...  
Keyword(s):  
Umbilical Vein ◽  
Vascular Diseases ◽  
Endothelial Damage ◽  
Base Line ◽  
Endogenous Opioid Peptides ◽  
Endogenous Opioid ◽  
Umbilical Vein Endothelial Cells ◽  
Prostacyclin Production

SummaryGranulocyte (PMN)-endothehal interactions have been implicated in the primary events of vascular injury and atherogenesis. We now present data which show that endogenous opioid peptides, e.g. enkephalins (ENK), dampen immune-triggered, granulocyte-induced endothelial damage by enhancing prostacyclin production. Concurrent exposure of human umbilical vein endothelial cells (HUEC) to Met55-enkephalin increased arachidonic acid (AA, 20 uM) and thrombin (T, 10 U/ml) induced 6-keto-PGF1α-release to respectively 197.2 ± 28.1% and 204.1 ± 17.8% (mean ± SEM) of base line stimulation (p < 0.025). The increases noted were significant at ENK-concentrations as low as 10-12M. Simultaneous addition of naloxone with ENK completely abolished the enhanced 6-keto-PGF1α- release. Addition of a protease resistant enkephalin analogue significantly (p <0.01 over several different concentrations) reduced PMN adherence to HUEC; concomitantly 51Cr-leakage from HUEC that had been exposed to PMN plus activated serum complement was decreased. The even further enhanced 51Cr- leakage that occurs when platelet release products (e.g. serotonin) are included is also decreased by added enkephalin. These data suggest that endogenous neurotransmitters may affect endothelial prostaglandin metabolism, and by so doing provide a protective effect during in vitro, and perhaps in vivo, PMN mediated endothelial injury. This link between neurohumoral and inflammatory systems might enhance our understanding of stress- related phenomena in inflammation and vascular diseases.

Ultrastructural observations on the in vitro cytoadherence of Plasmodium falciparum infected red blood cells

1990 ◽  
Vol 48 (3) ◽  
pp. 914-915
Author(s):  
D.J.P. Ferguson ◽  
A.R. Berendt ◽  
J. Tansey ◽  
K. Marsh ◽  
C.I. Newbold
Keyword(s):  
Plasmodium Falciparum ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Umbilical Vein ◽  
Cell Types ◽  
Amelanotic Melanoma ◽  
Cytoplasmic Process ◽  
Umbilical Vein Endothelial Cells

In human malaria, the most serious clinical manifestation is cerebral malaria (CM) due to infection with Plasmodium falciparum. The pathology of CM is thought to relate to the fact that red blood cells containing mature forms of the parasite (PRBC) cytoadhere or sequester to post capillary venules of various tissues including the brain. This in vivo phenomenon has been studied in vitro by examining the cytoadherence of PRBCs to various cell types and purified proteins. To date, three Ijiost receptor molecules have been identified; CD36, ICAM-1 and thrombospondin. The specific changes in the PRBC membrane which mediate cytoadherence are less well understood, but they include the sub-membranous deposition of electron-dense material resulting in surface deformations called knobs. Knobs were thought to be essential for cytoadherence, lput recent work has shown that certain knob-negative (K-) lines can cytoadhere. In the present study, we have used electron microscopy to re-examine the interactions between K+ PRBCs and both C32 amelanotic melanoma cells and human umbilical vein endothelial cells (HUVEC).We confirm previous data demonstrating that C32 cells possess numerous microvilli which adhere to the PRBC, mainly via the knobs (Fig. 1). In contrast, the HUVEC were relatively smooth and the PRBCs appeared partially flattened onto the cell surface (Fig. 2). Furthermore, many of the PRBCs exhibited an invagination of the limiting membrane in the attachment zone, often containing a cytoplasmic process from the endothelial cell (Fig. 2).

scholarly journals SiRNA Directed Against Annexin II Receptor Inhibits Angiogenesis via Suppressing MMP2 and MMP9 Expression

2015 ◽  
Vol 35 (3) ◽  
pp. 875-884 ◽  
Author(s):  
Hongyuan Song ◽  
Dongyan Pan ◽  
Weifeng Sun ◽  
Cao Gu ◽  
Yuelu Zhang ◽  
...  
Keyword(s):  
Matrix Metalloproteinase ◽  
Umbilical Vein ◽  
Tube Formation ◽  
Annexin Ii ◽  
Mmp9 Expression ◽  
Cell Arrest ◽  
Umbilical Vein Endothelial Cells

