用SPR生物传感器研究纤维蛋白原在生物医用材料表面？…

材料表面对血浆蛋白的吸附特性，是研究和评价生物医用材料血液相容性的重要依据。本文用自制的表面等离激元（ＳＰＲ）传感器，测量了金膜、磷脂ＤＳＰＣ膜、成都科大Ⅱ型聚氨酯、Ｐｅｌｌｅｔｈａｎｅ２３６３－５５Ｄ）聚氨酯及有机玻璃膜表面对纤维蛋白原的动态吸附特性，在纤维蛋白原溶液浓度为５ｍｇ／ｍｌ的相同条件下，磷脂ＤＳＰＣ膜表面吸附纤维蛋白原的速度最低，饱和吸附浓度也最小（表面浓度为１ｎｇ／ｍｍ＾２）。其次     

用SPR生物传感器研究纤维蛋白原在生物医用材料表面的吸附

材料表面对血浆蛋白的吸附特性 ,是研究和评价生物医用材料血液相容性的重要依据。本文用自制的表面等离激元 (SPR)传感器 ,测量了金膜、磷脂DSPC膜、成都科大Ⅱ型聚氨酯、Pellethane2 36 3 55D聚氨酯及有机玻璃膜表面对纤维蛋白原的动态吸附特性 ,在纤维蛋白原溶液浓度为5mg/ml的相同条件下 ,磷脂DSPC膜表面吸附纤维蛋白原的速度最低 ,饱和吸附浓度也最小 (表面浓度为 1ng/mm2 )。其次是裸金膜 (表面浓度为 3.5ng/mm2 ) ,再其次是成都科大Ⅱ型聚氨酯膜 (表面浓度为 3.8ng/mm2 )和Pellethane 2 36 3 55D聚氨酯 (表面浓度为 4 .3ng/mm2 ) ,吸附速度和吸附量最高的是有机玻璃膜 (表面浓度为 4 .5ng/mm2 )。结果表明 ,材料表面对纤维蛋白原的吸附动力学特性 ,与材料的血液相容性密切相关。表面等离激元技术与本文采用的在金膜上铺展高分子材料的离心铺膜法和LB技术等样品制备技术相结合 ,为生物材料表面对蛋白质吸附特性的实时、动态、原位研究提供了一种新的高灵敏度的方法 ,并可能发展成为一种材料生物相容性的测试和评价的新方法。     

表面等离激元共振(SPR)生物传感器对聚氨酯材料血浆蛋白吸附的实时原位动态研究

应用实验室自行研制的自动扫描式表面激元共振(SPR)生物传感器对三种聚氨酯材料进行了血液蛋白质吸附实验，以传感片上的金膜作为对照材料。同时应用原子力显微镜对金膜和聚氨酯材料的超微结构与材料表面上所吸附的蛋白质进行了表征。实验结果显示，四种材料对纤维蛋白原和IgG的吸附量顺序均为：金膜＞H50—0＞H50—50＞H50—100。T—检验结果表明，金膜对纤维蛋白原和IgG吸附量与三种聚氨酯材料均有显著差别。该结果表明聚氨酯材料的血液相容性明显好于金膜对照材料。     

血浆蛋白质在生物材料表面吸附时的Vroman效应

以吸附时间,血浆稀释倍数,狭窄空间的高度为变量研究纤维蛋白原（Ｆｇ）从血浆吸附到生物材料表面时,会发现Ｆｇ的吸附出现一最大值,此即Ｖｒｏｍａｎ效应,研究证实Ｖｒｏｍａｎ效应并非纤维蛋白原所独有的现象,而是一普遍现象,它反映了血浆中的蛋白质对表面有限位点的竞争吸附。我们综述了影响血浆蛋白南在生物材料表面吸附时的Ｖｒｏｍａｎ疚的实验因素,Ｖｒｏｍａｎ效应的机理以及Ｖｒｏｍａｎ效应对于研究血液－材料相互     

