Advertisement

Journal of Bone and Mineral Metabolism

, Volume 37, Issue 3, pp 467–474 | Cite as

FGF2-responsive genes in human dental pulp cells assessed using a rat spinal cord injury model

  • Ken Sugiyama
  • Kosuke Nagashima
  • Takahiro Miwa
  • Yuta Shimizu
  • Tomoko Kawaguchi
  • Kazuki Iida
  • Naritaka Tamaoki
  • Daijiro Hatakeyama
  • Hitomi Aoki
  • Chikara Abe
  • Hironobu Morita
  • Takahiro Kunisada
  • Toshiyuki Shibata
  • Hidefumi Fukumitsu
  • Ken-ichi TezukaEmail author
Original Article

Abstract

The central nervous system in adult mammals does not heal spontaneously after spinal cord injury (SCI). However, SCI treatment has been improved recently following the development of cell transplantation therapy. We recently reported that fibroblast growth factor (FGF) 2-pretreated human dental pulp cells (hDPCs) can improve recovery in a rat model of SCI. This study aimed to investigate mechanisms underlying the curative effect of SCI enhanced via FGF2 pretreatment; we selected three hDPC lines upon screening for the presence of mesenchymal stem cell markers and of their functionality in a rat model of SCI, as assessed using the Basso, Beattie, and Bresnahan score of locomotor functional scale, electrophysiological tests, and morphological analyses. We identified FGF2-responsive genes via gene expression analyses in these lines. FGF2 treatment upregulated GABRB1, MMP1, and DRD2, which suggested to contribute to SCI or central the nervous system. In an expanded screening of additional lines, GABRB1 displayed rather unique and interesting behavior; two lines with the lowest sensitivity of GABRB1 to FGF2 treatment displayed an extremely minor effect in the SCI model. These findings provide insights into the role of FGF2-responsive genes, especially GABRB1, in recovery from SCI, using hDPCs treated with FGF2.

Keywords

Human dental pulp cells Spinal cord injury Fibroblast growth factor 2 GABRB1 

Notes

Acknowledgements

We wish to thank Editage (http://www.editage.jp) for English language editing. This study was supported by the Japan Society for the Promotion of Science (KAKENHI Grant Numbers JP23592697, JP26670799, JP26463001, JP26293426, JP26463000 and JP17H06731).

Additional information

Accession codes: the microarray data were submitted to the NCBI GEO database under the accession number (GSE83902).

Compliance with ethical standards

Ethical approval

All procedures involving human participants were performed in accordance with the ethical standards of the institutional and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. All applicable institutional and national guidelines for the care and use of animals were followed.

Conflict of interest

This manuscript was partly developed with support from Daiichi Sankyo Company Ltd.

Supplementary material

774_2018_954_MOESM1_ESM.tiff (2.1 mb)
Supplementary material 1 (TIFF 2164 kb)
774_2018_954_MOESM2_ESM.tiff (1.3 mb)
Supplementary material 2 (TIFF 1288 kb)
774_2018_954_MOESM3_ESM.tiff (410 kb)
Supplementary material 3 (TIFF 410 kb)
774_2018_954_MOESM4_ESM.tiff (638 kb)
Supplementary material 4 (TIFF 638 kb)
774_2018_954_MOESM5_ESM.tif (20.9 mb)
Supplementary material 5 (TIFF 21361 kb)
774_2018_954_MOESM6_ESM.tif (11.1 mb)
Supplementary material 6 (TIFF 11382 kb)
774_2018_954_MOESM7_ESM.tif (16.4 mb)
Supplementary material 7 (TIFF 16785 kb)
774_2018_954_MOESM8_ESM.pdf (232 kb)
Supplementary material 8 (PDF 233 kb)

