Relationship Between Orientation Angle in Intracellular Stress Fibers and Range of Uniaxial Cyclic Stretch

2000 ◽  
Author(s):  
Hiroshi Yamada ◽  
Tohru Takemasa ◽  
Takami Yamaguchi
Keyword(s):  
Endothelial Cells ◽  
Cultured Cells ◽  
Umbilical Vein ◽  
Orientation Angle ◽  
Mechanical Stimulus ◽  
Cyclic Stretch ◽  
Stress Fibers ◽  
Experimental Result ◽  
Silicone Membrane ◽  
Umbilical Vein Endothelial Cells

Abstract We hypothesized an avoidance of deformation and a limit of sensitivity of cell response to the mechanical stimulus for the orientation of stress fibers in cultured cells on a silicone membrane which was subjected to a uniaxial cyclic stretch. We compared the theoretical prediction with the experimental result of stress fibers in the human umbilical vein endothelial cells under various ranges of uniaxial cyclic stretch (Takemasa et al. 1998). The results showed that the proposed hypothesis predicted the orientation of stress fibers under various ranges of cyclic stretch well.

Involvement of SA channels in orienting response of cultured endothelial cells to cyclic stretch

1998 ◽  
Vol 274 (5) ◽  
pp. H1532-H1538 ◽  
Author(s):  
Keiji Naruse ◽  
Takako Yamada ◽  
Masahiro Sokabe
Keyword(s):  
Endothelial Cells ◽  
Cell Elongation ◽  
Umbilical Vein ◽  
Orienting Response ◽  
Cyclic Stretch ◽  
Silicone Membrane ◽  
Cell Orientation ◽  
Human Umbilical Vein ◽  
Intracellular Ca ◽  
Dependent Cell

The present work was designed to elucidate the involvement of Ca2+-permeable stretch-activated (SA) channels in the orienting response of endothelial cells to uniaxial cyclic stretch. Endothelial cells from human umbilical vein were cultured on an elastic silicone membrane and subjected to uniaxial cyclic stretch (120% in length, 1 Hz). The cells started to change their morphology 15 min after the onset of stretch, and >90% of the cells oriented perpendicularly to the stretch axis after 2 h. Associated with the orienting response, cell elongation proceeded with a slower rate. Both of the orientating and elongating responses were largely inhibited by the removal of external Ca2+ or by Gd3+, a potent blocker for the SA channel, but not by nifedipine. Intracellular Ca2+ concentration ([Ca2+]i) transiently increased in response to uniaxial stretch, and the basal [Ca2+]igradually increased during cyclic stretch. This Ca2+ response was inhibited by the removal of extracellular Ca2+ or by the addition of Gd3+. These results suggest that stretch-dependent Ca2+ influx through SA channels is essential in the stretch-dependent cell orientation and elongation.

Interleukin-13 induces PSGL-1/P–selectin–dependent adhesion of eosinophils, but not neutrophils, to human umbilical vein endothelial cells under flow

Blood ◽  
2000 ◽  
Vol 95 (10) ◽  
pp. 3146-3152 ◽  
Author(s):  
Gerrit Woltmann ◽  
Claire A. McNulty ◽  
Grant Dewson ◽  
Fiona A. Symon ◽  
Andrew J. Wardlaw
Keyword(s):  
Endothelial Cells ◽  
Laser Scanning ◽  
Cultured Cells ◽  
Umbilical Vein ◽  
Allergic Inflammation ◽  
Similar Degree ◽  
Human Umbilical Vein ◽  
Laser Scanning Cytometry ◽  
Umbilical Vein Endothelial Cells ◽  
Necrosis Factor