聚环氧乙烷硫代物、聚环氧乙烷或氟甲基接枝聚氨酯材料表面上的纤维蛋白原吸附

在探讨血液与医用材料之间的反应和研制与血液相容的医用材料中,研究蛋白质的吸附作用是非常重要的。众所周知,纤维蛋白不但是一种具有高度表面活性的蛋白质,而且也能产生瞬间粘附作用,即“vroman效应”。作者研究已报道具有较好血液相容性的氟化聚氨酯(Pu-PFDA)、聚环氧乙烷(PEO)接枝的聚氨酯(Pu-PEO)以及进一步硫化的Pu(Pu-PEO-SO_3)等材料表面的纤维蛋白原吸附现象。将Pu材料浸入内含14C-标记纤维蛋白原的牛血浆中,拿起后先用PBS缓冲液冲洗,然后再用2%SDS溶液冲洗。采用放射性     

血浆蛋白分子在单壁碳纳米管无纺膜表面吸附行为的研究

近年来,碳纳米管的独特表面拓扑结构、化学组成和优异的物理性能已经吸引了众多领域的研究兴趣,以生物医学应用为目标的探索性研究正在迅速形成一个新的方向。我们以血液接触环境下的应用为目标,通过扫描电镜观察、表面元素分析、以及利用酶联免疫分析技术,系统研究了与凝血过程密切相关的纤维蛋白原、白蛋白、免疫球蛋白以及新鲜血浆在单壁碳纳米管无纺膜(SWNT膜)表面的吸附行为。实验结果显示,单壁碳纳米管无纺膜对血浆中的纤维蛋白原分子具有强烈的倾向性吸附,对免疫球蛋白也显示出一定的吸附性,但是,对白蛋白分子却几乎不吸附。血浆蛋白分子在SWNT膜表面的吸附行为与其在其它碳材料表面和其它大多数生物材料表面的吸附行为显著不同。SWNT膜对血浆蛋白分子的独特吸附作用有可能对后续的血液细胞响应产生重要影响。     

纤维蛋白原对模拟人工体液中不锈钢腐蚀行为的影响

在模拟人工体液PBS溶液中采用电化学测试技术考察了纤维蛋白原对冠状动脉支架用SUS316L和SUS317L不锈钢腐蚀行为的影响。结果表明：纤维蛋白原的存在增大了介质对材料的浸蚀性。在含纤维蛋白原的溶液中,试样的自腐蚀电位负移,维钝电流密度增大,点蚀击穿电位（Eb)降低。并利用扫描电镜观察了吸附现象,发现吸附的纤维蛋白原呈白色条状,不连续地分布在材料表面。     

等离子体表面改性涤纶材料的血浆蛋白吸附与血小板黏附行为研究

采用等离子体表面接枝改性技术在涤纶 (聚对苯二甲酸乙二醇酯 ,PET)材料表面接枝不同分子量的聚乙二醇 (PEG) ,从表面能与界面自由能的角度分析了血浆蛋白 (纤维蛋白原和白蛋白 )在材料表面的竞争吸附关系 ,结果表明接枝了 PEG长链分子的 PET材料具有优先吸附白蛋白的性质 ,其中接枝 PEG6 0 0 0的 PET优先吸附倾向最明显。预接触白蛋白和纤维蛋白原的 PET材料表面的血小板黏附实验表明 :吸附白蛋白的表面能够显著抑制血小板的黏附和聚集 ,表现出好的血液相容性 ,而吸附了纤维蛋白原的材料表面具有降低血液相容性的性质。     

纳米碳改性聚氨酯复合材料的表面抗凝血性能

研究纳米碳改性聚氨酯聚合材料表面的血液相容性。将经过表面处理的纳米碳分散到聚氨酯体系中，制成聚氨酯／纳米碳复合薄膜。通过血小板荧光标记人全血灌注实验和羊全血体外循环等实验，观察和测定血小板在材料表面的粘附作用以及血液中血红蛋白浓度、纤维蛋白原浓度的变化，探讨纳米碳对聚氨酯抗凝血性能的影响。实验结果显示聚氨酯／纳米碳表面血小板的粘附明显低于单纯聚氨酯对照组：体外循环4h后，血液中血红蛋白浓度、纤维蛋白原浓度的变化程度减小。表明纳米碳与聚氨酯的复合可以提高材料的血液相容性。     

