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. 2020 Jul 2;10:10893. doi: 10.1038/s41598-020-67449-w

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© The Author(s) 2020

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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Influence of different cell confluences with and without loading compression in gene expression. (A) Illustration of the loading compression (LC) setup. Cell monolayer loaded with a sterile glass cylinder in 6-well plate to apply a static compression of 2 g/cm². (B) Light microscopy photography of cells before loading (control) and before unloading (LC 6 h and LC 24 h). ×200 magnification, cropped, exposure adjustment and sharpening in Photoshop CC. (C) mRNA expression of selected markers of inflammation, osteoblastic and osteoclastic markers mean ± SD, n = 6 (two independent experiments in triplicate); normalization by ddC_t method to Rpl22 and control 100% statistical analysis with GraphPad Prism 7, two-way-ANOVA, p < 0.05, ^#control vs. LC (loading compression); *60% vs. 100%; °LC 6 h vs. LC 48 h.