thromboplastin and Aortic-Valve-Stenosis

Diabetes predisposes to aortic stenosis (AS). We aimed to investigate if diabetes affects the expression of selected coagulation proteins and inflammatory markers in AS valves. Twenty patients with severe AS and concomitant type 2 diabetes mellitus (DM) and 40 well-matched patients without DM scheduled for valve replacement were recruited. Valvular tissue factor (TF), TF pathway inhibitor (TFPI), prothrombin, C-reactive protein (CRP) expression were evaluated by immunostaining and TF, prothrombin, and CRP transcripts were analyzed by real-time PCR. DM patients had elevated plasma CRP (9.2 [0.74-51.9] mg/l vs. 4.7 [0.59-23.14] mg/l, p = 0.009) and TF (293.06 [192.32-386.12] pg/ml vs. 140 [104.17-177.76] pg/ml, p = 0.003) compared to non-DM patients. In DM group, TF-, TFPI-, and prothrombin expression within valves was not related to demographics, body mass index, and concomitant diseases, whereas increased expression related to DM was found for CRP on both protein (2.87 [0.5-9]% vs. 0.94 [0-4]%, p = 0.01) and transcript levels (1.3 ± 0.61 vs. 0.22 ± 0.43, p = 0.009). CRP-positive areas were positively correlated with mRNA TF (r = 0.84, p = 0.036). Diabetes mellitus is associated with enhanced inflammation within AS valves, measured by CRP expression, which may contribute to faster AS progression.

Topics: Aged; Aortic Valve; Aortic Valve Stenosis; C-Reactive Protein; Diabetes Complications; Diabetes Mellitus; Disease Progression; Female; Humans; Inflammation; Lipoproteins; Male; Middle Aged; Prothrombin; RNA, Messenger; Thromboplastin

A role of coagulation in the pathogenesis of aortic stenosis (AS) is unknown. The aim of this study was to investigate the fibrin (Fn) presence and its determinants in calcified stenotic aortic valve leaflets. Twenty-one patients with dominant AS and 17 well-matched patients with dominant aortic insufficiency (AI) undergoing aortic valve replacement were studied. Immunofluorescence analysis was performed on decalcified leaflets using antibodies against human Fn and tissue factor (TF). Fn-positive (41.4%) and TF-positive (25.3%) areas were increased in AS valves compared with AI valves (7.9% and 5.9%, respectively, both p<0.001). Patients with AS had elevated plasma D-dimer (236.4 ± 28 ng/ml, p=0.002) and prothrombin fragment 1+2 (F1.2) (261.7 ± 27.1 pM, p=0.005) compared to AI subjects (142.8 ± 10 ng/ml and 131.2 ± 1.3 pM, respectively). In AS patients Fn-positive areas correlated with TF-positive areas (r=0.68, p=0.0005), D-dimer (r=0.45, p=0.018), F1.2 (r=0.64, p=0.002), the time required for plasma fibrin clot formation (r=0.44, p=0.015) and maximum absorbance of fibrin clots (r=-0.38, p<0.0001), but not with clot permeability or lysis time. Thickness of Fn layer within AS valves was associated with maximum transvalvular gradient (r =0.41, p=0.048). Patients with maximal gradient above 75 mmHg (n=11) showed significant associations between Fn-positive area and both maximal (r =0.63) and mean (r =0.67) transvalvular gradients. Large fibrin amounts, mostly co-localised with TF, are present within the valve leaflets of patients with advanced AS. In vivo thrombin generation and fibrin clot formation are associated with the extent of Fn presence within leaflets, which might contribute to the AS progression.

Topics: Aged; Aortic Valve; Aortic Valve Insufficiency; Aortic Valve Stenosis; Biomarkers; Blood Coagulation; Calcinosis; Chi-Square Distribution; Female; Fibrin; Fibrin Fibrinogen Degradation Products; Fluorescent Antibody Technique; Humans; Male; Middle Aged; Peptide Fragments; Poland; Prothrombin; Severity of Illness Index; Thrombin; Thromboplastin; Ultrasonography