成科大Ⅰ型和ZH—Ⅲ型聚醚型聚氨酯/聚硅氧烷(SPEU/PDMS)共聚物的血液相容性研究

成科大Ⅰ型和ZH—Ⅲ型聚醚型聚氨酯/聚硅氧烷共聚物是采用不同路线合成的、属Cardiothane(原商品名为AVcothane-51)类型的材料。本文用不同的方法综合评价并对比了人和某些动物的血液相容性:用新鲜人血进行动态凝血时间试验;在狗的股动—静脉短路半体内循环试验中,用同位素标记法测定了纤维蛋白原的吸附量,并用扫描电镜观察血小板的吸附和形态变化;用新鲜的兔富血小板进行体外动态血栓形成试验。评价的结果证明成科大Ⅰ型和ZH—Ⅲ型材料的血液相容性与Avcothane—51的相近。     

The kinetics of baboon fibrinogen adsorption to polymers: in vitro and in vivo studies

Fibrinogen adsorption on polymers from blood may mediate or potentiate thrombosis because of its involvement in both the intrinsic clotting system and the formation of platelet aggregates. While the kinetics of fibrinogen adsorption from plasma in vitro have previously been found to be very different on polar and nonpolar surfaces [T. A Horbett, "The kinetics of adsorption of plasma proteins to a series of hydrophilic-hydrophobic copolymers," ACS Org. Coat. Plas. Chem. 40, 642-646 (1979)] the significance of this difference with respect to thrombogenesis in vivo has not been clarified. In this study, the kinetics of deposition of baboon 125I fibrinogen from plasma in vitro or from blood in vivo on a series of polymers was measured. The polymers chosen for this study had previously been found to have a large range in surface polarity and reactivity in the in vivo baboon shunt model. The kinetics of fibrinogen adsorption in vitro were observed to be of three types, depending on the polymer: high initial adsorption decreasing to a lower steady state value; constant throughout the time course; low initial adsorption rising steadily to a plateau value. In vivo, fibrinogen deposition kinetics were of two types: low, constant deposition throughout the time course, independent of heparinization; low deposition initially followed by a second phase of greatly increased deposition (probably as fibrin) which was prevented or greatly decreased by heparinizing the animals. Polymers for which fibrinogen adsorption increased to a plateau in vitro were found to have a heparin inhibitable second phase of enhanced in vivo fibrinogen deposition. These polymers also have been found in previous studies to enhance the rate of platelet destruction when used as in vivo shunts on baboons. Conversely, most polymers with high initial in vitro fibrinogen adsorption followed by a decrease had low fibrinogen deposition behavior in vivo and were also minimally destructive of platelets. The adsorption kinetics of fibrinogen to polymers from blood in vivo and in vitro and the consumption of platelets in vivo induced by the polymers all vary with polymer polarity. More polar polymers had in vitro fibrinogen kinetics characterized by a rise to a plateau, in vivo fibrinogen deposition characterized by a second stage of great increase inhibitable by heparin, and enhanced platelet consumption. The correlation of three separate indicators of surface thrombogenicity with surface polarity suggests that more polar materials may be more thrombogenic because of an influence on the way in which fibrinogen interacts with these surfaces.     

The Vroman effect in tube geometry: the influence of flow on protein adsorption measurements.