Background/Aims: Annexin II receptor (AXIIR) is able to mediate Annexin II signal and induce apoptosis, but its role in angiogenesis remains unclear. This study tries to investigate the role of AXIIR in angiogenesis and the plausible molecular mechanism. Methods/Results: RNA interference technology was used to silence AXIIR, and the subsequent effects in vitro and in vivo were evaluated thereafter. Our data indicated that human umbilical vein endothelial cells (HUVECs) expressed AXIIR and knockdown of AXIIR significantly inhibited HUVECs proliferation, adhesion, migration, and tube formation in vitro and suppressed angiogenesis in vivo. Furthermore, AXIIR siRNA induced cell arrest in the S/G2 phase while had no effect on cell apoptosis. We found that these subsequent effects might be via suppressing the expression of matrix metalloproteinase 2and matrix metalloproteinase 9. Conclusion: AXIIR participates in angiogenesis, and may be a potential therapeutic target for angiogenesis related diseases.

scholarly journals Topical Application of Hyaluronic Acid-RGD Peptide-Coated Gelatin/Epigallocatechin-3 Gallate (EGCG) Nanoparticles Inhibits Corneal Neovascularization via Inhibition of VEGF Production

Pharmaceutics ◽  
2020 ◽  
Vol 12 (5) ◽  
pp. 404 ◽  
Author(s):  
Takuya Miyagawa ◽  
Zhi-Yu Chen ◽  
Che-Yi Chang ◽  
Ko-Hua Chen ◽  
Yang-Kao Wang ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Topical Application ◽  
Umbilical Vein ◽  
Corneal Neovascularization ◽  
Tube Formation ◽  
Antiangiogenic Effect ◽  
Arginine Glycine Aspartic Acid ◽  
Umbilical Vein Endothelial Cells

Neovascularization (NV) of the cornea disrupts vision which leads to blindness. Investigation of antiangiogenic, slow-release and biocompatible approaches for treating corneal NV is of great importance. We designed an eye drop formulation containing gelatin/epigallocatechin-3-gallate (EGCG) nanoparticles (NPs) for targeted therapy in corneal NV. Gelatin-EGCG self-assembled NPs with hyaluronic acid (HA) coating on its surface (named GEH) and hyaluronic acid conjugated with arginine-glycine-aspartic acid (RGD) (GEH-RGD) were synthesized. Human umbilical vein endothelial cells (HUVECs) were used to evaluate the antiangiogenic effect of GEH-RGD NPs in vitro. Moreover, a mouse model of chemical corneal cauterization was employed to evaluate the antiangiogenic effects of GEH-RGD NPs in vivo. GEH-RGD NP treatment significantly reduced endothelial cell tube formation and inhibited metalloproteinase (MMP)-2 and MMP-9 activity in HUVECs in vitro. Topical application of GEH-RGD NPs (once daily for a week) significantly attenuated the formation of pathological vessels in the mouse cornea after chemical cauterization. Reduction in both vascular endothelial growth factor (VEGF) and MMP-9 protein in the GEH-RGD NP-treated cauterized corneas was observed. These results confirm the molecular mechanism of the antiangiogenic effect of GEH-RGD NPs in suppressing pathological corneal NV.

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 Apigenin Suppresses Angiogenesis by Inhibiting Tube Formation and Inducing Apoptosis

2016 ◽  
Vol 11 (10) ◽  
pp. 1934578X1601101
Author(s):  
Hyun Ju Kim ◽  
Mok-Ryeon Ahn
Keyword(s):  
Umbilical Vein ◽  
Chorioallantoic Membrane ◽  
Tube Formation ◽  
Anticancer Activities ◽  
Apoptotic Pathway ◽  
Inhibition Of Angiogenesis ◽  
Induced Apoptosis ◽  
Umbilical Vein Endothelial Cells

Apigenin has been reported to exert angiogenic and anticancer activities in vitro. The mechanism of inhibition of angiogenesis by apigenin, however, has not been well-established. In this study, we investigated whether apigenin not only inhibited tube formation but also induced apoptosis in human umbilical vein endothelial cells (HUVECs). Furthermore, strong antiangiogenic activity of apigenin was observed in the in vivo assay using chick embryo chorioallantoic membrane (CAM). We also analyzed changes in survival signals and the apoptotic pathway through Western blotting. The results indicate that apigenin exerts its antiangiogenic effects through induction of endothelial apoptosis.

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