Media conditioned by cultured human vascular endothelial cells inhibit the growth of vascular smooth muscle cells

Willems, C; Astaldi, G; Groot, P. G. De; Janssen, Michiel; Gonsalvez, M.D.; Zeijlemaker, W. P.; Mourik, Jan A. van; Aken, W.G. van

doi:10.1016/0014-4827(82)90332-9

Cited by 148 publications

(45 citation statements)

References 17 publications

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“…Previously, it was known that endothelial cells can regulate VSMC proliferation. 21 In particular, heparin and endothelial cell HSPGs are potent inhibitors of VSMC proliferation and FGF-2 induced mitogenesis. 13,[22][23][24] This regulation is growth state dependent, with subconfluent cultures of endothelial cells stimulating VSMC growth and postconfluent cultures inhibiting VSMC growth.…”

Section: Discussionmentioning

confidence: 99%

Endothelial Cells Provide Feedback Control for Vascular Remodeling Through a Mechanosensitive Autocrine TGF-β Signaling Pathway

Baker

Ettenson

Jonas

et al. 2008

Circulation Research

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Abstract-Mechanical forces are potent modulators of the growth and hypertrophy of vascular cells. We examined the molecular mechanisms through which mechanical force and hypertension modulate endothelial cell regulation of vascular homeostasis. Exposure to mechanical strain increased the paracrine inhibition of vascular smooth muscle cells (VSMCs) by endothelial cells. Mechanical strain stimulated the production of perlecan and heparan sulfate glycosaminoglycans by endothelial cells. By inhibiting the expression of perlecan with an antisense vector we demonstrated that perlecan was essential to the strain-mediated effects on endothelial cell growth control. Mechanical regulation of perlecan expression in endothelial cells was governed by a mechanotransduction pathway requiring autocrine transforming growth factor ␤ (TGF-␤) signaling and intracellular signaling through the ERK pathway.Immunohistochemical staining of the aortae of spontaneously hypertensive rats demonstrated strong correlations between endothelial TGF-␤, phosphorylated signaling intermediates, and arterial thickening. Further, studies on ex vivo arteries exposed to varying levels of pressure demonstrated that ERK and TGF-␤ signaling were required for pressure-induced upregulation of endothelial HSPG. Our findings suggest a novel feedback control mechanism in which net arterial remodeling to hemodynamic forces is controlled by a dynamic interplay between growth stimulatory signals from VSMCs and growth inhibitory signals from endothelial cells. Key Words: mechanical strain Ⅲ hypertension Ⅲ TGF-␤ Ⅲ heparan sulfate proteoglycans Ⅲ perlecan T he endothelium is a dynamic constituent of the vascular system that exerts remarkable control over diverse process such as hemostasis, inflammation, and the regulation of vascular tone. 1,2 In addition to interacting with blood constituents and circulating cells, the vascular endothelium is exposed to a distinct mechanical environment consisting of hemodynamic shear stress and mechanical stretch from blood pressure. As a result, endothelial cells are optimally situated as both sensors and effectors in the vascular remodeling process. An extensive body of scientific work supports the concept that vascular cells respond to their mechanical environment. 3,4 In vitro, shear stress and stretch stimuli regulate endothelial cell production of regulatory molecules including nitric oxide, 5 reactive oxygen species, 6 inflammatory cell adhesion molecules, 7 and extracellular matrix molecules. 8 Similarly, vascular smooth muscle cells (VSMCs) respond to mechanical stimuli by altering their expression of cell adhesion and extracellular matrix molecules 9 as well as increasing proliferation and migration. 10,11 An essential function of the intact endothelium is the paracrine regulation of the growth and phenotype of underlying VSMCs. Soluble heparan sulfate proteoglycans (HSPGs) are intimately involved in endothelial inhibition of VSMC proliferation. 12-14 These complex molecules are composed of a core protein covalently coup...

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Section: Discussionmentioning

confidence: 99%

Endothelial Cells Provide Feedback Control for Vascular Remodeling Through a Mechanosensitive Autocrine TGF-β Signaling Pathway

Baker

Ettenson

Jonas

et al. 2008

Circulation Research

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“…Human vascular endothelial cells were isolated from umbilical cords according to Jaffe et al (18) with some minor modifications (19). The cells were cultured in a medium consisting of RPMI 1640 (Gibco, Paisley, U.K.) and 20% pooled human serum.…”

Section: Methodsmentioning

confidence: 99%

Evidence that the primary binding site of von Willebrand factor that mediates platelet adhesion on subendothelium is not collagen.

