Zhao Jing, Bai Lingchuang, Muhammad Khan, Ren Xiang-Kui, Guo Jintang, Xia Shihai, Zhang Wencheng, Feng Yakai
School of Chemical Engineering and Technology, Tianjin University, Yaguan Road 135, Tianjin 300350, China.
Collaborative Innovation Center of Chemical Science and Chemical Engineering (Tianjin), Weijin Road 92, Tianjin 300072, China.
ACS Biomater Sci Eng. 2019 Jun 10;5(6):2846-2857. doi: 10.1021/acsbiomaterials.9b00431. Epub 2019 May 22.
Blood-contacting materials with antiplatelet aggregation and anticoagulant properties are in great demand in biomedical field. Herein, hydrophilic poly(ethylene glycol) (PEG) and antithrombotic peptide ACH were coimmobilized onto Au to develop a multifunctional surface with remarkable hemocompatibility, antiprotein adsorption, antiplatelet aggregation, anticoagulant properties, and good histocompatibility. PEG can not only help the surface resist nonspecific protein adsorption and improve its hemocompatibility but also be a benefit for the immobilized ACH to maintain its bioactivity in plasma. The anticoagulant peptide ACH can endow the surface with the ability of activated coagulation factor Xa (FXa) inhibition and antiplatelet aggregation activities. Au was first aminated with 2-aminoethanethiol through strong Au-S bond, and then it was reacted with PEG-ACH through active ester chemistry to prepare the multifunctional surface Au-PEG-ACH, among which the heterobifunctional PEG served as a linker to immobilize antithrombotic ACH. The surface chemical structure and composition quantified by attenuated total reflection Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy convinced us of the successful preparation of the PEG-ACH modified Au surface. The surface micromorphology and topography of the modified surfaces were observed by field emission scanning electron microscopy and atomic force microscopy. The good hydrophilicity of the modified Au surface was confirmed by water contact analysis. Enzyme immunoassay analysis demonstrated that the activation level of FXa in plasma after incubation with Au-PEG-ACH was obviously lower than that in control groups. In vitro hemocompatibility evaluation, including hemolysis rate, denaturation of adsorbed fibrinogen, and platelet adhesion and activation, indicated that Au-PEG-ACH possessed good hemocompatibility. In vivo subcutaneous implantation assay also confirmed the milder tissue response of Au-PEG-ACH. These results indicated that the multifunctional surface has great potential for biomedical application.
具有抗血小板聚集和抗凝特性的血液接触材料在生物医学领域有巨大需求。在此，将亲水性聚乙二醇（PEG）和抗血栓肽ACH共固定在金上，以开发一种具有卓越血液相容性、抗蛋白吸附、抗血小板聚集、抗凝特性和良好组织相容性的多功能表面。PEG不仅有助于表面抵抗非特异性蛋白吸附并改善其血液相容性，还有利于固定化的ACH在血浆中保持其生物活性。抗凝肽ACH可赋予表面抑制活化凝血因子Xa（FXa）的能力和抗血小板聚集活性。金首先通过强Au-S键与2-氨基乙硫醇胺化，然后通过活性酯化学与PEG-ACH反应制备多功能表面Au-PEG-ACH，其中异双功能PEG作为连接体固定抗血栓ACH。通过衰减全反射傅里叶变换红外光谱和X射线光电子能谱对表面化学结构和组成进行定量分析，使我们确信成功制备了PEG-ACH修饰的金表面。通过场发射扫描电子显微镜和原子力显微镜观察修饰表面的微观形貌和表面形貌。通过水接触分析证实了修饰的金表面具有良好的亲水性。酶免疫分析表明，与Au-PEG-ACH孵育后血浆中FXa的活化水平明显低于对照组。包括溶血率、吸附纤维蛋白原变性以及血小板粘附和活化在内的体外血液相容性评估表明，Au-PEG-ACH具有良好的血液相容性。体内皮下植入试验也证实了Au-PEG-ACH的组织反应较轻。这些结果表明，该多功能表面在生物医学应用方面具有巨大潜力。
