scholarly journals Effect of luminal surface structure of decellularized aorta on thrombus formation and cell behavior

PLoS ONE ◽  
2021 ◽  
Vol 16 (5) ◽  
pp. e0246221
Author(s):  
Mako Kobayashi ◽  
Masako Ohara ◽  
Yoshihide Hashimoto ◽  
Naoko Nakamura ◽  
Toshiya Fujisato ◽  
...  
Keyword(s):  
Surface Structure ◽  
Blood Clotting ◽  
Heart Valves ◽  
Thrombus Formation ◽  
Functional Expression ◽  
Sem Analysis ◽  
Cell Behavior ◽  
Luminal Surface ◽  
Immunohistological Staining

Due to an increasing number of cardiovascular diseases, artificial heart valves and blood vessels have been developed. Although cardiovascular applications using decellularized tissue have been studied, the mechanisms of their functionality remain unknown. To determine the important factors for preparing decellularized cardiovascular prostheses that show good in vivo performance, the effects of the luminal surface structure of the decellularized aorta on thrombus formation and cell behavior were investigated. Various luminal surface structures of a decellularized aorta were prepared by heating, drying, and peeling. The luminal surface structure and collagen denaturation were evaluated by immunohistological staining, collagen hybridizing peptide (CHP) staining, and scanning electron microscopy (SEM) analysis. To evaluate the effects of luminal surface structure of decellularized aorta on thrombus formation and cell behavior, blood clotting tests and recellularization of endothelial cells and smooth muscle cells were performed. The results of the blood clotting test showed that the closer the luminal surface structure is to the native aorta, the higher the anti-coagulant property. The results of the cell seeding test suggest that vascular cells recognize the luminal surface structure and regulate adhesion, proliferation, and functional expression accordingly. These results provide important factors for preparing decellularized cardiovascular prostheses and will lead to future developments in decellularized cardiovascular applications.

scholarly journals Effect of luminal surface structure of decellularized aorta on thrombus formation and cell behavior

2021 ◽  
Author(s):  
Mako Kobayashi ◽  
Masako Ohara ◽  
Yoshihide Hashimoto ◽  
Naoko Nakamura ◽  
Toshiya Fujisato ◽  
...  
Keyword(s):  
Surface Structure ◽  
Blood Clotting ◽  
Heart Valves ◽  
Thrombus Formation ◽  
Functional Expression ◽  
Sem Analysis ◽  
Cell Behavior ◽  
Luminal Surface ◽  
Immunohistological Staining

AbstractAs the number of cardiovascular diseases increases, artificial heart valves and blood vessels have been developed. Although cardiovascular application using decellularized tissue have been studied, the mechanism of their functionality is still unknown. To find the important factor for preparing decellularized cardiovascular prothesis which shows good in vivo performance, the effect of luminal surface structure of decellularized aorta on thrombus formation and cell behavior was investigated. Various luminal surface structures of decellularized aorta were prepared by heating, drying and peeling. The luminal surface structure and collagen denaturation was evaluated by immunohistological staining, collagen hybridizing peptide (CHP) staining and scanning electron microscopy (SEM) analysis. To evaluate the effect of luminal surface structure of decellularized aorta on thrombus formation and cell behavior, blood clotting test and recellularization of endothelial cells and smooth muscular cells were performed. The results of blood clotting test showed that the closer the luminal surface structure is to native aorta, the higher the anti-coagulant property. From the result of cell seeding-test, it was suggested that vascular cells recognize the luminal surface structure and regulate adhesion, proliferation and functional expression. These results will provide important factor for preparing decellularized cardiovascular prothesis and lead to future development on decellularized cardiovascular applications.

