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狒狒纤维蛋白原对聚合材料的吸附及血小板对聚合材料的黏附

Baboon fibrinogen adsorption and platelet adhesion to polymeric materials.

作者信息

Chinn J A, Horbett T A, Ratner B D

机构信息

Department of Chemical Engineering, University of Washington, Seattle 98195.

出版信息

Thromb Haemost. 1991 May 6;65(5):608-17.

PMID:1871724
Abstract

The role of fibrinogen in mediating platelet adhesion to polymers exposed to blood plasma was studied by comparison of the effect of plasma dilution on fibrinogen adsorption and platelet adhesion, and by the use of coagulation factor deficient plasmas. Polyetherurethane substrates were first preadsorbed with dilute plasma, then contacted with washed platelets suspended in a modified, apyrase containing Tyrode's buffer. Platelet adhesion was studied under static conditions in Multiwell dishes, and also under shearing conditions using a parallel plate perfusion chamber. Fibrinogen adsorption and platelet adhesion were measured using 125I radiolabeled baboon fibrinogen and 111In radiolabeled baboon platelets, respectively. Surfaces were characterized by electron spectroscopy for chemical analysis (ESCA). When fibrinogen adsorption to Biomer was measured after 2 h contact with a series of dilute plasma solutions under static conditions, a peak in adsorption was observed from 0.26% plasma, i.e., adsorption was greater from 0.26% plasma than from either more or less dilute plasma. A peak in subsequent platelet adhesion to the plasma preadsorbed surfaces, measured after 2 h static incubation with washed platelets, was also observed but occurred on Biomer preadsorbed with 1.0% plasma. When fibrinogen adsorption was measured after 5 min contact under shearing conditions, the fibrinogen adsorption peak occurred on surfaces that had been exposed to 1.0% plasma. A peak in platelet adhesion to these preadsorbed surfaces, measured after 5 min contact with the platelet suspensions under shearing conditions, was observed on Biomer preadsorbed with 0.1% plasma. Shifts between the positions of the peaks in protein adsorption and platelet adhesion occurred on other polymers tested as well. Platelet adhesion was almost completely inhibited when baboon and human plasmas lacking fibrinogen (i.e., serum, heat defibrinogenated plasma, and congenitally afibrinogenemic plasma) were used. Platelet adhesion was restored to near normal when exogenous fibrinogen was added to fibrinogen deficient plasmas. Adhesion was also inhibited completely when a monoclonal antibody directed against the glycoprotein IIb/IIIa complex was added to the platelet suspension. Platelet adhesion to surfaces preadsorbed to von Willebrand factor deficient plasma was the same as to surfaces preadsorbed with normal plasma. While it appears that surface bound fibrinogen does mediate the initial attachment of platelets to Biomer, the observation that the fibrinogen adsorption and platelet adhesion maxima do not coincide exactly also suggests that the degree of subsequent platelet adhesion is dictated not only by the amount of surface bound fibrinogen but also by its conformation.

摘要

通过比较血浆稀释对纤维蛋白原吸附和血小板黏附的影响,以及使用凝血因子缺乏的血浆,研究了纤维蛋白原在介导血小板黏附于暴露于血浆的聚合物中的作用。首先将聚醚聚氨酯底物用稀释血浆预吸附,然后与悬浮在含有抑肽酶的改良台氏缓冲液中的洗涤血小板接触。在多孔板中于静态条件下研究血小板黏附,也使用平行板灌注室在剪切条件下进行研究。分别使用125I放射性标记的狒狒纤维蛋白原和111In放射性标记的狒狒血小板测量纤维蛋白原吸附和血小板黏附。通过化学分析电子能谱(ESCA)对表面进行表征。当在静态条件下与一系列稀释血浆溶液接触2小时后测量纤维蛋白原对Biomer的吸附时,在0.26%血浆中观察到吸附峰值,即0.26%血浆中的吸附量大于更稀或更浓血浆中的吸附量。在用洗涤血小板进行2小时静态孵育后测量的随后血小板对血浆预吸附表面的黏附峰值也被观察到,但出现在预吸附1.0%血浆的Biomer上。当在剪切条件下接触5分钟后测量纤维蛋白原吸附时,纤维蛋白原吸附峰值出现在暴露于1.0%血浆的表面上。在用血小板悬浮液在剪切条件下接触5分钟后测量的对这些预吸附表面的血小板黏附峰值,在预吸附0.1%血浆的Biomer上被观察到。在测试的其他聚合物上也发生了蛋白质吸附和血小板黏附峰值位置之间的变化。当使用缺乏纤维蛋白原的狒狒和人血浆(即血清、热去纤维蛋白原血浆和先天性无纤维蛋白原血症血浆)时,血小板黏附几乎完全被抑制。当向缺乏纤维蛋白原的血浆中添加外源性纤维蛋白原时,血小板黏附恢复到接近正常水平。当向血小板悬浮液中添加针对糖蛋白IIb/IIIa复合物的单克隆抗体时,黏附也被完全抑制。血小板对预吸附于血管性血友病因子缺乏血浆的表面的黏附与对预吸附正常血浆的表面的黏附相同。虽然表面结合的纤维蛋白原似乎确实介导了血小板与Biomer的初始附着,但纤维蛋白原吸附和血小板黏附最大值不完全一致的观察结果也表明,随后血小板黏附的程度不仅由表面结合的纤维蛋白原的量决定,还由其构象决定。

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