Polymer hydrogel particles as biocompatible AFM probes to study CD44/hyaluronic acid interactions on cells
Polymer hydrogel particles as biocompatible AFM probes to study CD44/hyaluronic acid interactions on cells
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DOI：
10.1016/j.polymer.2016.02.019
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发表时间：
2016-10
期刊：
影响因子：
4.6
通讯作者：
Steve Martin;Hanqing Wang;Tino Rathke;U. Anderegg;S. Möller;M. Schnabelrauch;T. Pompe;Stephan Schmidt
中科院分区：
文献类型：
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作者：
Steve Martin;Hanqing Wang;Tino Rathke;U. Anderegg;S. Möller;M. Schnabelrauch;T. Pompe;Stephan Schmidt
我们提出了一种新的基于AFM的方法来检测细胞表面的特定相互作用。这种方法的中心思想是模拟配体和受体介导的细胞表面的生理粘附事件。这样的粘附事件通常发生在机械顺应性和水合表面之间。因此，我们合成了软，配体功能化的水凝胶颗粒，并使用它们作为胶体AFM探针（软胶体探针，SCP）。作为一个原理验证的配体/受体系统，我们研究了透明质酸（HA）配体和CD 44细胞受体之间的相互作用。HA/CD 44相互作用涉及多种信号传导过程、伤口愈合和肿瘤进展。除了HA功能化的聚乙二醇为基础的SCP的详细分析，我们表明，这些SCP选择性地结合到细胞与CD 44受体在其表面通过AFM粘附测量。所有阴性对照均显示粘附力显著降低，证明了该方法的可行性。此外，我们分析了CD 44和HA粘附的时间依赖性，这表明配体/受体相互作用的发展相当快。在接触后约10秒达到最大粘附，进一步表明HA/CD 44相互作用可介导细胞粘附的早期阶段。总的来说，这项工作表明，特定的粘附细胞的相互作用，可以分析接近的生物背景下，通过使用基于水凝胶的胶体AFM探针。
We present a new AFM based methodology to detect specific interactions at cell surfaces. The central idea of this approach is to mimic ligand and receptor mediated physiological adhesion events at cell surfaces. Such adhesion events usually occur between mechanically compliant and hydrated surfaces. Consequently, we synthesized soft, ligand functionalized hydrogel particles and used them as colloidal AFM probes (soft colloidal probes, SCPs). As a proof-of-principle ligand/receptor system we studied the interaction between hyaluronic acid (HA) ligands and CD44 cell receptors. HA/CD44 interactions are involved in manifold signaling processes, in wound healing and tumor progression. Besides a detailed analysis of HA functionalized polyethylene glycol based SCPs, we show that these SCPs selectively bind to cells with CD44 receptors at their surface by means of AFM adhesion measurements. All negative controls showed strongly reduced adhesion forces demonstrating the feasibility of the method. Furthermore we analyzed the time dependence of CD44 and HA adhesion which indicated a rather fast development of the ligand/receptor interaction. Maximum adhesion was reached at about 10 s after contact providing further indication that HA/CD44 interactions can mediate early stages of cell adhesion. Overall this work shows that the specific adhesive interactions of cells can be analyzed close to the biological context by using hydrogel based colloidal AFM probes.