Selective eosinophil accumulation is a hallmark of diseases such as asthma. In a model of chronic eosinophilic inflammation, we have previously shown that the tethering step in eosinophil adhesion is mediated by PSGL-1 binding to P-selectin. The Th2-associated cytokine IL-13 is of potential importance in allergic disease. We have therefore investigated whether IL-13 can mediate eosinophil binding to human umbilical vein endothelial cells (HUVEC) through P-selectin. IL-13 caused dose- and time-dependent increases of P-selectin expression, as assessed by flow and laser scanning cytometry. A similar degree of expression was observed with IL-4. There was no effect on E-selectin or ICAM-1 expression. Tumor necrosis factor- induced the expression of VCAM-1, E-selectin, and ICAM-1 but had no effect on P-selectin expression. IL-13 increased the production of mRNA for surface and soluble variants of P-selectin. Under flow conditions, eosinophils, but not neutrophils, showed enhanced binding to IL-13 and to IL-4–stimulated HUVEC compared to medium-cultured cells. Eosinophil adhesion was completely inhibited by a blocking monoclonal antibody against PSGL-1 and P-selectin. Anti–VLA-4 and anti–VCAM-1 antibodies inhibited binding to a lesser extent. Thus, at physiologic levels of expression induced by Th2 cytokines, P-selectin/PSGL-1 supported eosinophil but not neutrophil adhesion. This mechanism is likely to be a key event leading to the selective accumulation of eosinophils in allergic inflammation.

The Influence of Cyclic and Uniform Shear Stresses Concurrent with Cyclic Stretch on the Gene Expression of Human Umbilical Vein Endothelial Cells

2013 ◽  
Vol 3 (6) ◽  
pp. 673-678 ◽  
Author(s):  
Shahrokh Shojaei ◽  
Mohammad Tafazzoli-Shadpour ◽  
Mohammad Ali Shokrgozar ◽  
Nooshin Haghighipour ◽  
Najmeh Safaei ◽  
...  
Keyword(s):  
Gene Expression ◽  
Endothelial Cells ◽  
Umbilical Vein ◽  
Cyclic Stretch ◽  
Shear Stresses ◽  
Human Umbilical Vein ◽  
Uniform Shear ◽  
Umbilical Vein Endothelial Cells

Dynamics of integrin clustering at focal contacts of endothelial cells studied by multimode imaging microscopy

2001 ◽  
Vol 114 (17) ◽  
pp. 3125-3135 ◽  
Author(s):  
Keisuke Kawakami ◽  
Hitoshi Tatsumi ◽  
Masahiro Sokabe
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Time Lapse ◽  
Stress Fibers ◽  
Intracellular Space ◽  
Focal Contacts ◽  
Actin Fiber ◽  
Glass Cover Slip ◽  
Integrin Clustering ◽  
Umbilical Vein Endothelial Cells

Human umbilical vein endothelial cells were stained with FITC-labeled anti-β1 integrin antibody and plated on a glass cover slip to elucidate the mechanism of integrin clustering during focal contact formation. The process of integrin clustering was observed by time-lapse total-internal-reflection fluorescence microscopy, which can selectively visualize the labeled integrins at the basal surface of living cells. The clustering of integrins at focal contacts started at 1 hour after plating and individual clusters kept growing for ∼6 hours. Most integrin clusters (∼80%) elongated towards the cell center or along the cell margin at a rate of 0.29±0.24 μm minute−1. Photobleaching and recovery experiments with evanescent illumination revealed that the integrins at the extending tip of the clusters were supplied from the intracellular space. Simultaneous time-lapse imaging of exocytosis of integrin-containing vesicles and elongating focal contacts showed that most exocytosis occurred at or near the focal contacts followed by their elongation. Double staining of F-actins and integrins demonstrated that stress fibers were located near the integrin clusters and that intracellular punctate integrins were associated with these stress fibers. These results suggest that the clustering of integrins is mediated by actin-fiber-dependent translocation of integrins to the extending tip of focal contacts.