In our previous studies, we showed that a significant proportion of patients with various cardiovascular diseases have active tissue factor (TF) and factor (F)XIa in their plasma. The objective of the present study was to evaluate these two proteins in plasma from patients with aortic stenosis and establish their relationship with the severity of the disease. Fifty-four consecutive patients with aortic stenosis, including 38 (70.4%) severe aortic stenosis patients, were studied. Plasma FXIa and TF activity were determined in clotting assays by measuring the response to inhibitory monoclonal antibodies. TF activity was detectable in plasma from 14 of 54 patients (25.9%), including 13 of 38 with severe aortic stenosis (34.2%) and one of 16 (6.25%) with moderate aortic stenosis (P=0.052). FXIa activity was found in 12 (22.2%) patients, mostly in individuals with severe aortic stenosis (11 of 38, 28.9%, P=0.067). All 12 patients with circulating FXIa had active TF in their plasma as well. Severe aortic stenosis patients with detectable TF had higher maximal (111±20 vs. 97±16 mmHg, P=0.02) and mean (61±12 vs. 53±8 mmHg, P=0.02) transvalvular gradient, compared with those without such activity in plasma. In severe aortic stenosis patients with detectable active TF, prothrombin fragment 1.2, a thrombin generation marker, was higher than that in patients without TF (375±122 vs. 207±64 pM, P<0.001). Detectable FXIa and TF activity was observed for the first time in aortic stenosis patients, primarily in severe ones. This activity correlates with thrombin generation in those patients.

Topics: Adult; Aged; Aged, 80 and over; Aortic Valve Stenosis; Blood Coagulation; Blood Coagulation Tests; Factor XIa; Female; Humans; Male; Middle Aged; Peptide Fragments; Prothrombin; Severity of Illness Index; Thrombin; Thromboplastin; Ultrasonography

We recently demonstrated in an experimental model the expression of tissue factor (TF) in aortic valves. Thrombin, generated at the end of the TF-initiated coagulation cascade, has been shown to cleave the anti-calcific osteopontin (OSP) liberating the pro-inflammatory OSP N-half.. We hypothesized that TF might play an important role in calcific aortic valve stenosis (AS) through thrombin generation and hence evaluated the valvular expression of TF and its inhibitor (TF pathway inhibitor), α-thrombin, OSP and its thrombin-cleaved form (OSP N-half).. Calcified aortic valves were obtained from patients undergoing valve replacement. Protein expression was evaluated by immunostaining and measured using ELISA kits. Transcripts were analyzed by RT-PCR.. We included 52 patients (31 men; age 70 ± 10 years; aortic valve area index 0.35 ± 0.13 cm(2)/m(2)). Immunohistochemistry revealed that TF, OSP and α-thrombin expressions were associated with calcifications at the aortic side of the leaflets. There was an overexpression in calcified regions as compared to non-calcified zones of TF (733.3 ± 70.5 pg/mg vs. 429.4 ± 73.2 pg/mg; p<0.0001), OSP (88.9 ± 12.7 ng/mg vs. 15.0 ± 3.3 ng/mg; p<0.0001) and OSP N-half (0.41 ± 0.06 pmol/mg vs. 0.056 ± 0.011 pmol/mg; p<0.0001). Additionally, both TF and α-thrombin expressions were associated with OSP N-half (r=0.52; p<0.0001 and r=0.33; p=0.019, respectively).. Aortic leaflet TF and α-thrombin expressions and their association with the thrombin-cleaved form of OSP, are a new and important feature of AS. We hypothesize that TF may be involved in the mineralization process of aortic valves by enhancing the generation of the pro-inflammatory OSP N-half through thrombin induction. This pathway deserves further studies to address its implication in the aortic valve calcification process.

Topics: Aged; Aged, 80 and over; Aortic Valve; Aortic Valve Stenosis; Atherosclerosis; Calcinosis; Female; Gene Expression Profiling; Humans; Male; Middle Aged; Osteopontin; Thrombin; Thromboplastin

Topics: Animals; Aortic Valve Stenosis; Aortitis; Calcinosis; Humans; Osteoprotegerin; RANK Ligand; Thromboplastin; von Willebrand Diseases

The role of blood coagulation in the pathogenesis of aortic stenosis (AS) is unknown. Recently, tissue factor (TF) expression in stenotic aortic valves has been reported in animal model.. The aim of the study was to investigate TF expression in valve leaflets obtained from AS patients and to determine its associations with circulating coagulation markers and echocardiographic variables.. We studied 20 patients (10 men, 10 women) with dominant AS (age 62.9 +/-9.6, years, mean gradient 43.62 +/-14.62 mmHg), and 20 well-matched patients with dominant aortic insufficiency (AI) undergoing elective aortic valve replacement. Immunofluorescence was measured on decalcified leaflets using antibodies against human TF and macrophages. Prothrombin fragment 1+2 (F1+2) and circulating TF were determined in plasma prior to surgery.. AS valves were characterized by an increased (all, p <0.001) percentage of TF-positive (24.6%) and macrophage-containing (27.3%) areas detected mainly on the aortic side of the leaflets, compared with AI valves (6.3% and 7.4%, respectively). Patients with AS had elevated F1+2 (262.1 +/-27.8 pmol/l, p <0.001) and plasma TF (median 131.8, interquartile range [91.42-310.56] pg/ml, p = 0.018) compared with AI subjects (136.1 +/-11.9 pmol/l, 65.38 [49.51-87.81] pg/ml, respectively). Percentage of TF-positive areas correlated with plasma TF (r = 0.68, p <0.0001), but not with F1+2. Maximum transvalvular gradient >75 mmHg, but not the aortic valve area, showed associations with percentage of TF-positive areas (r = 0.88, p = 0.0039).. This study is the first full-length report demonstrating the presence of TF associated with macrophage infiltration in human aortic valve leaflets in AS patients.

