Comparison of Tissue Factor Pathway in Human Umbilical Vein and Adult Saphenous Vein Endothelial Cells: Implications for Newborn Hemostasis and for Laboratory Models of Endothelial Cell Function

Grabowski, Eríc F.; Carter, Christine A.; Ingelfinger, Julie R.; Tsukurov, Olga; Conroy, Nancy; Abbott, William M.; Orkin, Roslyn W.

doi:10.1203/00006450-199912000-00001

Cited by 149 publications

(113 citation statements)

References 23 publications

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“…13,14 Therefore, cell surfaceassociated TFPI need not increase markedly with shear stress (see below). We also have previously reported no significant change in TFPI secretion after cytokine activation, 15 an observation consistent with that of Ameri et al, 18 who found increases in TFPI levels only of the order of 25%.…”

Section: Discussionsupporting

confidence: 77%

“…In contrast, the reduction in functional TF of the present study was observed at 0.68 dyne/cm 2 in a single-pass system, which eliminates the possibility of such TFPI accumulation. This is not a trivial matter, because the presence of an antibody directed against Kunitz domain I of human TFPI was found in the present work and in previous work 15 to more than double measurable production of factor Xa under static conditions, indicating a marked downregulation by TFPI of EC functional TF even under static circumstances. We hypothesize that the downregulation of functional TF under flow conditions is due chiefly to TFPI when changes in shear stress are acute (eg, nearly simultaneous with the onset of cytokine exposure) but may be due as well to downregulation of TF mRNA (and possible upregulation of TFPI mRNA) when changes in shear stress are more chronic (eg, lasting several hours or more).…”

Section: Discussionmentioning

confidence: 53%

“…In view of the reported equilibrium 13,14 that is believed to exist between supernatant "free" TFPI and EC-associated TFPI, we previously 15 characterized the affinity in solution of a monoclonal antibody directed against Kunitz domain 1 of human TFPI (American Diagnostica) for EC-associated TFPI in terms of MichaelisMenten kinetics. For human umbilical vein ECs, 15 we estimated an apparent rate constant, k, of 8.68Ϯ3.08 g/mL (meanϮSE) and a maximum production rate of factor Xa, F max , of 1.385Ϯ0.191 fmol/cm 2 per 100 000 cells (meanϮSE).…”

Section: Anti-tfpi Assaymentioning

confidence: 99%

“…For human umbilical vein ECs, 15 we estimated an apparent rate constant, k, of 8.68Ϯ3.08 g/mL (meanϮSE) and a maximum production rate of factor Xa, F max , of 1.385Ϯ0.191 fmol/cm 2 per 100 000 cells (meanϮSE). Therefore, some monolayers were washed with HEPES buffer with 0.1% bovine serum albumin and incubated with 100 nmol/L anti-TFPI for 30 minutes at 37°C.…”

Section: Anti-tfpi Assaymentioning

confidence: 99%

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Shear Stress Decreases Endothelial Cell Tissue Factor Activity by Augmenting Secretion of Tissue Factor Pathway Inhibitor

Grabowski

Reininger

Petteruti

et al. 2001

ATVB

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Abstract-Monolayers of human umbilical vein endothelial cells were activated with 50 U/mL interleukin-1␣ (IL-1␣) for 3 hours and simultaneously conditioned with shear stresses of 0, 0.68, or 13.2 dyne/cm 2 in a parallel-plate flow chamber. In the presence of an inflow buffer containing 100 nmol/L factor X and 10 nmol/L factor VII, production of factor Xa, a measure of functional tissue factor (TF), was determined as the product of outflow concentration of factor Xa (chromogenic assay performed under quasi-static flow conditions after the shear period) and flow rate. Similarly, production of TF pathway inhibitor (TFPI) was estimated as the product of antigenic TFPI (by enzyme-linked immunosorbent assay) in the supernatant and flow rate. In parallel experiments, total RNA was isolated for determination of amplification products of TF mRNA by reverse transcription-polymerase chain reaction. We found that shear stress reduced factor Xa production (meanϮSE; nϭnumber of experiments) from 13.33Ϯ1.14 (nϭ16) fmol/minϫcm 2 at 0 shear stress to 5.70Ϯ2.51 (nϭ5) and 0.54Ϯ0.54 (nϭ4) fmol/minϫcm 2 at shear stresses of 0.68 and 13.2 dyne/cm 2 , respectively. At the same time, immunogold labeling showed that TF antigen on the endothelial surface increased Ͼ5-fold with shear stress, whereas TFPI antigen on the surface increased 2-fold. The secretion of TFPI (appearance of new supernatant TFPI) rose from 7.4Ϯ2.4 (nϭ12) ϫ10 Ϫ3 fmol/minϫcm 2 at 0 shear stress to 23.7Ϯ7.3 (nϭ9) and 50.2Ϯ14.3 (nϭ4) ϫ10 Ϫ3 fmol/minϫcm 2 at 0.68 and 13.2 dyne/cm 2 , respectively. TF mRNA amplification products were not markedly changed by shear stress. We conclude that acute application of shear stress reduces functional, but not antigenic, expression of TF by intact, activated endothelial cell monolayers in a manner associated with shear stress-augmented endothelial cell secretion of TFPI. Key Words: tissue factor Ⅲ endothelial cells Ⅲ shear stress Ⅲ tissue factor pathway inhibitor T issue factor (TF) is the membrane-bound glycoprotein that serves as the nonenzymatic cofactor for factor VII/VIIa in the initiation of blood coagulation, whereas TF pathway inhibitor (TFPI), its principal inhibitor, binds to both activated factor X and the ternary complex formed by TF, factor VIIa, and factor Xa. Both TF and TFPI are synthesized by endothelial cells (ECs), the former usually only after cell activation. 1 With regard to increasing shear stress, functional TF has been shown to increase over a wide range of shear stress for a noncellular system of lipid bilayers, to which has been added exogenous TF that has been immobilized on the inner surface of a glass tube. 2 A similar increase has been demonstrated for monolayers of human fibroblasts, 3 which express TF in the absence of cytokine activation. With respect to monolayers of activated human ECs, however, the dependence on shear stress is more complex, passing through a maximum with respect to shear stress under 1 set of conditions 3 or decreasing monotonically with shear stress under another. 4 Prior work h...

