Scalisi Silvia, Pennacchietti Francesca, Keshavan Sandeep, Derr Nathan D, Diaspro Alberto, Pisignano Dario, Pierzynska-Mach Agnieszka, Dante Silvia, Cella Zanacchi Francesca
Nanoscopy and NIC@IIT, Istituto Italiano di Tecnologia, 16152 Genoa, Italy.
DIFILAB, Department of Physics, University of Genoa, 16146 Genoa, Italy.
Membranes (Basel). 2021 Nov 15;11(11):878. doi: 10.3390/membranes11110878.
Single Layer Graphene (SLG) has emerged as a critically important nanomaterial due to its unique optical and electrical properties and has become a potential candidate for biomedical applications, biosensors, and tissue engineering. Due to its intrinsic 2D nature, SLG is an ideal surface for the development of large-area biosensors and, due to its biocompatibility, can be easily exploited as a substrate for cell growth. The cellular response to SLG has been addressed in different studies with high cellular affinity for graphene often detected. Still, little is known about the molecular mechanism that drives/regulates the cellular adhesion and migration on SLG and SLG-coated interfaces with respect to other substrates Within this scenario, we used quantitative super-resolution microscopy based on single-molecule localization to study the molecular distribution of adhesion proteins at the nanoscale level in cells growing on SLG and glass. In order to reveal the molecular mechanisms underlying the higher affinity of biological samples on SLG, we exploited stochastic optical reconstruction microscopy (STORM) imaging and cluster analysis, quantifying the super-resolution localization of the adhesion protein vinculin in neurons and clearly highlighting substrate-related correlations. Additionally, a comparison with an epithelial cell line (Chinese Hamster Ovary) revealed a cell dependent mechanism of interaction with SLG.
单层石墨烯（SLG）因其独特的光学和电学性质已成为一种至关重要的纳米材料，并已成为生物医学应用、生物传感器和组织工程的潜在候选材料。由于其固有的二维性质，SLG是开发大面积生物传感器的理想表面，并且由于其生物相容性，可以很容易地用作细胞生长的基质。在不同的研究中已经探讨了细胞对SLG的反应，经常检测到细胞对石墨烯具有高亲和力。然而，关于驱动/调节细胞在SLG以及与其他底物相比的SLG涂层界面上的粘附和迁移的分子机制，我们仍然知之甚少。在这种情况下，我们使用基于单分子定位的定量超分辨率显微镜来研究在SLG和玻璃上生长的细胞中粘附蛋白在纳米尺度上的分子分布。为了揭示生物样品对SLG具有更高亲和力的潜在分子机制，我们利用随机光学重建显微镜（STORM）成像和聚类分析，量化了粘附蛋白纽蛋白在神经元中的超分辨率定位，并清楚地突出了与底物相关的相关性。此外，与上皮细胞系（中国仓鼠卵巢细胞）的比较揭示了细胞与SLG相互作用的依赖机制。