The transient adsorption of fibrinogen from plasma (a manifestation of the Vroman effect), due in large part to displacement by trace proteins such as high-molecular-weight kininogen (HK), factor XII, and plasminogen, has traditionally been studied in nonflowing systems in this laboratory. This paper reports new data on adsorption in tubing geometry under laminar flow. Fibrinogen adsorption from human blood plasma and whole blood diluted to varying exents was measured on glass and polyethylene tubing. The presence of flow did not change the nature of the Vroman effect, except that the processes of adsorption and displacement, which are typically diffusion-limited in static systems, were augmented by convective transport. At the highest applied shear rates of 408 and 510 s-1, the initial adsorption rate of fibrinogen was estimated to be 5.0 X 10(-5) cm/s on both surfaces. The intrinsic rate of displacement of fibrinogen (due to the Vroman effect) at high shear rates was about ten times faster from glass than from polyethylene based on data taken 5 min after the experiment started. The rates of fibrinogen adsorption and displacement were not observed to be significantly augmented by the cellular elements of whole blood at dilutions exceeding 20:1. The consistently observed axial dependence of adsorption in static and flow experiments in tubing geometry was investigated. It was concluded that the effect results, under most conditions, from the creation of a concentration boundary layer during the displacement of the equilibrating buffer by the injected protein solution. The possibility of local depletion due to rapid adsorption during injection or the final displacement of the protein solution was concluded to make lesser contributions to axial variations in measured adsorption.     

Plasma-deposited tetraglyme surfaces greatly reduce total blood protein adsorption, contact activation, platelet adhesion, platelet procoagulant activity, and in vitro thrombus deposition

The ability of tetraethylene glycol dimethyl ether (tetraglyme) plasma deposited coatings exhibiting ultralow fibrinogen adsorption to reduce blood activation was studied with six in vitro methods, namely fibrinogen and von Willebrand's factor adsorption, total protein adsorption, clotting time in recalcified plasma, platelet adhesion and procoagulant activity, and whole blood thrombosis in a disturbed flow catheter model. Surface plasmon resonance results showed that tetraglyme surfaces strongly resisted the adsorption of all proteins from human plasma. The clotting time in the presence of tetraglyme surfaces was lengthened compared with controls, indicating a lower activation of the intrinsic coagulation cascade. Platelet adhesion and thrombin generation by adherent platelets were greatly reduced on tetraglyme-coated materials, compared with uncoated and Biospan-coated glass slides. In the in vitro disturbed blood flow model, tetraglyme plasma coated catheters had 50% less thrombus than did the uncoated catheters. Tetraglyme-coated materials thus had greatly reduced blood interactions as measured with all six methods. The improved blood compatibility of plasma-deposited tetraglyme is thus not only due to their reduced platelet adhesion and activation, but also to a generalized reduction in blood interactions.     

Changes in fibrinogen adsorbed to segmented polyurethanes and hydroxyethylmethacrylate-ethylmethacrylate copolymers.

Fibrinogen adsorption from blood to biomaterials may regulate platelet adhesion and thrombus formation because of fibrinogen's central role in the coagulation cascade and its ability to bind specifically to the platelet membrane glycoprotein (GP) IIb-IIIa. Adsorption of fibrinogen from blood plasma to many materials exhibits a maximum with respect to plasma dilution and exposure time (the Vroman effect). In this study fibrinogen adsorption to several polymers was examined to ascertain the influence of controlled changes in surface chemistry on the Vroman effect. The materials included hydroxyethylmethacrylate-ethylmethacrylate (HEMA/EMA) copolymers, Biomer, and a series of segmented polyurethanes (PEUs), two of which contained fluorinated chain extenders. Each material exhibited maximal adsorption of fibrinogen at intermediate plasma concentrations. Little effect of soft-segment type or molecular weight was observed and no significant differences in fibrinogen adsorption to the fluorinated PEUs were seen. Changes in the strength of fibrinogen attachment to these materials with time after adsorption were also assessed. Fibrinogen adsorbed for 1 min was displaced more readily by blood plasma than that adsorbed for 1 h, regardless of the material. The more hydrophobic polymers exhibited greater retention of adsorbed fibrinogen. In addition, the fraction of fibrinogen retained by polyethylene depended on the amount of fibrinogen adsorbed to the surface, being greatest when the surface loading was the least. These studies indicate that spreading or transition of adsorbed fibrinogen molecules from a weakly to tightly bound state is a general consequence of protein adsorption to solid surfaces.     

