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. 2019 Aug 19;9:12076. doi: 10.1038/s41598-019-48369-w

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© The Author(s) 2019

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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Three stimulation runs, each containing two ES groups and a control, were performed to analyse the effect of different treatment settings. Using 4 V/cm, 6 ms pulse duration and 2 Hz a 1.85 times increase in VEGF concentration was found compared to non-stimulated control. Elevated levels of VEGF were also found when we increased/decreased voltage. In the elevated pulse duration and frequency groups, cells were substantially detached which was in part reflected by increased cell death, especially in the 25 Hz group, explaining the highly increased VEGF A expression. Values are expressed as the mean VEGF expression relative to control. N = 3 per stimulation protocol.