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使用具有耗散功能的石英晶体微天平监测钽和氧化聚苯乙烯上的细胞粘附。

Monitoring cell adhesion on tantalum and oxidised polystyrene using a quartz crystal microbalance with dissipation.

作者信息

Lord Megan Susan, Modin Charlotte, Foss Morten, Duch Mogens, Simmons Anne, Pedersen Finn S, Milthorpe Bruce K, Besenbacher Flemming

机构信息

Graduate School of Biomedical Engineering, The University of New South Wales, Sydney 2052, Australia.

出版信息

Biomaterials. 2006 Sep;27(26):4529-37. doi: 10.1016/j.biomaterials.2006.04.006. Epub 2006 May 22.

Abstract

The quartz crystal microbalance with dissipation (QCM-D) (Q-Sense AB, Sweden) has been established as a useful tool for evaluating interactions between various biological and non-biological systems, and there has been increasing interest in using the QCM-D technique for cell monitoring applications. This study investigated the potential of the QCM-D to characterise the initial adhesion and spreading of cells in contact with protein precoated biocompatible surfaces. The QCM-D technique is attractive for monitoring cell adhesion and spreading as it allows in situ real-time measurements. The adhesion of NIH3T3 (EGFP) fibroblasts to tantalum (Ta) and oxidised polystyrene (PS(ox)) surfaces precoated with serum proteins was examined using the QCM-D for a period of either 2 or 4 h. Time-lapse photography was performed at 30 min intervals to visually examine cell adhesion and spreading in order to relate cell morphology to the QCM-D response. Following adsorption of albumin, fibronectin or newborn calf serum onto the surfaces, QCM-D measurements showed that cells adhered and spread on the fibronectin and serum coated surfaces, while few cells adhered to the albumin coated surfaces. Cells adhered to albumin coated surfaces had a rounded morphology. The responses to fibronectin and serum precoated surfaces were quite different for each of the underlying substrates indicating that the process of cell adhesion and spreading elicits different responses depending on both the protein coating composition and the influence of the underlying substrate. The different response may be due to extracellular matrix remodelling as well as cytoskeletal changes. Frequency (f) and dissipation (D) changes associated with cell adhesion were less than would be expected from the Sauerbrey relation due to the viscoelastic properties of the cells.

摘要

石英晶体微天平耗散技术(QCM-D)(瑞典Q-Sense AB公司)已成为评估各种生物和非生物系统之间相互作用的有用工具,并且人们越来越有兴趣将QCM-D技术用于细胞监测应用。本研究调查了QCM-D在表征细胞与蛋白质预涂覆生物相容性表面接触时的初始粘附和铺展方面的潜力。QCM-D技术对于监测细胞粘附和铺展具有吸引力,因为它允许进行原位实时测量。使用QCM-D在2或4小时的时间段内检测了NIH3T3(EGFP)成纤维细胞对预涂有血清蛋白的钽(Ta)和氧化聚苯乙烯(PS(ox))表面的粘附。每隔30分钟进行一次延时摄影,以目视检查细胞粘附和铺展情况,以便将细胞形态与QCM-D响应联系起来。在白蛋白、纤连蛋白或新生小牛血清吸附到表面后,QCM-D测量表明细胞在纤连蛋白和血清包被的表面上粘附并铺展,而很少有细胞粘附在白蛋白包被的表面上。粘附在白蛋白包被表面上的细胞呈圆形形态。对于每种底层底物,对纤连蛋白和血清预涂表面的响应有很大不同,这表明细胞粘附和铺展过程会根据蛋白质涂层组成以及底层底物的影响引发不同的响应。这种不同的响应可能是由于细胞外基质重塑以及细胞骨架变化。由于细胞的粘弹性,与细胞粘附相关的频率(f)和耗散(D)变化小于根据绍布雷关系预期的值。

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