Magnesium Inhibits Arterial Thrombi after Vascular Injury in Rat: In Vivo Impairment of Coagulation

2001 ◽  
Vol 86 (11) ◽  
pp. 1292-1295 ◽  
Author(s):  
Luigi Sironi ◽  
Luca Tedeschi ◽  
Anna Maria Calvio ◽  
Susanna Colli ◽  
Elena Tremoli ◽  
...  
Keyword(s):  
High Frequency ◽  
Blood Clotting ◽  
Magnesium Deficiency ◽  
Ex Vivo ◽  
Thrombus Formation ◽  
Clotting Time ◽  
Chemically Induced ◽  
In Vivo Effects ◽  
Blood Clotting Time

SummaryMagnesium deficiency is associated with a high frequency of cardiac arrhythmia, hypertension and sudden ischemic death. We investigated the in vivo effects of intravenous magnesium administration in a rat model of chemically induced (FeCl3) carotid thrombosis. The infusion of magnesium sulfate (MgSO4) before the topical application of FeCl3 prevented thrombus formation at concentrations of 0.3 M and 0.6 M, and delayed it even at 0.15 M. Similar results were obtained with MgCl2. The infusion of MgSO4 0.6 M seven minutes after FeCl3 application delayed but did not prevent thrombus formation. MgSO4 slightly reduced platelet aggregation ex vivo without affecting plasma clotting tests, but in vivo blood clotting time was markedly prolonged (tail transection method), thus indicating profoundly impaired coagulation. These data provide a rationale for the use of magnesium as an anti-thrombotic agent, but its pharmacological effect critically depends on the timing of administration.

scholarly journals Novel Extracellular x-Prolyl Dipeptidyl-Peptidase (DPP) from Streptococcus gordonii FSS2: an Emerging Subfamily of Viridans Streptococcal x-Prolyl DPPs

2001 ◽  
Vol 69 (9) ◽  
pp. 5494-5501 ◽  
Author(s):  
J. M. Goldstein ◽  
A. Banbula ◽  
T. Kordula ◽  
J. A. Mayo ◽  
J. Travis
Keyword(s):  
Heart Valves ◽  
Thrombus Formation ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Critical Factor ◽  
Dipeptidyl Peptidase ◽  
Streptococcus Gordonii ◽  
Pcr Cloning

ABSTRACT Streptococcus gordonii is generally considered a benign inhabitant of the oral microflora, and yet it is a primary etiological agent in the development of subacute bacterial endocarditis (SBE), an inflammatory state that propagates thrombus formation and tissue damage on the surface of heart valves. Strain FSS2 produced several extracellular aminopeptidase and fibrinogen-degrading activities during growth in culture. In this report we describe the purification, characterization, and cloning of a serine class dipeptidyl-aminopeptidase, an x-prolyl dipeptidyl-peptidase (Sg-xPDPP, for S. gordonii x-prolyl dipeptidyl-peptidase), produced in a pH-controlled batch culture. Purification of this enzyme by anion exchange, gel filtration, and hydrophobic interaction chromatography yielded a protein monomer of approximately 85 kDa, as shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (PAGE) under denaturing conditions. However, under native conditions, the protein appeared to be a homodimer on the basis of gel filtration and PAGE. Kinetic studies indicated that purified enzyme had a unique and stringent x-prolyl specificity that is comparable to both the dipeptidyl-peptidase IV/CD26 and lactococcal x-prolyl dipeptidyl-peptidase families. Nested PCR cloning from an S. gordonii library enabled the isolation and sequence analysis of the full-length gene. A 759-amino-acid polypeptide with a theoretical molecular mass of 87,115 Da and a calculated pI of 5.6 was encoded by this open reading frame. Significant homology was found with the PepX gene family fromLactobacillus and Lactococcus spp. and putative x-prolyl dipeptidyl-peptidases from other streptococcal species. Sg-xPDPP may serve as a critical factor for the sustained bacterial growth in vivo and furthermore may aid in the proteolysis of host tissue that is commonly observed during SBE pathology.

Abstract 13183: A Novel Model of in vivo Platelet Thrombus Imaging Using CD41-ZsGreen1 Transgenic Rats

Circulation ◽  
2015 ◽  
Vol 132 (suppl_3) ◽  
Author(s):  
Makoto Mizuno ◽  
Atsuyuki Tomizawa ◽  
Kousaku Ohno ◽  
Joseph A. Jakubowski ◽  
Atsuhiro Sugidachi
Keyword(s):  
Platelet Aggregation ◽  
Blood Clotting ◽  
Thrombus Formation ◽  
Green Fluorescence Protein ◽  
In Vivo Models ◽  
Thrombus Imaging ◽  
Vehicle Treatment ◽  
Platelet Dynamics ◽  
Thrombotic Diseases