Topics: Aged; Aortic Valve Stenosis; Blood Coagulation Factors; Female; Gene Expression; Humans; Male; Middle Aged; Thromboplastin; Ultrasonography

This study aims to reveal the morphological, histological, and immunohistochemical mechanism of pannus formation using resected pannus tissue from patients with prosthetic valve dysfunction.. Eleven patients with prosthetic valve (St Jude Medical valve) dysfunction in the aortic position who underwent reoperation were studied. We used specimens of resected pannus for histological staining (hematoxylin and eosin, Grocott's, azan, elastica van Gieson) and immunohistochemical staining (transforming growth factor-beta, transforming growth factor-beta receptor 1, alpha-smooth muscle actin, desmin, epithelial membrane antigen, CD34, factor VIII, CD68KP1, matrix metalloproteinase-1, matrix metalloproteinase-3, and matrix metalloproteinase-9).. Pannus without thrombus was observed at the periannulus of the left ventricular septal side; it extended into the pivot guard, interfering with the movement of the straight edge of the leaflet. The histological staining demonstrated that the specimens were mainly constituted with collagen and elastic fibrous tissue accompanied by endothelial cells, chronic inflammatory cells infiltration, and myofibroblasts. The immunohistochemical findings showed significant expression of transforming growth factor-beta, transforming growth factor-beta receptor 1, CD34, and factor VIII in the endothelial cells of the lumen layer; strong transforming growth factor-beta receptor 1, alpha-smooth muscle actin, desmin, and epithelial membrane antigen in the myofibroblasts of the media layer; and transforming growth factor-beta, transforming growth factor-beta receptor 1, and CD68KP1 in macrophages of the stump lesion.. Pannus appeared to originate in the neointima in the periannulus of the left ventricular septum. The structure of the pannus consisted of myofibroblasts and an extracellular matrix such as collagen fiber. The pannus formation after prosthetic valve replacement may be associated with a process of periannular tissue healing via the expression of transforming growth factor-beta.

Topics: Actins; Activin Receptors, Type I; Aged; Antigens, CD; Aortic Valve; Aortic Valve Stenosis; Cell Division; Echocardiography, Doppler; Endothelium, Vascular; Female; Fibroblasts; Giant Cells, Foreign-Body; Heart Atria; Heart Septum; Heart Valve Prosthesis; Heart Ventricles; Humans; Immunohistochemistry; Japan; Macrophages; Male; Matrix Metalloproteinases; Middle Aged; Mucin-1; Prosthesis Design; Prosthesis Failure; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type I; Receptors, Transforming Growth Factor beta; Reoperation; Thromboplastin; Thrombosis; Transforming Growth Factor beta

Tissue-factor pathway inhibitor (TFPI) inhibits the procoagulant activity of the tissue-factor/factor VIIa complex. It was recently reported that TFPI prevented restenosis following tissue injury in a rabbit atherosclerotic model. In order to clarify the mechanism behind this successful prevention of restenosis, we investigated the direct effect of human recombinant TFPI (h-rTFPI) on the proliferation of cultured human neonatal aortic smooth muscle cells (hSMC). We found that h-rTFPI exhibits inhibitory activity toward hSMC proliferation, while h-rTFPI-C which lacks the carboxyl (C)-terminal region does not. Furthermore, we found that h-rTFPI binds to hSMCs with K(d) = 526 nM but that this binding is inhibited by the addition of the synthetic C-terminal peptide, Lys254-Met276, to h-rTFPI. Thus, the interaction of h-rTFPI with hSMCs mediated via the C-terminal region is responsible for the anti-proliferative action of h-rTFPI. On the basis of these results, we presume that the anti-proliferative effect of h-rTFPI in addition to its anticoagulant function plays a significant role in preventing restenosis following tissue injury.

Topics: Amino Acid Sequence; Aorta; Aortic Valve Stenosis; Binding, Competitive; Cell Division; Cells, Cultured; Dose-Response Relationship, Drug; Heparin; Humans; Infant, Newborn; Lipoproteins; Molecular Sequence Data; Muscle, Smooth, Vascular; Peptide Fragments; Protein Binding; Recombinant Proteins; Serine Proteinase Inhibitors; Thromboplastin