Istanbul Technical University, Molecular Biology-Genetics and Biotechnology Program (MOBGAM), 34469 Maslak, Istanbul, Turkey.
J Biomater Sci Polym Ed. 2011;22(11):1443-57. doi: 10.1163/092050610X510551. Epub 2010 Jun 30.
The aim of this study is to increase the blood compatibility of polytetrafluoroethylene (PTFE), one of the preferred materials for soft-tissue application, by a two-step procedure: first, the surface was activated by hydrogen plasma followed by acrylamide attachment and, secondly, hirudin, a potent antithrombogenic protein from leeches, was immobilized to the surface. Plasma treatment conditions were optimized and different surfaces were characterized by water contact angle measurements, ATR-FT-IR and X-ray photoelectron spectroscopy (XPS). It was seen that the contact angle of the PTFE decreased from 126° to 55° in optimum conditions. Acrylamide (25% (w/v) in ethanol/acetone (50%, v/v)) was grafted to the surface by the help of argon plasma treatment (1 min, 50 W, 13 Pa). The water contact angle was further decreased to 33° with acrylamide grafting and amide groups, which were subsequently used in protein immobilization, and could be detected both by ATR-FT-IR and XPS analysis. In the second part, hirudin was attached to these amide groups on PTFE surface by an optimized EDC/NHS activation procedure. Then a thrombogenicity test was done to detect hirudin activity. The results showed that there is a significant decrease in the clot formation compared with the untreated PTFE samples and ca. 0.3-0.4 ATU/cm(2) (22-29 ng/cm(2)) of hirudin was enough to prevent the clot formation. A preliminary study showed that the hirudin immobilized membranes keep their antithrombogenic activity for at least 40 days in 37°C in PBS (0.1 M, pH 7.4). As a result, the blood compatibility of PTFE surfaces was ameliorated by plasma-induced monomer grafting and hirudin immobilization, and an alternative material was obtained to be used in medical applications such as vascular grafts, catheters, etc.
本研究旨在通过两步法提高聚四氟乙烯(PTFE)的血液相容性,PTFE 是软组织应用的首选材料之一:首先,通过氢等离子体对表面进行活化,然后接枝丙烯酰胺,其次,将水蛭来源的强效抗凝血蛋白水蛭素固定在表面。优化了等离子体处理条件,并通过水接触角测量、衰减全反射傅里叶变换红外光谱(ATR-FT-IR)和 X 射线光电子能谱(XPS)对不同表面进行了表征。结果表明,在最佳条件下,PTFE 的接触角从 126°降至 55°。借助氩等离子体处理(1 分钟、50 W、13 Pa),丙烯酰胺(25%(w/v)乙醇/丙酮(50%,v/v))接枝到表面。进一步通过丙烯酰胺接枝降低水接触角至 33°,形成酰胺基团,随后可用于蛋白质固定化,ATR-FT-IR 和 XPS 分析均可检测到酰胺基团。在第二部分,通过优化的 EDC/NHS 活化程序将水蛭素固定到 PTFE 表面的这些酰胺基团上。然后进行血栓形成试验以检测水蛭素的活性。结果表明,与未处理的 PTFE 样品相比,凝血形成显著减少,约 0.3-0.4 ATU/cm(2)(22-29ng/cm(2))的水蛭素足以防止凝血形成。初步研究表明,在 37°C 的 PBS(0.1 M,pH 7.4)中,固定有水蛭素的膜至少可保持 40 天的抗血栓形成活性。因此,通过等离子体诱导单体接枝和水蛭素固定化改善了 PTFE 表面的血液相容性,为血管移植物、导管等医疗应用提供了一种替代材料。