Hasan Abshar, Pattanayek Sudip K, Pandey Lalit M
Bio-Interface & Environmental Engineering Lab, Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Assam 781039, India.
Macromolecules and Interfaces Laboratory, Department of Chemical Engineering, Indian Institute of Technology Delhi, Hauz Khas, New Delhi 110 016, India.
ACS Biomater Sci Eng. 2018 Sep 10;4(9):3224-3233. doi: 10.1021/acsbiomaterials.8b00795. Epub 2018 Aug 22.
Surface modification plays a vital role in regulating protein adsorption and subsequently cell adhesion. In the present work, we prepared nanoscaled modified surfaces using silanization and characterized them using Fourier-transform infrared spectroscopy (FTIR), water contact angle (WCA), and atomic force microscopy (AFM). Five different (amine, octyl, mixed, hybrid, and COOH) surfaces were prepared based on their functionality and varying wettability and their effect on protein adsorption and initial cell adhesion was investigated. AFM analysis revealed nanoscale roughness on all modified surfaces. Fetal bovine serum (FBS) was used for protein adsorption experiment and effect of FBS was analyzed on initial cell adhesion kinetics (up to 6 h) under three different experimental conditions: (a) with FBS in media, (b) with preadsorbed FBS on surfaces, and (c) incomplete media, i.e., without FBS. Various cell features such as cell morphology/circularity, cell area and nuclei size were also studied for the above stated conditions at different time intervals. The cell adhesion rate as well as cell spread area were highest in the case of surfaces with preadsorbed FBS. We observed higher surface coverage rate by adhering cells on hybrid (rate, 0.073 h) and amine (0.072 h) surfaces followed by COOH (0.062 h) and other surfaces under preadsorbed FBS condition. Surface treated with cells in incomplete media exhibited least adhesion rate, poor cell spreading and improper morphology. Furthermore, we found that initial cell adhesion rate and Δdhered cells (%) linearly increased with the change in α-helix content of adsorbed FBS on surfaces. Among all the modified surfaces and under all three experimental conditions, hybrid surface exhibited excellent properties for supporting cell adhesion and growth and hence can be potentially used as surface modifiers in biomedical applications to design biocompatible surfaces.
表面改性在调节蛋白质吸附以及随后的细胞黏附中起着至关重要的作用。在本研究中，我们使用硅烷化制备了纳米级改性表面，并通过傅里叶变换红外光谱（FTIR）、水接触角（WCA）和原子力显微镜（AFM）对其进行了表征。基于其功能和不同的润湿性制备了五种不同的（胺基、辛基、混合、杂化和羧基）表面，并研究了它们对蛋白质吸附和初始细胞黏附的影响。AFM分析揭示了所有改性表面的纳米级粗糙度。使用胎牛血清（FBS）进行蛋白质吸附实验，并在三种不同的实验条件下分析FBS对初始细胞黏附动力学（长达6小时）的影响：（a）培养基中含有FBS，（b）表面预吸附有FBS，以及（c）不完全培养基，即不含FBS。还在不同时间间隔对上述条件下的各种细胞特征，如细胞形态/圆度、细胞面积和细胞核大小进行了研究。在预吸附有FBS的表面情况下，细胞黏附率和细胞铺展面积最高。我们观察到，在预吸附FBS条件下，杂化（速率为0.073 h）和胺基（0.072 h）表面上黏附的细胞具有更高的表面覆盖率，其次是羧基（0.062 h）和其他表面。在不完全培养基中用细胞处理的表面表现出最低的黏附率、较差的细胞铺展和不合适的形态。此外，我们发现初始细胞黏附率和Δd黏附细胞（%）随着表面吸附的FBS的α-螺旋含量的变化而线性增加。在所有改性表面和所有三种实验条件下，杂化表面表现出优异的支持细胞黏附和生长的性能，因此有可能在生物医学应用中用作表面改性剂来设计生物相容性表面。
