Zhang Ying, Liu Xiao, Michelson Kayla, Trivedi Rachana, Wu Xu, Schepp Eric, Xing Yuqian, Darland Diane, Zhao Julia Xiaojun
Shijiazhuang Center for Disease Control and Prevention, Shijiazhuang 050019, P.R. China.
ACS Biomater Sci Eng. 2018 May 14;4(5):1505-1517. doi: 10.1021/acsbiomaterials.8b00190. Epub 2018 Mar 29.
One of the major challenges associated with modeling the influence of the cellular microenvironment on cell growth and differentiation is finding suitable substrates for growing the cells in a manner that recapitulates the cell-cell and cell-microenvironmental interactions in vitro. As one approach to address this challenge, we have developed graphene oxide (GO)-3D mesh with tunable hardness and porosity for application in cell culture systems. The synthetic method of GO-3D mesh is simple, easily reproducible, and low cost. The foundation of the method is the combination of poly(ethylene)(glycol) (PEG) and GO together with a salt-leaching approach (NaCl) in addition to a controlled application of heat during the synthetic process to tailor the mechanical properties, porosity, and pore-size distribution of the resulting GO-3D mesh. With this methodology, the hydrogel formed by PEG and GO generates a microporous mesh in the presence of the NaCl, leading to the formation of a stable 3D scaffold after extensive heating and washing. Varying the ratio of NaCl to GO controls porosity, pore size, and pore connectivity for the GO-3D mesh. When the porosity is less than 90%, with an increasing ratio of NaCl to GO, the number of pores increases with good interconnectivity. The 3D-mesh showed excellent biocompatibility with vascular cells which can take on a morphology comparable to that observed in vessels in vivo. Cell proliferation and gene expression can be determined from cells grown on the GO-3D scaffold, providing a valuable tool for investigating cell-microenvironmental changes. The GO-3D mesh described results from the synergy of the combined chemical properties of the PEG and GO with the salt-leaching methodology to generate a unique and flexible mesh that can be modified and optimized for a variety of in vitro applications.
模拟细胞微环境对细胞生长和分化的影响所面临的主要挑战之一是找到合适的基质,以便以一种能够在体外重现细胞间和细胞与微环境相互作用的方式来培养细胞。作为应对这一挑战的一种方法,我们开发了具有可调硬度和孔隙率的氧化石墨烯(GO)三维网格,用于细胞培养系统。GO三维网格的合成方法简单、易于重现且成本低廉。该方法的基础是将聚(乙二醇)(PEG)和GO与盐析法(NaCl)相结合,此外在合成过程中通过控制加热来调整所得GO三维网格的机械性能、孔隙率和孔径分布。通过这种方法,PEG和GO形成的水凝胶在NaCl存在下会产生微孔网格,经过充分加热和洗涤后形成稳定的三维支架。改变NaCl与GO的比例可控制GO三维网格的孔隙率、孔径和孔隙连通性。当孔隙率小于90%时,随着NaCl与GO比例的增加,孔隙数量增加且连通性良好。这种三维网格对血管细胞表现出优异的生物相容性,血管细胞呈现出与体内血管中观察到的形态相当的形态。可以从在GO三维支架上生长的细胞中测定细胞增殖和基因表达,这为研究细胞与微环境的变化提供了一个有价值的工具。所描述的GO三维网格是PEG和GO的化学性质与盐析方法协同作用产生的结果,从而生成一种独特且灵活的网格,可针对各种体外应用进行修改和优化。