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首页> 外文期刊>Journal of biomedical materials research. Part B, Applied biomaterials. >Monocyte/macrophage interactions with base and linear-and star-like PEG-modified PEG-poly(acrylic acid) co-polymers
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Monocyte/macrophage interactions with base and linear-and star-like PEG-modified PEG-poly(acrylic acid) co-polymers

机译：单核细胞/巨噬细胞与碱以及线性和星形PEG修饰的PEG-聚（丙烯酸）共聚物的相互作用

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Poly(ethylene glycol): poly(acrylate) PEG-g-PA co-polymers were made that inhibited nonspecific protein and cellular adhesion. PEG-g-PA co-polymers were then covalently modified with either cell adhesion peptides of fragments of antibodies to monocyte/macrophage integrin receptors (anti-ELA4, anti-beta_1, anti-beta_2, and anti-CD64) known to enhance macrophage adhesion and, perhaps, modulate their activation. Peptides were either directly conjugated to the base material or linked by way of PEO-star tethers. Fragments of the antibody region containing the antigen-binding site (Fab' fragments) were coupled to other PEG-g-PA samples using the sulhydryl end groups on Fab' fragments to amine-bearing PEO stars. Macrophage adhesion rates, phagocytic response (oxidative burst), and cytokine expression were determined for each PEG-g-PA material. Luminol-enhanced chemiluminescence was used as a semiquantitative indication of monocyte-macrophage phagocytic activation (oxidative burst). Macrophage cytokine expression in response to control, base, and modified materials was determined by ELISAs for TNF-alpha, IL-1 beta, IL-6, and IL-8. Tissue culture poly(styrene) (TCPS)-mediated the greatest number of adherent monocyte/macrophage cells relative to PEG-g-PA materials. Both YRGDS and YEILDV peptides, whether directly or indirectly (via StarPEO) conjugated to PEG-g-PA, increased adhesion versus controls. Fab' fragments of all four antibodies also promoted enhanced adhesion versus controls. Fab'StarPEO materials presented two orders of magnitude fewer ligands per surface unit area than peptide star materials (10~8 vs. 10~(10)), but were able to adhere similar numbers of cells. For surfaces presenting Fab'_(VLA-4) or YEILDV, both of which may both bind to a cell's VLA-4 receptor, the Star: VLA4 surface showed a greater number of adherent monocyte/macrophages. This result suggests that the Fab' had a higher affinity to the cell receptor than a corresponding minimal peptide binding sequence. All materials exhibited low oxidative burst (luminescence counts per minute, LCPM) per cell DNA without the addition of exogenous stimuli (LCPM/DNA < 100). Directly conjugated peptide materials, poly (propylene) (PP), and TCPS showed the lowest levels of LCPM/DNA without the addition of exogenous stimulus (LCPM.DNA < 20). There was no correlation between LCPM/DNA ratios, with and without added LPS stimulus, versus the individual substrates. Monocyte/macrophages adherent to TCPS substrata showed the overall highest stimulatory potential in cytokine expression response to exogenous LPS, followed by PP > PEG-g-PA > StarPEO. Cells adherent to peptide-modified materials and Fab' -modified materials were overall less stimulated. The method of presenting the peptides (i.e., directly or via Star PEO) influenced the level of cytokine secreted by the adherent macrophage.
机译：制备了聚（乙二醇）：抑制非特异性蛋白质和细胞粘附的聚（丙烯酸酯）PEG-g-PA共聚物。然后，用已知能增强巨噬细胞粘附力的单核细胞/巨噬细胞整合素受体（抗ELA4，抗β_1，抗β_2和抗CD64）抗体片段的任一细胞粘附肽对PEG-g-PA共聚物进行共价修饰并可能调节它们的激活。肽直接偶联到基础材料上，或通过PEO-star系链连接。使用Fab'片段上的巯基末端基团将包含抗原结合位点的抗体区域的片段（Fab'片段）与其他PEG-g-PA样品偶联，以形成含胺的PEO星。确定每种PEG-g-PA材料的巨噬细胞粘附率，吞噬反应（氧化爆发）和细胞因子表达。鲁米诺增强的化学发光被用作单核细胞巨噬细胞吞噬激活（氧化爆发）的半定量指标。通过针对TNF-α，IL-1β，IL-6和IL-8的ELISA确定响应于对照，碱基和修饰的材料的巨噬细胞细胞因子表达。相对于PEG-g-PA材料，组织培养聚（苯乙烯）（TCPS）介导的粘附单核细胞/巨噬细胞数量最多。与对照相比，无论是直接还是间接（通过StarPEO）偶联到PEG-g-PA上的YRGDS和YEILDV肽都增加了粘附力。与对照相比，所有四种抗体的Fab'片段也促进了增强的粘附。 Fab'StarPEO材料的单位表面积配体比肽星形材料的配体少两个数量级（10〜8比10〜（10）），但能够粘附相似数量的细胞。对于呈现Fab'_（VLA-4）或YEILDV的表面，两者都可能与细胞的VLA-4受体结合，Star：VLA4表面显示出更多的粘附单核细胞/巨噬细胞。该结果表明，Fab'对细胞受体的亲和力高于相应的最小肽结合序列。所有材料在不添加外源刺激（LCPM / DNA <100）的情况下，每个细胞DNA均显示出低的氧化爆发力（每分钟的发光计数，LCPM）。直接缀合的肽材料，聚丙烯（PP）和TCPS显示最低水平的LCPM / DNA，而没有添加外源刺激（LCPM.DNA <20）。在添加和不添加LPS刺激的情况下，LCPM / DNA比率与单个底物之间没有相关性。附着于TCPS基质的单核细胞/巨噬细胞在对外源LPS的细胞因子表达响应中显示出总体上最高的刺激潜力，其次是PP> PEG-g-PA> StarPEO。粘附于肽修饰的材料和Fab'修饰的材料的细胞总体上受到较少的刺激。呈递肽的方法（即直接或通过Star PEO）影响了粘附巨噬细胞分泌的细胞因子水平。

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