To study the influence of blood flow on platelet interaction with selected biological surfaces, we have developed an ex vivo perfusion chamber system. In the present experiments, deendothelialized pig aorta and collagen Type I bundles from Achilles tendon were exposed to either native or heparinized pig blood for periods of time ranging from 1 to 30 minutes, and for flow rates corresponding to wall shear rates of 106 to 3380 sec-1. On the deendothelialized vessel wall, platelet deposition increased with both exposure time and wall shear rate, reaching a maximum value between 5 and 10 minutes of perfusion. At high shear rates and long exposure time (over 10 minutes), platelet deposition decreased from maximum values, indicating that some platelets were embolized by the flow. Ultrastructure analysis of the specimens showed platelet activation, spreading, and degranulation. Collagen induced a progressive accumulation of platelets following a power type curve of aggregate growth with exposure time without reaching a saturation level, even after long perfusion times (30 minutes) and high wall shear rates (3380 sec-1). In conclusion, the reactivity of the exposed materials and the local shear rate, defined by the blood flow and the patent luminal cross section, regulate platelet deposition to injured vascular wall.
The role of thrombin inhibition in platelet vessel wall interaction and thrombus growth was studied under controlled flow conditions. Natural hirudin and recombinant hirudin (r-hirudin), which are specific thrombin inhibitors, were compared with heparinized blood (1.8 +/- 0.2 U/mL) and Ca(2+)-chelated blood in their potential to inhibit platelet interaction and thrombus growth on two biologic vascular surfaces and one immobilized vessel wall component. The substrates were perfused by flowing blood at shear rates typical of patent and stenosed arteries (212 to 1,690/s) for 5 minutes. Platelet deposition was measured by In- 111-labeled platelets. We found that both natural and r-hirudin have similar effects on platelet-substrate interaction. As compared with heparin, platelet deposition to mildly damaged vessel wall and digested collagen type I was not reduced by hirudin or citrate. However, hirudin and citrate significantly reduced platelet deposition to severely damaged vessel wall (platelets x 10(6)/cm2: 93 +/- 10 in heparinized blood v 50 +/- 7 in blood treated with 100 U/mL r-hirudin). Therefore, thrombus growth on areas of severe wall damage is in part dependent on local thrombin production at the site of vascular damage. We also found that hirudin added to heparinized blood reduced platelet deposition to severely injured wall but not to subendothelium or collagen-coated slides. Hirudin added to citrated blood did not affect platelet deposition. Our study indicates that local thrombin generation at the site of severe injury will induce platelet activation and deposition even in the presence of average therapeutic heparin levels that inhibit blood coagulation.
We have studied the effects of high plasma cholesterol levels on platelet-vessel wall interactions under high shear rate conditions typical of the apex of stenotic arteries (2,600 sec"'). Hypercholesterolemia was induced by feeding rabbits a 0.5% cholesterol-rich diet for 60 days. Platelet deposition was studied by use of an annular perfusion chamber and de-endothelialized abdominal rabbit aortas as substrates. After ingestion of the atherogenic diet, the experimental group of animals developed severe hypercholesterolemia, platelets became more fluid as determined by steady-state fluorescence anisotropy (p<0.05), and red blood cell deformability was decreased (/? The process of thrombosis, especially in the vicinity of advanced atherosclerotic lesions, is complex, involving platelets, coagulation proteins, the condition of the vessel wall, and local blood flow conditions. For example, it is believed that rupture or ulceration of an atherosclerotic plaque may precipitate the growth of platelet and fibrinaceous elements in an already narrowed lumen. 16 The localization of these thrombotic masses may well be modulated by local flow conditions. 7 In this context, we 8 have recently demonstrated that the high shear rate areas, typical of flow at the apex of stenotic regions, are the preferred sites for platelet thrombus formation on the injured vessel wall. Thus, both the degree of vascular damage and local flow conditions are important parameters for the study of clinically relevant thrombosis. Dietary fat intake has also been shown to influence not only the development of atherosclerosis but also platelet function in experimental animal models and humans.