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Section: Discussionsupporting

confidence: 77%

Section: Discussionmentioning

confidence: 53%

Section: Anti-tfpi Assaymentioning

confidence: 99%

Section: Anti-tfpi Assaymentioning

confidence: 99%

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Shear Stress Decreases Endothelial Cell Tissue Factor Activity by Augmenting Secretion of Tissue Factor Pathway Inhibitor

Grabowski

Reininger

Petteruti

et al. 2001

ATVB

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“…A new form of bronchopulmonary dysplasia (BPD) was first described in 1999 [1,2] and is characterized by an arrest of lung development [2]. The disruption of lung growth observed in “new BPD” is associated with pulmonary vascular disease in which decreased pulmonary blood vessel growth combined with functional and structural vascular abnormalities have been suggested to decrease the area for gas exchange, finally resulting in pulmonary hypertension (PH) [3].…”

Section: Introductionmentioning

confidence: 99%

An Echocardiographic Screening Program Helps to Identify Pulmonary Hypertension in Extremely Low Birthweight Infants with and without Bronchopulmonary Dysplasia: A Single-Center Experience

Mehler

Cate²,

Keller³

et al. 2017

Neonatology

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Background: Pulmonary hypertension (PH) affects 1 in 6 infants with a birthweight <1,000 g (extremely low birthweight; ELBW) and is frequently associated with bronchopulmonary dysplasia (BPD). If untreated, the mortality rates of the disease are high. Objectives: The aim of this study was to characterize risk factors for PH in ELBW infants and to describe the timing of onset of the disease by setting up a screening program. Methods: ELBW infants treated at the Department of Neonatology (level III neonatal intensive care unit at the University of Cologne Medical Centre, Germany) between January 2010 and March 2015 were included. Echocardiography screening for PH was performed either before discharge or if BPD was diagnosed. Additionally, infants had at least 1 echocardiographic scan after discharge. Survival with PH, age at diagnosis of PH, and risk factors associated with PH were assessed. Results: In total, 34/188 (18%) infants had PH. Of these, 14 (41%) were identified after discharge. Another 11 (32%) were diagnosed with PH without suffering from moderate or severe BPD. The risk factors for diagnosis of PH were moderate (odds ratio, OR 4 [2-8]) or severe BPD (OR 13 [2-71]), prolonged rupture of membranes >7 days (OR 5 [1-19]), and birthweight below the 3rd percentile (OR 3 [1-9]). All infants with PH before discharge and 50% diagnosed after discharge were treated with sildenafil (2.0 mg/kg/day). PH resolved and sildenafil was discontinued in all patients after a median duration of 13 months (IQR 8-20). Conclusions: An echocardiographic screening program may help to identify infants with PH. Examinations should include all ELBW infants irrespective of the presence of BPD and be continued after discharge.

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Evaluation of tissue-factor production by human endothelial cells incubated with three acrylic bone cements

Cenni

Ciapetti

Granchi

et al. 2001

J. Biomed. Mater. Res.

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The effect of three methacrylate-based cements used for the fixation of joint prostheses on tissue-factor production by human umbilical vein endothelial cells was evaluated in vitro. The extracts in the culture medium of the cements were tested after 1-h and 7-day curing. The endothelial cells were incubated with the cement extracts for 4 h, and then the tissue factor was determined in cell lysates with both the recalcification time and enzyme immuno assay. The cements did not induce significant production of tissue factor and, therefore, did not activate the extrinsic pathway of coagulation within the limits of the mechanism considered.

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Comparison of Tissue Factor Pathway in Human Umbilical Vein and Adult Saphenous Vein Endothelial Cells: Implications for Newborn Hemostasis and for Laboratory Models of Endothelial Cell Function

Cited by 149 publications

References 23 publications

Shear Stress Decreases Endothelial Cell Tissue Factor Activity by Augmenting Secretion of Tissue Factor Pathway Inhibitor

Shear Stress Decreases Endothelial Cell Tissue Factor Activity by Augmenting Secretion of Tissue Factor Pathway Inhibitor

An Echocardiographic Screening Program Helps to Identify Pulmonary Hypertension in Extremely Low Birthweight Infants with and without Bronchopulmonary Dysplasia: A Single-Center Experience

Evaluation of tissue-factor production by human endothelial cells incubated with three acrylic bone cements

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