Interleukin-13 induces PSGL-1/P–selectin–dependent adhesion of eosinophils, but not neutrophils, to human umbilical vein endothelial cells under flow

Blood ◽  
2000 ◽  
Vol 95 (10) ◽  
pp. 3146-3152 ◽  
Author(s):  
Gerrit Woltmann ◽  
Claire A. McNulty ◽  
Grant Dewson ◽  
Fiona A. Symon ◽  
Andrew J. Wardlaw
Keyword(s):  
Endothelial Cells ◽  
Laser Scanning ◽  
Cultured Cells ◽  
Umbilical Vein ◽  
Allergic Inflammation ◽  
Similar Degree ◽  
Human Umbilical Vein ◽  
Laser Scanning Cytometry ◽  
Umbilical Vein Endothelial Cells ◽  
Necrosis Factor

Abstract Selective eosinophil accumulation is a hallmark of diseases such as asthma. In a model of chronic eosinophilic inflammation, we have previously shown that the tethering step in eosinophil adhesion is mediated by PSGL-1 binding to P-selectin. The Th2-associated cytokine IL-13 is of potential importance in allergic disease. We have therefore investigated whether IL-13 can mediate eosinophil binding to human umbilical vein endothelial cells (HUVEC) through P-selectin. IL-13 caused dose- and time-dependent increases of P-selectin expression, as assessed by flow and laser scanning cytometry. A similar degree of expression was observed with IL-4. There was no effect on E-selectin or ICAM-1 expression. Tumor necrosis factor- induced the expression of VCAM-1, E-selectin, and ICAM-1 but had no effect on P-selectin expression. IL-13 increased the production of mRNA for surface and soluble variants of P-selectin. Under flow conditions, eosinophils, but not neutrophils, showed enhanced binding to IL-13 and to IL-4–stimulated HUVEC compared to medium-cultured cells. Eosinophil adhesion was completely inhibited by a blocking monoclonal antibody against PSGL-1 and P-selectin. Anti–VLA-4 and anti–VCAM-1 antibodies inhibited binding to a lesser extent. Thus, at physiologic levels of expression induced by Th2 cytokines, P-selectin/PSGL-1 supported eosinophil but not neutrophil adhesion. This mechanism is likely to be a key event leading to the selective accumulation of eosinophils in allergic inflammation.

scholarly journals Mechano-chemical control of human endothelium orientation and size.

1989 ◽  
Vol 109 (1) ◽  
pp. 331-339 ◽  
Author(s):  
V P Shirinsky ◽  
A S Antonov ◽  
K G Birukov ◽  
A V Sobolevsky ◽  
Y A Romanov ◽  
...  
Keyword(s):  
Adenylate Cyclase ◽  
Umbilical Vein ◽  
Giant Cells ◽  
Stress Fiber ◽  
Fiber Formation ◽  
Cyclic Stretch ◽  
Stress Fibers ◽  
Conventional Culture ◽  
Orientation Response ◽  
Umbilical Vein Endothelial Cells

Human umbilical vein endothelial cells (EC) were grown on elastic silicone membranes subjected to cyclic stretch, simulating arterial wall motion. Stretching conditions (20% amplitude, 52 cycle/min) stimulated stress fiber formation and their orientation transversely to the strain direction. Cell bodies aligned along the same axis after the actin cytoskeleton. EC orientation response was inhibited by the adenylate cyclase activator, forskolin (10(-5) M), which caused stress fiber disassembly and the redistribution of F-actin to the cortical cytoplasm. Preoriented EC depleted of stress fibers by forskolin treatment retained their aligned state. Thus, stress fibers are essential for the process of EC orientation induced by repeated strain, but not for the maintenance of EC orientation. The monolayer formed by EC grown to confluence in conditions of intermittent strain consisted of uniform elongated cells and was resistant to deformation. In contrast, the monolayer assembled in stationary conditions was less compliant and exposed local denudations on initiation of stretching. When stretched in the presence of 10(-5) M forskolin it rapidly (3-4 h) reestablished integrity but gained a heterogeneous appearance since denuded areas were covered by giant cells. The protective effect of forskolin was because of the stimulation of EC spreading. This feature of forskolin was demonstrated while studying its action on EC spreading and repair of a scratched EC monolayer in conventional culture. Thus mechanical deformation and adenylate cyclase activity may be important factors in the control of endothelium morphology in human arteries.

