Adhesion of Blood Platelets to the Extracellular Matrix of Cultured Human Endothelial Cells

Sixma, Jan J.; Nievelstein, P F; Zwaginga, Jaap‐Jan; Groot, Philip G. de

doi:10.1111/j.1749-6632.1987.tb33028.x

Cited by 47 publications

(25 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…26 Cells of the second passage were seeded on glass coverslips precoated with gelatin. The ECM was isolated by exposing endothelial cells to 0.1 mol/L NH 4 OH for 15 minutes at room temperature and subsequently washing three times with PBS.…”

Section: Surfacesmentioning

confidence: 99%

“…27 To evaluate aggregate height, perfused Thermanox coverslips were embedded in Epon, and l-/xm-thick cross sections were prepared for evaluation as described. 26 Before and after perfusion, samples of the perfusate were taken to measure single-platelet disappearance (SPD) as a check on platelet clumping caused by handling of blood, as described. 28 For perfusions without dRGDW, SPD values of 20% were found; in the presence of dRGDW, SPD values dropped to 0% because spontaneous platelet clumping was completely prevented.…”

Section: Perfusionsmentioning

confidence: 99%

See 1 more Smart Citation

Aggregate formation is more strongly inhibited at high shear rates by dRGDW, a synthetic RGD-containing peptide.

Saelman¹,

Hese²,

Nieuwenhuis³

et al. 1993

Arterioscler Thromb

View full text Add to dashboard Cite

The effect of D-Arg-Gly-Asp-Trp (dRGDW), a synthetic RGD-containing peptide, on platelet adhesion and aggregate formation on various purified adhesive proteins and the extracellular matrix of endothelial cells was investigated with anticoagulated blood recirculating through a parallel-plate perfusion chamber. Aggregate formation on the extracellular matrix of phorbol myristate acetate (PMA)-stimulated endothelial cells and on collagen type I was more strongly inhibited by dRGDW at higher shear rates than at a low shear rate. Platelet adhesion to the extracellular matrix of nonactivated and PMA-stimulated endothelial cells was inhibited by dRGDW, especially at high shear rates, probably as a consequence of the inhibition of platelet spreading. Inhibition by dRGDW of platelet adhesion to von Willebrand factor, fibronectin, and fibrinogen was almost complete, indicating that platelet adhesion to these substrates is mediated through RGD-directed receptors. Platelet adhesion to laminin was not inhibited by the peptide, whereas platelet adhesion to collagen was increased as a consequence of the inhibition of aggregate formation. Our results show that dRGDW is a strong inhibitor of platelet adhesion and aggregate formation, especially at high shear rates. T he interaction of platelets with components present in the vessel wall is a first and essential step in the hemostatic response. A large number of glycoproteins present in the vessel wall, including von Willebrand factor (vWF), fibronectin, various types of collagen (types I, III, and IV), laminin, thrombospondin, and fibrinogen, are involved in adhesion of platelets, their subsequent spreading, and the induction of aggregate formation.

show abstract

Section: Surfacesmentioning

confidence: 99%

Section: Perfusionsmentioning

confidence: 99%

Aggregate formation is more strongly inhibited at high shear rates by dRGDW, a synthetic RGD-containing peptide.

Saelman¹,

Hese²,

Nieuwenhuis³

et al. 1993

Arterioscler Thromb

View full text Add to dashboard Cite

show abstract

“…Fibrin dep-L osition and platelet aggregation are essential steps in thrombus formation on subendothelial surfaces or perivascular connective tissue. 1 " 3 To what extent fibrin deposition contributes to the formation of the thrombus is influenced by several parameters such as the availability of tissue factor on the vascular surface and local flow characteristics. 4 -5 Fibrin accounts for most of the mass of thrombi found in large veins; such thrombi mainly consist of fibrin and red blood cells.…”

Section: (Arteriosclerosis and Thrombosis 1991;ll:211-220)mentioning

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

View full text Add to dashboard Cite

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

show abstract

“…7 ' 8 To study the vWF dependency in a more natural condition, it was necessary to block the vWF present in the matrix itself. 4 ' 6 Since the FBM lacks endogenous vWF, it is well suited for a study on the vWF dependency on native vessel wall components.…”

mentioning

confidence: 99%

Platelet adhesion to vascular cells. The role of exogenous von Willebrand factor in platelet adhesion.

Nievelstein¹,

Groot²,

d’Alessio³

et al. 1990

Arteriosclerosis

View full text Add to dashboard Cite

Platelet deposition on cultured fibroblasts and on their extracellular matrix (FBM) was Investigated in a flow system with citrated blood and was compared with platelet deposition on cultured endothelial cells, smooth muscle cells, and their extracellular matrices. Platelet deposition was present at all surfaces except on intact endothelial cells. Deposition on FBM consisted of contact platelets, spread platelets, and a few small aggregates. On Intact fibroblasts cells, the surface coverage was lower, and platelets formed aggregates. Factors Involved In primary hemostasls, particularly the wall shear rate, von Willebrand factor (vWF), and fibronectin, were Investigated on FBM. The reactivity of FBM was determined by the passage number of the cultured cells. The vWF was involved In platelet adhesion on FBM at only the high shear rate (>800 8~1). Platelet deposition was independent of plasma fibronectin at all shear rates tested. Matrix-associated fibronectin was Involved in adhesion at low and high wall shear rates. We conclude that FBM can be used as a platelet adhesive surface especially to study the contribution of exogenous vWF to platelet adhesion because FBM does not contain vWF. (Arteriosclerosis 10:462-469, May/June 1990)

show abstract

Adhesion of Blood Platelets to the Extracellular Matrix of Cultured Human Endothelial Cells

Cited by 47 publications

References 25 publications

Aggregate formation is more strongly inhibited at high shear rates by dRGDW, a synthetic RGD-containing peptide.

Aggregate formation is more strongly inhibited at high shear rates by dRGDW, a synthetic RGD-containing peptide.

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

Platelet adhesion to vascular cells. The role of exogenous von Willebrand factor in platelet adhesion.

Contact Info

Product

Resources

About