Department of Materials Science & Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Acta Biomater. 2013 Jul;9(7):7354-61. doi: 10.1016/j.actbio.2013.02.035. Epub 2013 Feb 27.
There is wide anecdotal recognition that biological cell viability and behavior can vary significantly as a function of the source of commercial tissue culture polystyrene (TCPS) culture vessels to which those cells adhere. However, this marked material dependency is typically resolved by selecting and then consistently using the same manufacturer's product - following protocol - rather than by investigating the material properties that may be responsible for such experimental variation. Here, we quantified several physical properties of TCPS surfaces obtained from a wide range of commercial sources and processing steps, through the use of atomic force microscopy (AFM)-based imaging and analysis, goniometry and protein adsorption quantification. We identify qualitative differences in surface features, as well as quantitative differences in surface roughness and wettability that cannot be attributed solely to differences in surface chemistry. We also find significant differences in cell morphology and proliferation among cells cultured on different TCPS surfaces, and resolve a correlation between nanoscale surface roughness and cell proliferation rate for both cell types considered. Interestingly, AFM images of living adherent cells on these nanotextured surfaces demonstrate direct interactions between cellular protrusions and topographically distinct features. These results illustrate and quantify the significant differences in material surface properties among these ubiquitous materials, allowing us to better understand why the dish can make a difference in biological experiments.
人们普遍认为,生物细胞的活力和行为会随着其附着的商用组织培养聚苯乙烯(TCPS)培养器皿的来源而发生显著变化。然而,这种明显的材料依赖性通常是通过选择并始终使用相同制造商的产品来解决的——遵循既定方案,而不是通过研究可能导致这种实验差异的材料特性来解决。在这里,我们通过原子力显微镜(AFM)成像和分析、接触角测量和蛋白质吸附定量等方法,对来自广泛商业来源和不同加工步骤的 TCPS 表面的多种物理特性进行了定量分析。我们发现表面特征存在定性差异,表面粗糙度和润湿性存在定量差异,而这些差异不能仅仅归因于表面化学性质的差异。我们还发现,在不同的 TCPS 表面培养的细胞形态和增殖存在显著差异,并确定了两种细胞类型的纳米级表面粗糙度与细胞增殖率之间存在相关性。有趣的是,在这些纳米结构表面上培养的活贴壁细胞的 AFM 图像显示了细胞突起与形貌上明显不同的特征之间的直接相互作用。这些结果说明了并量化了这些常见材料之间的材料表面特性的显著差异,使我们能够更好地理解为什么培养皿在生物实验中会产生影响。