Introduction: Platelets play important roles in both hemostasis and thrombotic diseases including myocardial and cerebral infarction, and stent thrombosis. While mice models of in vivo fluorescence imaging have been described, there are few suitable in vivo models for platelet imaging in rats which are commonly used as thrombosis models. The objective of this study was to generate fluorescent platelets and assess their dynamics in injured arteries in rats. Methods: We generated CD41-ZsGreen1 transgenic (TG) rats, in which the green fluorescence protein ZsGreen1 was expressed specifically in megakaryocytes and thus platelets, and evaluated platelet dynamics in in vivo thrombus imaging experiments. Hematological, including platelet aggregation and blood clotting time, and biochemical parameters were analysed. In the imaging experiments, saphenous arteries of TG rats were injured with 10% FeCl 3 , and thrombus formation was evaluated as fluorescence accumulation using confocal microscopy. The effect of prasugrel, a thienopyridine ADP receptor antagonist, on thrombus formation was also evaluated. Results: The CD41-ZsGreen1 TG rats exhibited normal hematological and biochemical values with the exception of body weight and erythroid parameters, which were significantly lower than those in wild type (WT) rats. Platelet aggregation, induced by 20 μM ADP or 10 μg/mL collagen, and blood clotting times (APTT and PT) were not significantly different between TG and WT rats. In thrombus formation experiments, FeCl 3 application caused a time-dependent increase in fluorescence intensity in the injured artery in vehicle-treated rats. Prasugrel at an oral dose of 3 mg/kg, treated 2 h before the FeCl 3 application, significantly inhibited fluorescence accumulation compared to the vehicle-treatment group (14.9±2.4 vs 5.4±0.4 arbitrary fluorescence unit at 30 min for vehicle and prasugrel groups, respectively, n=8, p=0.0037). Conclusions: CD41-ZsGreen1 TG rats represent a useful animal for the in vivo imaging of platelet participation in thrombus formation and the evaluation of antithrombotic agents.

A Thin Film Nitinol Heart Valve

2005 ◽  
Vol 127 (6) ◽  
pp. 915-918 ◽  
Author(s):  
Lenka L. Stepan ◽  
Daniel S. Levi ◽  
Gregory P. Carman
Keyword(s):  
Thin Film ◽  
Heart Valve ◽  
Heart Valves ◽  
Thrombus Formation ◽  
Prosthetic Heart Valve ◽  
Flow Loop ◽  
Niti Wire ◽  
Tissue Samples

In order to create a less thrombogenic heart valve with improved longevity, a prosthetic heart valve was developed using thin film nitinol (NiTi). A “butterfly” valve was constructed using a single, elliptical piece of thin film NiTi and a scaffold made from Teflon tubing and NiTi wire. Flow tests and pressure readings across the valve were performed in vitro in a pulsatile flow loop. Bio-corrosion experiments were conducted on untreated and passivated thin film nitinol. To determine the material’s in vivo biocompatibility, thin film nitinol was implanted in pigs using stents covered with thin film NiTi. Flow rates and pressure tracings across the valve were comparable to those through a commercially available 19 mm Perimount Edwards tissue valve. No signs of corrosion were present on thin film nitinol samples after immersion in Hank’s solution for one month. Finally, organ and tissue samples explanted from four pigs at 2, 3, 4, and 6 weeks after thin film NiTi implantation appeared without disease, and the thin film nitinol itself was without thrombus formation. Although long term testing is still necessary, thin film NiTi may be very well suited for use in artificial heart valves.

A Thin Film Nitinol Heart Valve

Aerospace ◽  
2004 ◽  
Author(s):  
Lenka Stepan ◽  
Daniel Levi ◽  
Gregory Carman
Keyword(s):  
Thin Film ◽  
Heart Valve ◽  
Heart Valves ◽  
Thrombus Formation ◽  
Prosthetic Heart Valve ◽  
Flow Loop ◽  
Niti Wire ◽  
Tissue Samples

In order to create a less thrombogenic heart valve with improved longevity, a prosthetic heart valve was developed using thin film nitinol (NiTi). A “butterfly” thin film NiTi valve was constructed using a single, elliptical piece of thin film NiTi and a scaffold made from Teflon tubing and NiTi wire. Flow tests and pressure readings across the valve were performed in vitro in a pulsatile flow loop. Biocorrosion experiments were conducted on untreated and passivated thin film nitinol. To determine the material’s in vivo biocompatibility, thin film nitinol was implanted in a pig using a stent covered with thin film NiTi. Flow rates and pressure tracings across the valve were comparable to those through a commercially available 19 mm Perimount Edwards tissue valve. No signs of corrosion were present on samples of thin film nitinol after immersion in Hank’s solution for 1 month. Finally, organs and tissue samples explanted from the pig 17 days after thin film NiTi implantation appeared without disease, and the thin film nitinol itself was without thrombus formation or endothelialization. Although long term testing will be needed, thin film NiTi may be very well suited for use in artificial heart valves.