scholarly journals P2Y2 receptor modulates shear stress-induced cell alignment and actin stress fibers in human umbilical vein endothelial cells

2016 ◽  
Vol 74 (4) ◽  
pp. 731-746 ◽  
Author(s):  
Ramasri Sathanoori ◽  
Paulina Bryl-Gorecka ◽  
Christa E. Müller ◽  
Laurie Erb ◽  
Gary A. Weisman ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Umbilical Vein ◽  
Stress Fibers ◽  
Cell Alignment ◽  
P2y2 Receptor ◽  
Human Umbilical Vein ◽  
Umbilical Vein Endothelial Cells ◽  
Actin Stress Fibers

scholarly journals Cytokine-Mediated Activation of Cultured CNS Microvessels: A System for Examining Antigenic Modulation of CNS Endothelial Cells, and Evidence for Long-Term Expression of the Adhesion Protein E-Selectin

1994 ◽  
Vol 14 (5) ◽  
pp. 837-844 ◽  
Author(s):  
Paula Dore-Duffy ◽  
Ruth A. Washington ◽  
Roumen Balabanov
Keyword(s):  
Endothelial Cells ◽  
Cultured Cells ◽  
Umbilical Vein ◽  
Necrosis Factor Alpha ◽  
Antigenic Modulation ◽  
Adhesion Protein ◽  
Activation Antigens ◽  
Umbilical Vein Endothelial Cells ◽  
Cell Adhesion Molecule 1

Much of what is known of endothelial responses to cytokines has been derived from in vitro studies using cultured human umbilical vein endothelial cells (EC). Less is known of CNS EC responses and whether intact endothelium responds similarly to cultured cells. We have used techniques by which rat CNS microvessels can be isolated, then cultured in vitro, to study the response of intact endothelium to activation with cytokines. These microvessels are composed of viable EC and perivascular cells, predominantly pericytes. Expression of EC activation antigens in multicellular systems such as cultured microvessels can be assessed quantitatively using immunofluorescence laser cytometry. Interferon gamma increased immunologically reactive major histocompatibility complex class II antigens (<300 to 2,398 ± 225 average fluorescence intensity), while tumor necrosis factor alpha induced an increase in vascular cell adhesion molecule-1 (2,167 ± 171) and E-selectin (1,628 ± 315). CNS EC appeared to respond similarly to cultured EC with the exception that E-selectin expression was not transiently expressed but was maintained by microvessel EC for 24 and 48 h. Cultured CNS microvessels provide a good system for studying EC activation.

The Effect of Diameter of Electronspun PGA Scaffold for Biological Behaviour of Human Umbilical Vein Endothelial Cells

2007 ◽  
Vol 342-343 ◽  
pp. 237-240 ◽  
Author(s):  
Furong Tian ◽  
Hossein Hossinkhani ◽  
Yoshiro Yokoyama ◽  
Giovani Gomes Estrada ◽  
Hisatoshi Kobayashi
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Glycolic Acid ◽  
Vascular Tissue ◽  
Umbilical Vein ◽  
Silicone Membrane ◽  
Biological Behaviour ◽  
Human Umbilical Vein ◽  
Scaffold Materials ◽  
Umbilical Vein Endothelial Cells

The objective of this work was to investigate cell adhesion, Poly Glycolic Acid (PGA) and PGA/ collagen nano-fibers on the silicone membrane. PGA with the weight-mixing ration of 40% was fabricated through the electronspun technique. The behaviors of Human Umbilical Vein Endothelial cells on these scaffolds are evaluated. The highest cell adhesion was observed in the PGA/collagen fibers with the diameter of 500 nm. This study indicates the effect of nano-fibers on the Human Umbilical Vein Endothelial cells for better understanding of interactions of cells with scaffold materials. Such information will have important implications for implantable vascular tissue engineering constructs.

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