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. 2019 Oct 14;9:14762. doi: 10.1038/s41598-019-51381-9

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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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Odontoblast differentiation of SHED in the response to the sustained HP. (a) Alizarin Red S staining of the mouse dental mesenchymal cell line mDP cultured with induction (differentiation) medium for 10 days. (b) Effects of the sustained hydrostatic pressure (HP) on the differentiation of mDP cells cultured with a different medium height of 5, 10, 15, 20, and 25 cm. Alizarin red S staining was pefomed after 5 days induction. (c) A schematic diagram shows the sustained HP methods using the same amount of medium at different medium height. (d) Alizarin Red S staining of multipotent stem cells from human exfoliated deciduous teeth (SHED) cultured for 7 and 14 days with or without the HP by the medium height of 5 cm (H: 5cm). The data was representative of 3 independent experiments showed similar results. (e) Expression levels of odontogenic marker genes in SHED. Total RNA was extracted after 72 hrs of odontogenic induction with or without the HP by the medium height of 5 cm (H: 5 cm). Gene expression was analyzed by real-time RT-PCR. The data were pooled from three independent experiments. The error bars indicate standard deviations. Statistical analysis was performed using analysis of variance (***p < 0.01).