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. 2021 Apr 14;11:8168. doi: 10.1038/s41598-021-87658-1

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Smaller topography combined with collagen I coating induced higher cellular FA. MCEC were seeded on uncoated (A) or collagen I coated (B) glass, flat, or 800 nm silk films. After 5 days in culture, cells were stained for nuclei (blue-DAPI), vinculin (green- Oregon Green 488) and actin (red- Alexa Fluor 568) expression. Each vinculin punctate areas were analyzed to determine the FA size, FA area and density. (C) Individual FA size was defined by discrete areas of vinculin measurements and it was significantly larger on silk surfaces and addition of collagen I increased the cell FA size on each substrate. (D) The effect of collagen I was significantly evident when vinculin FA area was calculated, no major differences were observed on uncoated surfaces, but its addition to the 800 nm patterned silk films greatly increased the vinculin image area. (E) FA density was higher in cell cultured on glass across all uncoated substrate. However, addition of collagen I changed the vinculin density, especially on the 800 nm topography (* indicates p < 0.05, ** indicates p < 0.005, ***indicates p < 0.0005. Values are means ± S.E.M.).