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使用光纤显微镜评估急性血小板在不透明金属和聚合物生物材料上的黏附情况。

Assessing acute platelet adhesion on opaque metallic and polymeric biomaterials with fiber optic microscopy.

作者信息

Schaub R D, Kameneva M V, Borovetz H S, Wagner W R

机构信息

Department of Bioengineering, University of Pittsburgh, 420 Center for Biotechnology and Bioengineering, 300 Technology Drive, Pittsburgh, Pennsylvania 15219, USA.

出版信息

J Biomed Mater Res. 2000 Mar 15;49(4):460-8. doi: 10.1002/(sici)1097-4636(20000315)49:4<460::aid-jbm4>3.0.co;2-3.

Abstract

The degree of platelet adhesion and subsequent thrombus formation is an important measure of biocompatibility for cardiovascular biomaterials. Traditional methods of quantifying platelet adhesion often are limited by the need for direct optical access, limited spatial resolution, or the lack of temporal resolution. We have developed a new imaging system that utilizes fiber optics and fluorescence microscopy for the quantification of platelet adhesion. This fiber optic remote microscope is capable of imaging individual fluorescently labeled platelets in whole blood on opaque surfaces. Using this method, platelet adhesion was quantified on a series of metallic [low-temperature isotropic carbon (LTIC); titanium alloy (Ti); diamond-like carbon (DLC); oxidized titanium alloy (TiO); and polycrystalline diamond (PCD)] and polymeric [woven Dacron (WD)] collagen-impregnated Dacron (HEM), expanded polytetrafluoroethylene (ePTFE), and denucleated ePTFE (dePTFE)] biomaterials designed for use in cardiovascular applications. These materials were perfused with heparinized whole human blood in an in vitro parallel plate flow chamber. Platelet adhesion after 5 min of perfusion ranged from 3.7 +/- 1.0 (dePTFE) to 16.8 +/- 1.5 (WD) platelets/1000 micrometer. The temporal information revealed by these studies provides a comparative measure of the acute thrombogenicity of these materials as well as some insight into their long-term hemocompatibilities. Also studied here were the effects of wall shear rate and axial position on platelet adhesion. A predicted increase in platelet adhesion with increased wall shear rate and a trend toward a decrease in platelet adhesion with increased axial distance was observed with the fiber optic microscope. Future applications for this imaging technique may include the long-term evaluation of thrombosis in blood-contacting devices in vitro and, in animal models, in vivo.

摘要

血小板黏附程度及随后的血栓形成是心血管生物材料生物相容性的一项重要指标。传统的血小板黏附定量方法往往受到直接光学观察需求的限制,空间分辨率有限,或者缺乏时间分辨率。我们开发了一种新的成像系统,该系统利用光纤和荧光显微镜对血小板黏附进行定量。这种光纤远程显微镜能够对不透明表面全血中单个荧光标记的血小板进行成像。使用该方法,对一系列用于心血管应用的金属材料[低温各向同性碳(LTIC);钛合金(Ti);类金刚石碳(DLC);氧化钛合金(TiO);以及多晶金刚石(PCD)]和聚合物材料[编织涤纶(WD)]、胶原浸渍涤纶(HEM)、膨体聚四氟乙烯(ePTFE)和去核膨体聚四氟乙烯(dePTFE)]上的血小板黏附情况进行了定量。在体外平行板流动腔中用肝素化的全人血灌注这些材料。灌注5分钟后的血小板黏附量范围为每1000微米3.7±1.0(dePTFE)至16.8±1.5(WD)个血小板。这些研究揭示的时间信息提供了这些材料急性血栓形成性的比较指标,以及对其长期血液相容性的一些见解。这里还研究了壁面剪切速率和轴向位置对血小板黏附的影响。使用光纤显微镜观察到,随着壁面剪切速率增加,血小板黏附量预计会增加,而随着轴向距离增加,血小板黏附量有减少的趋势。这种成像技术未来的应用可能包括在体外对血液接触装置中血栓形成的长期评估,以及在动物模型中的体内评估。

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