<4 -3 Platelets from hyperlipemic patients have an increased cholesterol content in the membrane and show enhanced aggregability, release reaction, and thromboxane formation. 9 -12 However, the relation between plasma lipids and platelets with respect to the process of thrombosis has not been fully explored. While numerous studies have been conducted over the past 10 years demonstrating the mechanisms by which various plasma proteins influence platelet-vessel wall interactions, few studies have considered the role of lipoproteins in this process.In this study, we have used a well-established annular perfusion system for studying the effects of increased plasma levels of cholesterol on platelet deposition and thrombus formation on normal rabbit subendothelium under controlled flow conditions. The results of our study indicate that severe hypercholesterolemia significantly increases platelet-vessel wall interactions, a finding that could be important in the thrombotic events associated with advanced atherosclerosis. Methods Animal ModelAdult New Zealand White rabbits (3.0±0.5 kg body weight; mean±SD) were selected by plasma by guest on May 9, 2018 http://atvb.ahajournals.org/ Downloaded from
We have previously observed that von Willebrand factor (vWF) plays an important role in platelet deposition on subendothelium at low values of wall shear rate (200 to 400 seconds-1). In the present study, we have investigated the mechanism responsible for such a defect in platelet deposition at low shear rates in the absence of vWF. Blood from both normal and von Willebrand's disease (vWD) animals was exposed to de-endothelialized aorta from normal pigs for a range of shear rates (200 to 3,000 seconds-1) and exposure times (three to 30 minutes) in a tubular perfusion chamber. Variations in the method of inhibiting coagulation (none, heparin, citrate, hirudin, and EDTA) and of perfusing blood (in vitro v ex vivo) were compared by determining the influence of wall shear rate and vWF on the deposition of 111In-labeled platelets on subendothelium. Whereas platelet deposition was reduced in the absence of vWF for all experimental variations at high shear rates (greater than 850 seconds-1), a defect was observed at low shear rates only when heparinized blood was exposed by means of an ex vivo perfusion system. Maximum sensitivity of the measurement occurs under ex vivo perfusion conditions due to the reduced ability of platelets to deposit in normal blood when recirculated in vitro. Our results indicate that vWF mediates platelet-vessel wall interaction even at low shear rates and that such effect can only be observed in systems where platelet function is minimally affected by the experimental conditions.
In 36 normolipemic pigs randomized to a 4-week feeding with regular pig chow (n = 18, control group) or chow supplemented with cod liver oil (1 ml/kg per day) (n = 18, treated group), treatment with cod liver oil produced a significant decrease in serum cholesterol, low density lipoprotein cholesterol, and triglycerides. Deep carotid arterial wall injury (media exposed) by balloon angioplasty was associated with less 111In-labeled platelet deposition (24.6 +/- 4.8 x 10(6)/cm2 versus 62.5 +/- 17.0 x 10(6)/cm2, p less than 0.05; difference, -33.8 x 10(6)/cm2; 95% confidence interval [CI], -1.9 x 10(6)/cm2 to -73.9 x 10(6)/cm2) and injury-related vasoconstriction (21.3 +/- 2.2% versus 30.9 +/- 2.9%, p less than 0.05; difference, -9.6%; 95% CI, -2.2% to -17.0%) in the cod liver oil-treated group than in the control group; with mild injury (media not exposed), platelet deposition was low and unchanged (6.2 +/- 0.5 x 10(6)/cm2 versus 7.8 +/- 0.7 x 10(6)/cm2; difference, -1.6 x 10(6)/cm2; 95% CI, -1.1 x 10(6)/cm2 to +4.3 x 10(6)/cm2), but associated vasoconstriction was reduced respectively (16.3 +/- 2.0% versus 23.0 +/- 2.2%, p less than 0.05; difference, -6.7%; 95% CI, -0.6% to -12.8%). When arterial blood from cod liver oil-treated pigs superfused normal aortic media ex vivo, platelet deposition onto the normal aortic media was lower than when arterial blood from control pigs superfused the normal aortic media (43.7 +/- 8.8 x 10(6)/cm2 versus 66.8 +/- 13.0 x 10(6)/cm2, p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.