Protein and bacterial fouling characteristics of peptide and antibody decorated surfaces of PEG-poly(acrylic acid) co-polymers
Protein and bacterial fouling characteristics of peptide and antibody decorated surfaces of PEG-poly(acrylic acid) co-polymers
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DOI：
10.1016/j.biomaterials.2003.09.020
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发表时间：
2004-05-01
期刊：
影响因子：
14
通讯作者：
Bryers, JD
中科院分区：
文献类型：
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作者：
Wagner, VE;Koberstein, JT;Bryers, JD
研究了基础聚乙二醇接枝聚丙烯酸 PEG-g-PA 共聚物和表面改性 PEG-g-PA 材料抑制随机蛋白质污染和细菌粘附的潜力。合成的 PEG-g-PA 共聚物可抑制非特异性蛋白质和细胞粘附。然后用细胞粘附肽（YRGDS、YEILDV）或单核细胞/巨噬细胞整合素受体抗体片段（Anti-VLA4、Anti-beta(1)、Anti-beta(2) 和 Anti-CD64）对 PEG-g-PA 共聚物进行共价修饰，这些受体已知可增强巨噬细胞粘附，并可能调节其激活。本工作中生产的材料通过以下方法进行表征：通过接触角测定疏水性；角分辨 X 射线光电子能谱可确认本体材料和表面中是否存在 PEG；水合度；差示扫描量热法；为了评估各种改性表面的防污功效，对三种蛋白质：人血清白蛋白、人纤连蛋白（组分 I）和人免疫球蛋白进行了 I-125 标记。将基础 PEG-g-PA 和用各种肽修饰的 PEG-g-PA 样品在 37℃ 下暴露于含有 2 或 200 μg/ml 一种标记蛋白的溶液中 24 小时。用直接结合的肽修饰的 PEG-g-PA 基质表现出根据表面结合的肽而变化的蛋白质吸附。与 PEG-g-PA 相比，用线性 PEG 系链连接的肽修饰的 PEG-g-PA 在 24 小时时将蛋白质吸附减少了约 45%。与 PEG-g-PA 相比，通过 StarPEO 和 StarlikePEO 系链连接的肽进一步降低了蛋白质吸附。肽:PEO系链抑制蛋白质吸附的能力似乎是PEO系链的类型和表面覆盖度的函数，并且不受系链肽的量或分子结构的影响。与对照相比，直接偶联至 PEG-g-PA 的肽增加了蛋白质污染量，并且肽所束缚的蛋白质吸附量似乎存在一定依赖性。两种 C-14 标记的病原体，表皮葡萄球菌和铜绿假单胞菌，用于通过两种类型的层流池室来定量细菌粘附程度；一种提供目标基质的侵入性采样，另一种提供粘附细菌细胞的非侵入性显微监测。与碱性聚丙烯酸相比，两种物质对 PEG-g-PA 和肽修饰的 PEG-g-PA 的附着均减少。表面上肽的存在，无论是直接结合还是通过 PEO 系链结合，都不影响铜绿假单胞菌相对于对照的粘附。对于那些肽直接结合到表面的材料，表皮葡萄球菌粘附率略有增加，但当肽通过 PEO 系链结合时，相对于基础 PEG-g-PA 而言，表皮葡萄球菌粘附率降低。所有用单克隆抗体片段修饰的 PEG-g-PA 表面都显着提高了细菌的初始粘附率和最大附着程度。 (C) 2003 Elsevier Ltd. 保留所有权利。
The potential for base poly(ethylene glycol) graft poly(acrylic acid) PEG-g-PA copolymers and surface-modified PEG-g-PA materials to inhibit random protein fouling and bacterial adhesion are investigated. PEG-g-PA co-polymers were synthesized that inhibited non-specific protein and cellular adhesion. PEG-g-PA co-polymers were then covalently modified with either cell adhesion peptides (YRGDS, YEILDV) or fragments of antibodies to monocyte/macrophage integrin receptors (Anti-VLA4, Anti-beta(1), Anti-beta(2), and Anti-CD64) known to enhance macrophage adhesion and, perhaps, modulate their activation. Materials produced in this work were characterized using: hydrophobicity by contact angle; angle-resolved X-ray Photoelectron Spectroscopy to confirm the presence of PEG in the bulk material and the surface; degree of hydration; differential scanning calorimetry; and thermal gravimetric analysis.To evaluate the non-fouling efficacy of the various modified surfaces, three proteins, human serum albumin, human fibronectin (Fraction I) and human immunoglobulin were I-125 labeled. Samples of base PEG-g-PA and PEG-g-PA, modified with various peptides, were exposed to solutions containing either 2 or 200 mug/ml of one of the labeled proteins at 37degreesC for 24 h. PEG-g-PA substrata modified with directly bound peptides exhibited protein adsorption that varied depending upon the surface bounded peptide. PEG-g-PA modified with peptides linked by linear PEG tethers reduced protein adsorption at 24h by similar to45% in comparison to PEG-g-PA. Peptides linked by way of StarPEO and StarlikePEO tethers further decreased protein adsorption in comparison to PEG-g-PA. The ability of peptide:PEOtethers to inhibit protein adsorption appeared to be a function of type and surface coverage of the PEO tether and not influenced by the amount or molecular structure the tethered peptide. Peptides directly coupled to the PEG-g-PA increased the amount of protein fouling relative to controls and there appeared to be some dependency of the amount of protein adsorption on which peptide was tethered.Two C-14-labeled pathogens, Staphylococcus epidermidis and Pseudomonas aeruginosa, were used to quantity the degree of bacterial adhesion using two types of laminar flow cell chambers; one that provided invasive sampling of the target substrata and one that provided non-invasive microscopic surveillance of adhering bacterial cells. Attachment of both species to PEG-g-PA and peptide-modified PEG-g-PA was reduced compared to the basic poly(acrylic acid). Presence of peptides on the surface, whether directly bound or bound by the PEO tether did not influence adhesion of P. aeruginosa relative to controls. S. epidermidis adhesion rates increased slightly for those materials where peptides were directly bound to the surface but were reduced relative to base PEG-g-PA when peptides were bound by PEO tethers. All PEG-g-PA surfaces modified with fragments of monoclonal antibodies dramatically enhanced bacterial initial adhesion rates and maximum extent of attachment. (C) 2003 Elsevier Ltd. All rights reserved.