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. 2018 Jan 18;8:1071. doi: 10.1038/s41598-018-19484-x

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© The Author(s) 2018

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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S. epidermidis biofilms grown on different surfaces over a period of 6 days. (a) Representative CLSM images with LIVE/DEAD staining on different surfaces. (b) The images (1, 3, and 5) were taken at magnification of 8000×; Images (2, 4, and 6) were higher magnification (25000×) of the biofilms. More mature biofilms were found on the polished and the spear-type titanium surfaces, with extensive EPS (red arrows). By contrast, the pocket-type surface continued to exhibit small bacterial clusters that tended to wrap (red dash lines) around at the nano-spears that formed the pockets, with relatively little EPS (red arrows). (c) Biomass volume per area on the nanostructured substrates. *Statistically significant difference (p < 0.05). (d) SEM image taken at magnification of 100, 000×, showed that S. epidermidis cells were further damaged and collapsed inside the pockets (red dash line) after 6 days. Three independent experiments were performed for each substrate type.