Quantitative evaluation of interaction force of fibrinogen at well-defined surfaces with various structures

The effects of functional groups and structures at the surface of biomaterials on protein adsorption were examined using direct interaction force measurements. Three kinds of surface structures were evaluated: polymer brushes, self-assembled monolayers with low molecular weight compounds, and surfaces with conventional polymer coatings. These surfaces had various functional groups including phosphorylcholine (PC) group. The surface characterization demonstrated that surface wettability and flexibility depended on both the structure of the surface and the functional groups at the surface. The interactions of protein with these surfaces were evaluated by a force vs. distance curve using an atomic force microscope (AFM). We used fibrinogen as the protein, and the fibrinogen was immobilized on the surface of the AFM cantilever by a conventional technique. It was observed that the interaction force of fibrinogen was strongly related to surface hydrophobic nature and flexibility. That is, the interaction force increased with the increasing hydrophobic nature of the surface. The relationship between the amount of fibrinogen adsorbed on the surface and the interaction force showed good correlation in the range of fibrinogen adsorption from 0 to 250?ng/cm², that is, in a monolayered adsorption region. The interaction force decreased with increasing surface viscoelasticity. The most effective surface for preventing fibrinogen adsorption was the polymer brush surface with phosphorylcholine (PC) groups, that is, poly(2-methacryloyloxyethyl phosphorylcholine) brush. The interaction force of this sample was less than 0.1?nN and the amount of fibrinogen adsorbed on the surface was minimal. It was found that the evaluation of protein adsorption based on the interaction force measurement is useful for low-protein adsorption surfaces. It was demonstrated that an extremely hydrophilic and flexible surface could weaken the protein interactions at the surface, resulting in greater resistance to protein adsorption.     

The effects of temperature and buffer on fibrinogen adsorption from blood plasma to glass

[125I]-Fibrinogen was used to measure the adsorption of fibrinogen from baboon plasma to two types of glass (Pyrex and a borosilicate glass) at 25 and 37 degrees C using two different buffers to dilute the plasma, the first being citrate-phosphate buffered saline (CPBSz) and the second isotonic Tris-saline (TRIS), both pH 7.4. In addition, the effects of hydration conditions, rinsing techniques, and glass-cleaning treatments on fibrinogen adsorption were evaluated. The data reveal that lower temperatures and the use of TRIS to dilute the plasma significantly enhance fibrinogen adsorption to both types of glass. As has been observed in the past, fibrinogen adsorption peaked at intermediate plasma concentrations on both Pyrex and borosilicate glass (the so-called Vroman effect), but almost twice as much fibrinogen adsorbed to glass when TRIS was used to dilute the plasma instead of CPBSz. Moreover, up to five times as much fibrinogen adsorbed to both types of glass at 25 degrees C compared with 37 degrees C. No effects of the rinsing technique or glass-cleaning treatment were observed.     

A model for thromboembolization on biomaterials

A model was developed to describe the kinetics of protein and platelet deposition and embolization on biomaterials. The model assumes that proteins can be adequately represented by fibrinogen, albumin, and Factor XII, that protein adsorption is Langmuir-type, that surfaces are homogeneous, and that all adsorption and deposition steps are first order. Eleven model parameters were determined from literature experimental data from ex vivo experiments utilizing canine and baboon blood on Silastic, one parameter came from adsorption of Factor XII on glass, and three parameters were obtained by minimizing differences between experimental and predicted fibrinogen adsorption, and platelet deposition and embolization behavior. The model well predicted observed behavior for fibrinogen adsorption, platelet deposition, and platelet embolization on Silastic, and platelet embolization from both polyacrylamide and HEMA-MAAC.     

