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. 2018 Jul 16;8:10690. doi: 10.1038/s41598-018-29112-3

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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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In vitro angiogenesis assay. (A) Representative CD31 immunostaining image after 11-day culture in the presence of 1 μg/ml of dentin matrix components (DMCs) treated with matrix metalloproteinases (MMP)-1, -2 and -3, incubated DMCs without MMP were used, 20 µM/L suramin as a negative control or 2 ng/ml of vascular endothelial growth factor (VEGF) as a positive control (×40). (B) Quantification of tubule formation. One microgram per millilitre of DMCs treated with MMP-1, -2, and -3 significantly promoted angiogenesis, compared with the negative control, DMCs incubated without MMP and MMPs incubated alone (p < 0.05). No significant differences were observed with 1 µg/ml of DMCs treated with the other MMPs, MMPs incubated alone or at lower concentrations (0.01–0.1 µg/ml) of the above-described MMPs. Groups with similar lower-case letters (i.e., a and b) are not significantly different. Data represent three independent experiments.