References

  1. 1.
    Le Blank K, Frassoni F, Ball L, Locatelli F, Roelofs H, Lewis I, Lanino E, Sundberg B, Bernardo ME, Remberger M, Dini G, Egeler RM, Bacigalupo A, Fibbe W, Ringden O (2008) Mesenchymal stem cells for treatment of steroid-resistant, severe, acute graft-versus-host disease: a phase II study. Lancet 371:1579–1586CrossRefGoogle Scholar
  2. 2.
    Tannna T, Sachan V (2014) Mesenchymal stem cells: potential in treatment of neurodegenerative diseases. Curr Stem Cell Res Ther 9:513–521CrossRefGoogle Scholar
  3. 3.
    Chen C, Hou J (2016) Mesenchymal stem cell-based therapy in kidney transplantation. Stem Cell Res Ther 7:16.  https://doi.org/10.1186/s13287-016-0283-6 CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Chai Y, Jiang X, Ito Y, Bringas P Jr, Han J, Rowitch DH, Soriano P, McMahon AP, Sucov HM (2000) Fate of the mammalian cranial neural crest during tooth and mandibular morphogenesis. Development 127:1671–1679PubMedGoogle Scholar
  5. 5.
    Gronthos S, Brahim J, Li W, Fisher LW, Cherman N, Boyde A, DenBesten P, Robey PG, Shi S (2002) Stem cell properties of human dental pulp stem cells. J Dent Res 81:531–535CrossRefPubMedGoogle Scholar
  6. 6.
    Yamazaki H, Tsuneto M, Yoshino M, Ymamura K, Hayashi S (2007) Potential of dental mesenchymal cells in developing teeth. Stem Cells 25:78–87CrossRefPubMedGoogle Scholar
  7. 7.
    Takeda T, Tezuka Y, Horiuchi M, Hosono K, Iida K, Hatakeyama D, Miyaki S, Kunisada T, Shibata T, Tezuka K (2008) Characterization of dental pulp stem cells of human tooth germs. J Dent Res 87:676–681CrossRefPubMedGoogle Scholar
  8. 8.
    Iida K, Takeda-Kawaguchi T, Tezuka Y, Kunisada T, Shibata T, Tezuka K (2010) Hypoxia enhances colony formation and proliferation but inhibits differentiation of human dental pulp stem cells. Arch Oral Biol 55:648–654CrossRefPubMedGoogle Scholar
  9. 9.
    Iida K, Takeda-Kawaguchi T, Hada M, Yuriguchi M, Aoki H, Tamaoki N, Hatakeyama D, Kunisada T, Shibata T, Tezuka K (2013) Hypoxia-enhanced derivation of iPSCs from human dental pulp cells. J Dent Res 92:905–910CrossRefPubMedGoogle Scholar
  10. 10.
    Takeda-Kawaguchi T, Sugiyama K, Chikusa S, Iida K, Aoki H, Tamaoki N, Hatakeyama D, Kunisada T, Shibata T, Fusaki N, Tezuka K (2014) Derivation of iPSCs after culture of human dental pulp cells under defined conditions. PLoS One 9:e115392CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Tamaoki N, Takahashi K, Tanaka T, Ichisaka T, Aoki H, Takeda-Kawaguchi T, Iida K, Hatakeyama D, Kunisada T, Shibata T, Tezuka K (2010) Dental pulp cells for induced pluripotent stem cell banking. J Dent Res 89:773–778CrossRefPubMedGoogle Scholar
  12. 12.
    Okita K, Matsumura Y, Sato Y, Okada A, Morizane A, Okamoto S, Hong H, Nakagawa M, Tanabe K, Tezuka K, Shibata T, Kunisada T, Takahashi M, Takahashi J, Saji H, Yamanaka S (2011) A more efficient method to generate integration-free human iPS cells. Nat Method 8:409–412CrossRefGoogle Scholar
  13. 13.
    Tamaoki N, Takahashi K, Aoki H, Iida K, Kawaguchi T, Hatakeyama D, Inden M, Chosa N, Ishisaki A, Kunisada T, Shibata T, Goshima N, Yamanaka S, Tezuka K (2014) The homeobox gene DLX4 promotes generation of human induced pluripotent stem cells. Sci Rep 4:7283CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Harrop JS, Hashimoto R, Norvell D, Raich A, Aarabi B, Grossman RG, Guest JD, Tator CH, Chapman J, Fehlings MG (2012) Evaluation of clinical experience using cell-based therapies in patients with spinal cord injury: a systematic review. J Neurosurg Spine 17:230–246CrossRefPubMedGoogle Scholar
  15. 15.
    Sakai K, Yamamoto A, Matsubara K, Nakamura S, Naruse M, Yamagata M, Sakamoto K, Tauchi R, Wakao N, Imagama S, Hibi H, Kadomatsu K, Ishiguro N, Ueda M (2012) Human dental pulp-derived stem cells promote locomotor recovery after complete transection of the rat spinal cord by multiple neuro-regenerative mechanisms. J Clin Investig 122:80–90PubMedGoogle Scholar
  16. 16.
    Matsubara K, Matsushita Y, Sakai K, Kano F, Kondo M, Noda M, Hashimoto N, Imagama S, Ishiguro N, Suzumura A, Ueda M, Furukawa K, Yamamoto A (2015) Secreted ectodomain of sialic acid-binding Ig-like lectin-9 and monocyte chemoattractant protein-1 promote recovery after rat spinal cord injury by altering macrophage polarity. J Neurosci 35:2452–2464CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Nagashima K, Miwa T, Soumiya H, Ushiro D, Takeda-Kawaguchi T, Tamaoki N, Ishiguro S, Sato Y, Miyamoto K, Ohno T, Osawa M, Kunisada T, Shibata T, Tezuka K, Furukawa S, Fukumitsu H (2017) Priming with FGF2 stimulates human dental pulp cells to promote axonal regeneration and locomotor function recovery after spinal cord injury. Sci Rep 7:13500CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Kasai M, Jikoh T, Fukumitsu H, Furukawa S (2014) FGF-2 responsive and spinal cord-resident cells improve locomotor function after spinal cord injury. J Neurotrauma 31:1584–1598CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Lu P, Wang Y, Graham L, McHale K, Gao M, Wu D, Brock J, Blesch A, Rosenzweig ES, Havton LA, Zheng B, Conner JM, Marsala M, Tuszynski MH (2012) Long-distance growth and connectivity of neural stem cells after severe spinal cord injury. Cell 150:1264–1273CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Kunisada T, Tezuka K, Aoki H, Motohashi T (2014) The stemness of neural crest cells and their derivatives. Birth Defects Res C Embryo Today 102:251–262CrossRefGoogle Scholar
  21. 21.
    Wu LW, Wang YL, Christensen JM, Khalifian S, Schneeberger S, Raimondi G, Cooney DS, Lee WP, Brandacher G (2014) Donor age negatively affects the immunoregulatory properties of both adipose and bone marrow derived mesenchymal stem cells. Transpl Immunol 30:122–127CrossRefPubMedGoogle Scholar
  22. 22.
    Nakatsuji N, Nakajima F, Tokunaga K (2008) HLA-haplotype banking and iPS cells. Nat Biotechnol 26:739–740CrossRefGoogle Scholar
  23. 23.
    Xu W, Chi L, Xu R, Ke Y, Luo C, Cai J, Qiu M, Gozal D, Liu R (2005) Increased production of reactive oxygen species contributes to motor neuron death in a compression mouse model of spinal cord injury. Spinal Cord 43:204–213CrossRefPubMedGoogle Scholar
  24. 24.
    Ohashi M, Hirano T, Watanabe K, Shoji H, Ohashi N, Baba H, Endo N, Kohno T (2016) Hydrogen peroxide modulates neuronal excitability and membrane properties in ventral horn neurons of the rat spinal cord. Neuroscience 331:206–220CrossRefPubMedGoogle Scholar
  25. 25.
    Parker DA, Marino V (2013) GABA heteroreceptors modulate noradrenaline release in human dental pulp. J Dent Res 92:1017–1021CrossRefPubMedGoogle Scholar