Transient adsorption of fibrinogen on foreign surfaces: similar behavior in plasma and whole blood

The adsorption of fibrinogen from both human whole blood and plasma to a number of "foreign" surfaces is reported. Adsorption was measured as a function of plasma or blood dilution using radioiodine labeling. We showed previously that adsorption of fibrinogen from plasma exhibits a maximum at a plasma dilution of about 100:1, and have attributed this behavior to competition from other plasma proteins. (The same phenomenon is manifest as a time transient in fibrinogen adsorption.) In the present work we show that exactly the same trends are observed in whole blood. For each of the four surfaces, glass, siliconized glass, collagen-coated glass and polyethylene, the adsorption of fibrinogen as a function of dilution is the same in whole blood as in plasma. Each of these surfaces shows a unique dependence of fibrinogen adsorption on plasma or blood dilution. On cuprophane and a hydrophilic polyether urethane there is essentially no adsorption of fibrinogen from blood or plasma. For the hydrophilic polyurethane this result may be artifactual, but the absence of fibrinogen binding to cuprophane in blood or plasma is real since fibrinogen is found to be adsorbed in monolayer amounts from buffer.     

In vitro inhibition of biophysical surface properties and change in ultrastructures of exogenous pulmonary surfactant by albumin or fibrinogen.

In order to observe the effects of serum albumin and fibrinogen on biophysical surface properties and the morphology of pulmonary surfactant in vitro, we measured the surface adsorption rate, dynamic minimum and maximum surface tension (min-, max-ST) by Pulsating Bubble Surfactometer, and demonstrated ultrastructures on a series of mixtures with varying concentrations of albumin or fibrinogen and Surfactant-TA. The albumin and fibrinogen significantly inhibited the adsorption rate and ST-lowering properties of surfactant through increasing STs of adsorption rate, min-ST, and max-ST. The characteristic morphology of the Surfactant-TA changed from lamellar rod-like structure with open ends into spherical structures with loss of their open ends by mixing with albumin or fibrinogen. These inhibitory effects of albumin and fibrinogen on surface properties of surfactant were dependent upon the increasing concentration of albumin or fibrinogen. We concluded that albumin and fibrinogen significantly altered surfactant function and its ultrastructural morphology in vitro. These findings support the concept that albumin and fibrinogen-induced surfactant dysfunction may play an important role in the pathophysiology of adult respiratory distress syndrome, and this adverse effect of albumin and fibrinogen on surfactant might be overcome by administration of large doses of exogenous surfactant.     

Nonfouling biomaterials based on polyethylene oxide-containing amphiphilic triblock copolymers as surface modifying additives: protein adsorption on PEO-copolymer/polyurethane blends

Surface modification of a segmented polyurethane was achieved by blending with novel PEO-containing amphiphilic triblock copolymers (PEO-polyurethane-PEO). Three copolymers having different PEO MW (550, 2000, 5000) were used as surface modification additives. The protein resistance of the blend surfaces was evaluated using radiolabeling methods. On the blends of copolymers with PEO blocks of MW 2000 and 5000, fibrinogen adsorption from physiologic buffer decreased with increasing copolymer content up to 20 wt%. On the blends with PEO blocks of MW 550, resistance to adsorption for a given copolymer content was much greater. For all three blend types at 20% copolymer content, reductions in adsorption compared to the unmodified PU matrix were greater than 95%. Reductions in adsorption were similar for the 20% blends and surfaces prepared by coating the copolymers directly on the matrix, suggesting that the 20% blend surfaces were completely covered by copolymer. At low copolymer content (< or =10 wt %), fibrinogen adsorption decreased with decreasing PEO block length. This was probably due to increasing surface coverage of the copolymers with decreasing block length. It is therefore concluded that surface density of PEO is more important than PEO MW for the protein resistance of these surfaces. Lysozyme, a much smaller protein, showed adsorption trends similar to fibrinogen. The adsorption of fibrinogen and lysozyme from binary solutions to blends of the copolymer with PEO blocks of 2000 MW was investigated to probe the effects of protein size on adsorption resistance. Fibrinogen and lysozyme showed similar fractional decreases in adsorption relative to the PU matrix independent of the surface density of PEO. However lysozyme was enriched in the surface relative to the solution, that is, it was adsorbed preferentially to fibrinogen.