微图案高分子生物材料对蛋白质的竞争性吸附,以及定制的细胞外基质的粘弹性

Alexander Welle , Antonio Chiumiento , Rolando Barbucci
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引用次数: 16

摘要

细胞在生物材料表面的粘附和依赖于锚定的细胞的活力受到有意或自发形成的吸附层的几个参数的影响。细胞/生物材料接触之前和过程中的表面预处理和几个调理步骤影响细胞和生物材料之间界面层的组成、取向、数量和粘弹性。这项工作是为了阐明细胞对两种基于蛋白质或碳水化合物吸附的改性生物材料表面的反应:(a)屏蔽紫外线照射为获得控制血清蛋白吸附和细胞粘附的化学图案底物开辟了一条简单的途径。实现亚细胞大小的结构和产生固定梯度是可能的。为了检测沉积在这些基质上的蛋白质基质,我们应用了石英微平衡技术(QCM-D),该技术除了能够提取血浆蛋白质沉积过程中的质量摄取外,还能够提取粘弹性数据。研究发现,紫外照射对表面进行修饰(图案化)后,表面结合白蛋白的数量和粘度降低。(b)另一种组织工程技术,使用固定的、修饰的和/或交联的透明质酸(HA),一种重要的细胞外基质成分,也通过QCM-D进行了检验。我们的数据表明,HA可以通过碳二亚胺活化,然后应用α,ω-双氨基聚乙二醇来修饰。QCM-D数据可以解释为HA层的硬化和水化水的释放。进一步研究了表面结合超薄透明质酸水凝胶的水化状态和粘弹性行为。通过QCM-D对ECM薄膜的粘弹性参数进行量化,对于解释硬致性,描述机械衬底参数对细胞粘附和运动的影响是有价值的。
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Competitive protein adsorption on micro patterned polymeric biomaterials, and viscoelastic properties of tailor made extracellular matrices

Cell adhesion on biomaterial surfaces and the vitality of anchorage dependent cells is affected by several parameters of an adsorbate layer which is intentionally or spontaneously formed. Surface pre-treatments and several conditioning steps prior and during to the cell/biomaterial contact affect the composition, orientation, quantity and viscoelasticity of the interfacing layer between cells and biomaterial. This work was performed to elucidate the response of cells on two modified biomaterial surfaces based on protein or carbohydrate adsorbates:

  • (a)

    Masked UV irradiations opened a simple route to obtain chemically patterned substrates controlling serum protein adsorption and cell adhesion. It is possible to achieve structures of subcellular size and to produce immobilized gradients. In order to examine the protein matrix deposited on these substrates we applied a quartz microbalance technique (QCM-D) capable to extract viscoelastic data in addition to the mass uptake during plasma protein deposition. It was found that the quantity and viscosity of surface bound albumin is lowered when the surface is modified (patterned) by UV exposure. Hence, the UV modification promotes the competitive adsorption of cell adhesion proteins from the media or upon secretion by the cells and yields to the observed cell patterns.

  • (b)

    Another tissue engineering technique, using immobilized, modified and/or cross linked hyaluronic acid (HA), an important extra cellular matrix component in vivo, is also examined by QCM-D. Our data demonstrate that HA can be modified by an activation with a carbodiimide, followed by the application of an α,ω-bisamino polyethyleneglycol. The QCM-D data can be interpreted as a stiffening of the HA layer combined with the release of hydration water. Further, the hydration state and the viscoelastic behaviour of surface bound ultrathin HA hydrogels was examined.

Quantification of viscoelastic parameters of thin films of ECM by QCM-D is valuable for the interpretation of durotaxis, describing effects of mechanical substrate parameters on the adhesion and motility of cells.

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