Rosado-Galindo Heizel, Domenech Maribella
Mayagüez Campus-Bioengineering Program, University of Puerto Rico, Mayagüez, Puerto Rico.
Mayagüez Campus-Department of Chemical Engineering, University of Puerto Rico, Mayagüez, Puerto Rico.
Recent Prog Mater. 2020;2(2). doi: 10.21926/rpm.2002013. Epub 2020 May 14.
Cells can respond to different topographical cues in their natural microenvironment. Hence, scientists have employed microfabrication techniques and materials to generate culture substrates containing topographies for cell-based assays. However, one of the limitations of custom topographical platforms is the lack of adoption by the broad research community. These techniques and materials have high costs, require high technical expertise, and can leach components that may introduce artifacts. In this study, we developed an array of culture surfaces on polystyrene using razor printing and sanding methods to examine the impact of microscale topographies on cell behavior. The proposed technology consists of culture substrates of defined roughness, depth, and curvature on polystyrene films bound to the bottom of a culture well using double-sided medical-grade tape. Human monocytes and adult mesenchymal stem cells (hMSCs) were used as test beds to demonstrate the applicability of the array for cell-based assays. An increase in cell elongation and Arg-1 expression was detected in macrophages cultured in grooves and on rough substrates as compared to flat surfaces. Also, substrates with enhanced roughness stimulated the proliferation of hMSCs. This effect correlated with the secretion of proteins involved in cell proliferation and the downregulation of those associated with cell differentiation. Our results showed that the polystyrene topography sticker array supports cellular changes guided by microscale surface roughness and geometries. Consequently, microscale surface topographies on polished and razor-printed polystyrene films could leverage the endogenous mechanisms of cells to stimulate cellular changes at the functional level for cell-based assays.
细胞能够对其天然微环境中的不同拓扑线索做出反应。因此，科学家们采用微加工技术和材料来制造含有拓扑结构的培养底物，用于基于细胞的分析。然而，定制拓扑平台的局限性之一是缺乏广大研究群体的采用。这些技术和材料成本高昂，需要高技术专业知识，并且可能会渗出可能引入假象的成分。在本研究中，我们使用剃刀打印和打磨方法在聚苯乙烯上开发了一系列培养表面，以研究微观拓扑结构对细胞行为的影响。所提出的技术包括使用双面医用级胶带将聚苯乙烯薄膜上具有确定粗糙度、深度和曲率的培养底物固定在培养孔底部。人类单核细胞和成人间充质干细胞（hMSCs）被用作测试平台，以证明该阵列在基于细胞的分析中的适用性。与平坦表面相比，在凹槽中和粗糙底物上培养的巨噬细胞中检测到细胞伸长和Arg-1表达增加。此外，粗糙度增强的底物刺激了hMSCs的增殖。这种效应与参与细胞增殖的蛋白质分泌以及与细胞分化相关的蛋白质的下调有关。我们的结果表明，聚苯乙烯拓扑贴纸阵列支持由微观表面粗糙度和几何形状引导的细胞变化。因此，抛光和剃刀打印的聚苯乙烯薄膜上的微观表面拓扑结构可以利用细胞的内源性机制，在功能水平上刺激基于细胞的分析中的细胞变化。
