Polystyrene Topography Sticker Array for Cell-Based Assays

Abstract

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.

Surface roughness modulates EGFR signaling and stemness of triple-negative breast cancer cells

“…To investigate the effect of surface microtopography on cell behavior, we first fabricated PS sticker-like topographies with three different levels of roughness generated by microscopic linear scratches characterized by their average surface roughness (Ra). In addition, geometrical micropatterns (depicted by spiral and zigzags) and grooves were generated on the PS using razor printing and polishing methods ( Figure 1A ), as described previously ( Rosado-Galindo and Domenech, 2020 ). Human TNBC cell lines, MDA-MB-231 and MDA-MB-468, were cultured on topographical surfaces, and viability, morphology, and proliferation were measured as initial indicators of potential phenotypic changes in the cells.…”

“…The topographies evaluated in this study were the surface roughness, grooves, and curvature geometries generated on flat polystyrene (PS) films, as described in our previous work ( Stallcop et al, 2018 ; Rosado-Galindo and Domenech, 2020 ). Briefly, a biaxially oriented, 0.19 mm thick PS film (ST311190/3, Goodfellow) and medical-grade tape (ARCare 90106) were used to produce the topographical surfaces.…”

Substrate topographies modulate the secretory activity of human bone marrow mesenchymal stem cells

“…The topographical polystyrene-based (PS-based) array was developed using razor printing, oxygen plasma and sanding methods as described in our previous works (Additional file 1 : Figure S1) [ 29 , 37 ]. Topographies examined were roughness, curvature and wettability on flat PS films.…”

“…Similarly, studies on the effect of spacing, orientation and diameter of micropore/nanorod patterned surfaces proved that straight patterns with relatively low spacing were more effective on MSCs proliferation and differentiation to osteogenic lineage for improved clinical performance of bone implants [ 28 ]. Our previous studies also demonstrated that polystyrene substrates with increased level of roughness enhanced the proliferation of adipose-derived MSCs and the expression of factors associated with proliferation, and thus, this physical stimulation could minimize the need for biochemical supplementation in cultures [ 29 ]. Likewise, surface topography has been shown to influence the behavior of other types of cells.…”

Development of an Optical Method for the Evaluation of Whole Blood Coagulation

“…Methods such as image segmentation allow the identification of structures with specific characteristics [ 25 , 26 ]. To accommodate the blood sample to be processed, simple microfluidic configurations are often used, offering several advantages [ 27 , 28 , 29 , 30 ]. Other techniques, such piezocoagulography, have been also utilized for blood coagulation, showing also the possibility of studying the initial stage of fibrin formation in the clotting process [ 31 ].…”