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. 2018 Apr 25;8:6546. doi: 10.1038/s41598-018-24973-0

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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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Immunohistochemistry staining of AMBN, AMGN, BSP and COL I and Periostin. AMBN, AMGN, BSP and COL I were positively stained at the interfacial layers of iTDM and the tissues opposite to iTDM in CLC groups (indicated by black arrows). These indicated that enamel-like minerals may deposit on surfaces of iTDM seeded with CLC cells. Conversely, HERS cell groups showed negative expression of AMBN and AMGN but positive for BSP, COL I and Periostin. A thin layer at the surface of iTDM in HERS cell groups (indicated by blue arrows) was positively stained for BSP, COL I and Periostin. The fibers attached to the surface layer of the iTDM were also positive for COL I and Periostin. These indicated that cementum and periodontal ligament-like tissues were formed especially in IF_CM and MF_CM-induced HERS cells. Scale bars: 50 μm.