Groot¹,

Ottenhof-Rovers²,

Mourik³

et al. 1988

J. Clin. Invest.

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We have studied the binding of von Willebrand factor to extracellular matrices of endothelial cells and to the vessel wall of human umbilical arteries in relation to its function in supporting platelet adhesion. CLB-RAg 201, an MAb against von Willebrand factor, completely inhibits the binding of von Willebrand factor to collagen type I and type III. CLB-RAg 201 does not inhibit the binding of 'SI-von Willebrand factor to extracellular matrices of endothelial cells, to smooth muscle cells, or to the subendothelium. CLB-RAg 201 partly inhibits platelet adhesion to these surfaces, but this directly affects the interaction between von Willebrand factor and platelets and is not due to inhibition of binding of von Willebrand factor to these surfaces.Another MAb, CLB-RAg 38, does not inhibit the binding of von Willebrand factor to collagen. CLB-RAg 38 completely inhibits the binding of von Willebrand factor to extracellular matrices. CLB-RAg 38 inhibits platelet adhesion to cellular matrices completely insofar as it is dependent on plasma von Willebrand factor. CLB-RAg 38 does not inhibit the total binding of von Willebrand factor to subendothelium, as there are too many different binding sites, but it completely inhibits the functional binding sites for von Willebrand factor that support platelet adhesion.The epitopes for CLB-RAg 38 and 201 on the von Willebrand factor molecule are located on different fragments of the molecule. These results indicate that von Willebrand factor binds to subendothelium and matrices of cultured cells by a mechanism that is different from that by which it binds to collagen.

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“…22 The cells were identified by their typical characteristics such as the presence of von Willebrand factor. ECs of the second passage were subcultured on gelatincoated Thermanox coverslips (Flow Laboratories Inc., Rockville, Md.).…”

Section: Cell Culturesmentioning

confidence: 99%

Quantification of fibrin deposition in flowing blood with peroxidase-labeled fibrinogen. High shear rates induce decreased fibrin deposition and appearance of fibrin monomers.

Tijburg¹,

Ijsseldijk²,

Sixma³

et al. 1991

Arterioscler Thromb

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To study fibrin incorporation into thrombi at different wall shear rates, a new method to study fibrin deposition on extracellular matrixes underlying stimulated endothelial cells under flow conditions was developed. For this method, we used fibrinogen labeled with peroxidase (Fg-PO). Fg-PO was fully exchangeable for Fg in the clotting assays tested, and PO activity was bound to fibrin-specific fragments. Fg-PO containing fibrin could be stained for microscopic studies with 3 r 3'-diaminobenzidine and could be quantified by oxidation of phenylenediamine. The absorbance values at 492 nm were converted to fibrin quantities via a standard curve. To study fibrin deposition, Fg-PO was added in trace amounts to whole blood anticoagulated with low-molecularweight heparin, and perfusion studies were performed over endothelial cell matrixes containing tissue factor. In parallel perfusion studies, '"i-labeled Fg was added in trace amounts to whole blood instead of Fg-PO. Both quantitative methods demonstrated decreased fibrin deposition after perfusions at 1,300 sec" 1 compared with fibrin deposition after perfusions at 300 sec"', while fibrinopeptide A generation was independent of the wall shear rate. The decrease in fibrin deposition at 1,300 sec' 1 was accompanied by the appearance of fibrin monomers in the perfusate. This suggested that the decrease in fibrin incorporation at 1,300 sec" 1 was due to the unpaired polymerization of fibrin monomers. This impairment was probably due to a decrease in local fibrin monomer concentration as a result of the increased removal of monomers from the surface at 1,300 sec" Fibrin accounts for most of the mass of thrombi found in large veins; such thrombi mainly consist of fibrin and red blood cells. 6 -7 In major epicardial coronary vessels and arterioles, the amount of fibrin incorporated in the thrombus decreases, but it is still From the Department of Haematology, University Hospital Utrecht, Utrecht, The Netherlands.Supported by the Dutch Heart Foundation (grant 86.044). Address for correspondence: Dr. Pim N.M. Tijburg, Department of Haematology, University Hospital Utrecht, Heidelberglaan 100, PO Box 85500, 3508 GA Utrecht, The Netherlands.Received November 16, 1989; revision accepted October 9, 1990. essential for stabilization of the mixed fibrin/ platelet thrombus. 8 -11 Studies of the fibrin content of thrombi have been notoriously hampered by the lack of methods for its quantification. The presence of fibrin is usually demonstrated by immunofluorescence with the use of radioisotopes or by morphological quantification. 12-15

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Media conditioned by cultured human vascular endothelial cells inhibit the growth of vascular smooth muscle cells

Cited by 148 publications

References 17 publications

Endothelial Cells Provide Feedback Control for Vascular Remodeling Through a Mechanosensitive Autocrine TGF-β Signaling Pathway

Endothelial Cells Provide Feedback Control for Vascular Remodeling Through a Mechanosensitive Autocrine TGF-β Signaling Pathway

Evidence that the primary binding site of von Willebrand factor that mediates platelet adhesion on subendothelium is not collagen.

Quantification of fibrin deposition in flowing blood with peroxidase-labeled fibrinogen. High shear rates induce decreased fibrin deposition and appearance of fibrin monomers.

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