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Klf10 regulates odontoblast differentiation and mineralization via promoting expression of dentin matrix protein 1 and dentin sialophosphoprotein genes

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Abstract

Klf10, a member of the Krüppel-like family of transcription factors, is critical for osteoblast differentiation, bone formation and mineralization. However, whether Klf10 is involved in odontoblastic differentiation and tooth development has not been determined. In this study, we investigate the expression patterns of Klf10 during murine tooth development in vivo and its role in odontoblastic differentiation in vitro. Klf10 protein was expressed in the enamel organ and the underlying mesenchyme, ameloblasts and odontoblasts at early and later stages of murine molar formation. Furthermore, the expression of Klf10, Dmp1, Dspp and Runx2 was significantly elevated during the process of mouse dental papilla mesenchymal differentiation and mineralization. The overexpression of Klf10 induced dental papilla mesenchymal cell differentiation and mineralization as detected by alkaline phosphatase staining and alizarin red S assay. Klf10 additionally up-regulated the expression of odontoblastic differentiation marker genes Dmp1, Dspp and Runx2 in mouse dental papilla mesenchymal cells. The molecular mechanism of Klf10 in controlling Dmp1 and Dspp expression is thus to activate their regulatory regions in a dosage-dependent manner. Our results suggest that Klf10 is involved in tooth development and promotes odontoblastic differentiation via the up-regulation of Dmp1 and Dspp transcription.

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    Authors and Affiliations

    1. Department of Conservative Dentistry, Affiliated Hospital of Stomatology, Medical College, Zhejiang University, Hangzhou, People’s Republic of China

      Zhuo Chen, Shuli Deng & Hui Chen

    2. Department of Developmental Dentistry, Dental School, The University of Texas Health Science Center at San Antonio, San Antonio, Tex., USA

      Zhuo Chen, Chunyan Wan, Daoshu Luo & Shuo Chen

    3. Department of Pathology, Weifang Medical University, Weifang, Shandong, People’s Republic of China

      Wentong Li

    4. Shangyang Dental Clinic, Hangzhou, People’s Republic of China

      Han Wang

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    1. Zhuo Chen
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    3. Han Wang
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    Correspondence to Hui Chen or Shuo Chen.

    Additional information

    Zhuo Chen and Wentong Li are co-first authors.

    Zhuo Chen and Wentong Li contributed equally to this work.

    This study was supported by grants from the National Natural Science Foundation of China (no. 81100727), the Education Department of Zhejiang Province (no. Y200909390) 2011 China State Key Clinical Department Grants and the National Institute of Dental and Craniofacial Research (NIDCR; RO1 DE019892).

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    Supplemental Fig. 1

    Expression of Klf10 and PCNA during tooth development. Expression pattern of Klf10 and PCNA in developing teeth from embryonic day (E) 12.5 to postnatal day (PN) 20. At E12.5, the expression of PCNA (f) and Klf10 (k) is seen in dental epithelium and the underlying mesenchyme. At E16.5, both PCNA (g) and Klf10 (l) are expressed in the secondary enamel knot, inner and outer enamel epithelia, stratum intermedium and dental mesenchymal cells beneath the inner enamel epithelia. At PN2, positive PCNA (h) and Klf10 (m) staining appears in differentiating ameloblasts and odontoblasts. At PN10 and PN20, PCNA (i, j) and Klf10 (n, o) are expressed in ameloblasts and odontoblasts. a-e Bright-field morphology. p-t Tissue sections were stained with Hoechst to reveal the nucleus. u-y Images were merged (Am ameloblasts, D dentin, de dental epithelia, dp dental pulp, E enamel, od odontoblasts). Bar 50 μM. (GIF 383 kb)

    High resolution image (TIFF 6256 kb)

    Supplemental Fig. 2

    TGF-β1 effect of expression of Klf5, Klf10, Dmp1 and Dspp genes. Mandibles isolated from E14.5 were treated either with or without 10 ng/ml TGF-β1 in DMEM supplemented with 0.5 % FBS, 10 U/ml penicillin and 100 mg/ml streptomycin at 37°C for 2 days. Tissue sections were immunostained with Klf5, Klf10, Dmp1 and Dspp antibodies, respectively. Fluorescent immunohistochemistry showed that TGF-β1 induces expression of Klf5, Klf10, Dmp1 and Dspp genes in dental epithelial and mesenchymal cells (d, j, p, v) compared with that of the control groups (a, g, m, s). b, e, h, k, n, q, t, w Tissue sections were stained with Hoechst to reveal the nucleus. c, f, i, l, o, r, u, x Images were merged. Bar 50 μM. (GIF 375 kb)

    High resolution image (TIFF 5491 kb)

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    Chen, Z., Li, W., Wang, H. et al. Klf10 regulates odontoblast differentiation and mineralization via promoting expression of dentin matrix protein 1 and dentin sialophosphoprotein genes. Cell Tissue Res 363, 385–398 (2016). https://doi.org/10.1007/s00441-015-2260-2

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