In vivo repopulation of tissue engineered heart valves

2010 ◽  
Vol 58 (S 01) ◽  
Author(s):  
PM Dohmen ◽  
A Lembcke ◽  
S Holinski ◽  
JP Braun ◽  
W Konertz
Keyword(s):  
Heart Valves ◽  
Tissue Engineered Heart Valves

One Year In-Vivo Functionality of Transvenously Implanted Tissue-Engineered Pulmonary Heart Valves in Sheep

2016 ◽  
Vol 64 (S 02) ◽  
Author(s):  
H. Spriestersbach ◽  
L. Bruder ◽  
B. Sanders ◽  
E. Fioretta ◽  
D. O h-Icí ◽  
...  
Keyword(s):  
Heart Valves ◽  
One Year

Antithrombotic Effects and Bleeding Time Prolongation with Synthetic Platelet GPIIb/llla Inhibitors in Animal Models of Platelet-Mediated Thrombosis

1994 ◽  
Vol 71 (01) ◽  
pp. 095-102 ◽  
Author(s):  
Désiré Collen ◽  
Hua Rong Lu ◽  
Jean-Marie Stassen ◽  
Ingrid Vreys ◽  
Tsunehiro Yasuda ◽  
...  
Keyword(s):  
Platelet Aggregation ◽  
Bleeding Time ◽  
Bolus Injection ◽  
Cell Injury ◽  
Ex Vivo ◽  
Thrombus Formation ◽  
Femoral Vein ◽  
Thrombotic Occlusion ◽  
Antithrombotic Effects

SummaryCyclic Arg-Gly-Asp (RGD) containing synthetic peptides such as L-cysteine, N-(mercaptoacetyl)-D-tyrosyl-L-arginylglycyl-L-a-aspartyl-cyclic (1→5)-sulfide, 5-oxide (G4120) and acetyl-L-cysteinyl-L-asparaginyl-L-prolyl-L-arginyl-glycyl-L-α-aspartyl-[0-methyltyrosyl]-L-arginyl-L-cysteinamide, cyclic 1→9-sulfide (TP9201) bind with high affinity to the platelet GPIIb/IIIa receptor.The relationship between antithrombotic effect, ex vivo platelet aggregation and bleeding time prolongation with both agents was studied in hamsters with a standardized femoral vein endothelial cell injury predisposing to platelet-rich mural thrombosis, and in dogs with a carotid arterial eversion graft inserted in the femoral artery. Intravenous administration of G4120 in hamsters inhibited in vivo thrombus formation with a 50% inhibitory bolus dose (ID50) of approximately 20 μg/kg, ex vivo ADP-induccd platelet aggregation with ID50 of 10 μg/kg, and bolus injection of 1 mg/kg prolonged the bleeding time from 38 ± 9 to 1,100 ± 330 s. Administration of TP9201 in hamsters inhibited in vivo thrombus formation with ID50 of 30 μg/kg, ex vivo platelet aggregation with an ID50 of 50 μg/kg and bolus injection of 1 mg/kg did not prolong the template bleeding time. In the dog eversion graft model, infusion of 100 μg/kg of G4120 over 60 min did not fully inhibit platelet-mediated thrombotic occlusion but was associated with inhibition of ADP-induccd ex vivo platelet aggregation and with prolongation of the template bleeding time from 1.3 ± 0.4 to 12 ± 2 min. Infusion of 300 μg/kg of TP9201 over 60 min completely prevented thrombotic occlusion, inhibited ex vivo platelet aggregation, but was not associated with prolongation of the template bleeding time.TP9201, unlike G4120, inhibits in vivo platelet-mediated thrombus formation without associated prolongation of the template bleeding time.

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