Copyright information

© The Japanese Society for Bone and Mineral Research and Springer Japan KK, part of Springer Nature 2018

Authors and Affiliations

  • Ken Sugiyama
    • 1
  • Kosuke Nagashima
    • 2
  • Takahiro Miwa
    • 2
  • Yuta Shimizu
    • 3
  • Tomoko Kawaguchi
    • 1
  • Kazuki Iida
    • 1
  • Naritaka Tamaoki
    • 1
  • Daijiro Hatakeyama
    • 1
  • Hitomi Aoki
    • 4
  • Chikara Abe
    • 5
  • Hironobu Morita
    • 5
  • Takahiro Kunisada
    • 4
  • Toshiyuki Shibata
    • 1
  • Hidefumi Fukumitsu
    • 2
  • Ken-ichi Tezuka
    • 4
    • 6
    Email author
  1. 1.Department of Oral and Maxillofacial ScienceGifu University Graduate School of MedicineGifuJapan
  2. 2.Laboratory of Molecular BiologyGifu Pharmaceutical UniversityGifuJapan
  3. 3.Department of PeriodontologyAsahi University School of DentistryMizuhoJapan
  4. 4.Department of Tissue and Organ DevelopmentGifu University Graduate School of MedicineGifuJapan
  5. 5.Department of PhysiologyGifu University Graduate School of MedicineGifuJapan
  6. 6.Center for Highly Advanced Integration of Nano and Life SciencesGifu University (G-CHAIN)GifuJapan

Personalised recommendations