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Distal-less homeobox 2 promotes the osteogenic differentiation potential of stem cells from apical papilla

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Abstract

Dental tissue-derived mesenchymal stem cells (MSCs) are a reliable cell source for dental tissue regeneration. However, the molecular mechanisms underlying the directed differentiation of MSCs remain unclear; thus, their use is limited. The histone demethylase, lysine (K)-specific demethylase 4B (KDM4B), plays critical roles in the osteogenic commitment of MSCs by up-regulating distal-less homeobox 2 (DLX2) expression. The DLX2 gene is highly expressed in dental tissue-derived MSCs but the roles of DLX2 in osteogenesis are unclear. Here, we investigate DLX2 function in stem cells from apical papilla (SCAPs). We found that, in vitro, DLX2 expression was up-regulated in SCAPs by adding BMP4 and by inducing osteogenesis. The knock-down of DLX2 in SCAPs decreased alkaline phosphatase (ALP) activity and mineralization. DLX2 depletion affected the mRNA expression of ALP, bone sialoprotein (BSP) and osteocalcin (OCN) and inhibited SCAP osteogenic differentiation in vitro. Over-expression of DLX2 enhanced ALP activity, mineralization and the expression of ALP, BSP and OCN in vitro. In addition, transplant experiments in nude mice confirmed that SCAP osteogenesis was triggered when DLX2 was activated. Furthermore, DLX2 expression led to the expression of the key transcription factor, osterix (OSX) but not to the expression of runt-related transcription factor 2 (RUNX2). Taken together, these results indicate that DLX2 is stimulated by BMP signaling and enhances SCAP osteogenic differentiation by up-regulating OSX. Thus, the activation of DLX2 signaling might improve tissue regeneration mediated by MSCs of dental origin. These results provide insight into the mechanism underlying the directed differentiation of MSCs of dental origin.

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Acknowledgments

This work was supported by the National Sciences Foundation of China (Grant No. 30872895 and 81300841); by a Key Technology Project Grant for Science and Technology, Department of Hunan Province (Grant No. 2008FJ2011); by the Nature Sciences Foundation of Hunan Province (Grant No. S2013J504B); and by a Project Grant for Science and Technology, Department of Hunan Province (Grant No. 2013SK5075).

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The authors declare no potential conflicts of interest.

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Authors and Affiliations

  1. Xiangya Stomatology Hospital, Central South University, Changsha, 410078, China

    Binbin Qu, Ousheng Liu, Xiaodan Fang, Haixia Zhang, Yuehong Wang, Hongzhi Quan, Jie Zhang, Jing Zhou, Jun Zuo, Jianxia Tang & Zhangui Tang

  2. School of Stomatology, Central South University, Changsha, 410078, Hunan, China

    Binbin Qu, Ousheng Liu, Xiaodan Fang, Haixia Zhang, Yuehong Wang, Hongzhi Quan, Jie Zhang, Jing Zhou, Jun Zuo, Jianxia Tang & Zhangui Tang

  3. Department of Oral and Maxilofacial Surgery, Changsha Stomatological Hospital, Changsha, Hunan, China

    Binbin Qu, Jie Zhang & Jing Zhou

  4. Department of Oncology, The Second Xiangya Hospital, Central South University, Changsha, 410011, China

    Haixia Zhang

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  1. Binbin Qu
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  2. Ousheng Liu
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Correspondence to Zhangui Tang.

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Supplementary Fig. 1

DLX2 expression was not changed at 1, 7 and 21 days after culturing SCAPs in osteogenic-inducing medium compared to expression in SCAPs cultured in normal medium. Real-time RT-PCR results show DLX2 mRNA expression levels. GAPDH was used as an internal control. Student’s t-test was performed to determine statistical significance. All error bars represent SD (n = 3). NS no significant difference. Normal normal culture medium; Induced osteogenic-inducing medium. (JPEG 16 kb)

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Qu, B., Liu, O., Fang, X. et al. Distal-less homeobox 2 promotes the osteogenic differentiation potential of stem cells from apical papilla. Cell Tissue Res 357, 133–143 (2014). https://doi.org/10.1007/s00441-014-1833-9

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