Key Laboratory of Biorheological Science and Technology (Chongqing University), Ministry of Education, Chongqing Engineering Laboratory in Vascular Implants, Bioengineering College of Chongqing University, Chongqing 400044, China.
J Biomed Mater Res A. 2011 Jun 15;97(4):423-32. doi: 10.1002/jbm.a.33066. Epub 2011 Apr 11.
The aim of this study is to construct a biocompatible coating of a drug-eluting stent through the incorporation of chitosan with monoclonal antibody (mAb) to a platelet glycoprotein (GP) IIIa receptor, by electrostatic layer-by-layer (LBL) adsorption of oppositely charged polyelectrolytes and proteins. The platelet maximum aggregation rate and aggregation inhibition rate tests confirm the bioactivity of mAb in different pH assembly environments. The fluorescence spectra test and confocal laser scanning microscopy observation were used to monitor the LBL assembly process of the mAb/chitosan multilayer on the surface of the aminolyzed Poly-L-lactic acid (PLLA) membrane, when using Rhodamine B isothiocyanate-labeled mAb and Fluorescein isothiocyanate-labeled chitosan. The in vitro platelet adhesion experiment demonstrated the amicable blood compatibility of the mAb/chitosan multilayer. The endothelial cell adhesion and migration test revealed that the multilayer could improve the cytocompatibility of the PLLA matrix in terms of cell attachment, proliferation, and migration. An in vitro perfusion circuit was designed to evaluate the release rates measured by a radioisotope technique with ¹²⁵I-labeled GP IIIa mAb. The different eluting curves of the mAb/chitosan-assembled stent and mAb physically absorbed stent showed the improvement of mAb's release character when using LBL self-assembly technology. Our method to prepare a biocompatible stent surface with mAb/chitosan multilayers has proved to be favorable and effective in vitro, thus justifying further evaluation to improve the biocompatibility in an animal model test.
本研究旨在通过静电层层（LBL）吸附带相反电荷的聚合物电解质和蛋白质，将壳聚糖与抗血小板糖蛋白（GP）IIIa 受体的单克隆抗体（mAb）结合，构建一种具有生物相容性的药物洗脱支架涂层。血小板最大聚集率和聚集抑制率测试证实了 mAb 在不同 pH 组装环境下的生物活性。荧光光谱测试和共聚焦激光扫描显微镜观察用于监测 Rhodamine B 异硫氰酸酯标记的 mAb 和荧光素异硫氰酸酯标记的壳聚糖在氨解聚乳酸（PLLA）膜表面的 mAb/壳聚糖多层 LBL 组装过程。体外血小板黏附实验表明 mAb/壳聚糖多层具有良好的血液相容性。内皮细胞黏附和迁移实验表明，多层可提高 PLLA 基质的细胞黏附、增殖和迁移的细胞相容性。设计了一个体外灌注回路，通过放射性同位素技术，用 ¹²⁵I 标记的 GP IIIa mAb 测量释放率。mAb/壳聚糖组装支架和 mAb 物理吸附支架的不同洗脱曲线表明，使用 LBL 自组装技术可改善 mAb 的释放特性。我们用 mAb/壳聚糖多层制备具有生物相容性的支架表面的方法在体外已被证明是有利和有效的，因此有理由进一步评估在动物模型试验中提高生物相容性。
