Zhang Hongrui, Gu Qi, Wallace Gordon G, Higgins Michael J
ARC Centre of Excellence for Electromaterials Science, Intelligent Polymer Research Institute, AIIM Facility, Innovation Campus, Squires Way, University of Wollongong, Wollongong, New South Wales 2522, Australia.
Biointerphases. 2018 Jun 11;13(4):041004. doi: 10.1116/1.5022713.
Cell adhesion on conducting polymers is important in organic bioelectronics, including applications such as electronically switchable surfaces and electrochemical transistors. There is a fundamental interest in understanding the conducting polymer-cellular interface though as yet no direct measurements to quantify the cell adhesion forces and energies, particularly at the molecular level, have been undertaken. Here, the authors apply electrochemical-single cell force spectroscopy (EC-SCFS) to directly quantify the de-adhesion forces between single L929 fibroblast cells and polypyrrole doped with dodecylbenzene sulfonate (PPy-DBSA) under electrical stimulation. The EC-SCFS reveals single cell de-adhesion forces of 0.65 nN on PPy-DBSA films with adsorbed fibronectin (FN) protein. Blocking experiments by introducing antibodies show that cell de-adhesion is largely due to the binding (∼60% of interactions) of cell-surface α5β1 integrin receptors. Electrochemical oxidation and reduction of PPy-DBSA during initial adsorption of fibronectin cause a significant decrease in the single cell de-adhesion forces to ∼0.4 nN, which is suggested to relate to electrical stimulation effects on reducing FN adsorption on the polymer. In contrast, when electrical stimulation is applied after protein adsorption is established and during the EC-SCFS measurements, the single cell de-adhesion is significantly enhanced on the oxidized polymer compared to the reduced and nonbiased polymer. The study highlights the use of EC-SCFS to directly quantify cell adhesion on electrode surfaces, as well as the ability to probe molecular-level interactions such as integrin receptor-FN complexes with forces of ∼50-100 pN.
在有机生物电子学中,包括电子可切换表面和电化学晶体管等应用,细胞在导电聚合物上的粘附至关重要。尽管目前尚未进行直接测量来量化细胞粘附力和能量,尤其是在分子水平上,但对于理解导电聚合物 - 细胞界面仍有着根本的兴趣。在此,作者应用电化学单细胞力谱(EC - SCFS)直接量化单个L929成纤维细胞与掺杂十二烷基苯磺酸盐的聚吡咯(PPy - DBSA)在电刺激下的脱粘力。EC - SCFS揭示了在吸附有纤连蛋白(FN)的PPy - DBSA薄膜上单个细胞的脱粘力为0.65 nN。通过引入抗体进行的阻断实验表明,细胞脱粘主要归因于细胞表面α5β1整合素受体的结合(约60%的相互作用)。在纤连蛋白初始吸附过程中PPy - DBSA的电化学氧化和还原导致单个细胞脱粘力显著降低至约0.4 nN,这被认为与电刺激对减少聚合物上FN吸附的影响有关。相比之下,当在蛋白质吸附建立后且在EC - SCFS测量期间施加电刺激时,与还原态和无偏置的聚合物相比,氧化态聚合物上的单个细胞脱粘显著增强。该研究突出了EC - SCFS在直接量化电极表面细胞粘附方面的应用,以及探测分子水平相互作用(如整合素受体 - FN复合物)的能力,其作用力约为50 - 100 pN。