Cell and Tissue Research

, Volume 356, Issue 2, pp 357–367 | Cite as

Allogenic tooth transplantation inhibits the maintenance of dental pulp stem/progenitor cells in mice

  • Kotaro Saito
  • Mitsushiro Nakatomi
  • Shinichi Kenmotsu
  • Hayato OhshimaEmail author
Regular Article


Our recent study suggested that allogenic tooth transplantation may affect the maintenance of dental pulp stem/progenitor cells. This study aims to elucidate the influence of allograft on the maintenance of dental pulp stem/progenitor cells following tooth replantation and allo- or auto-genic tooth transplantation in mice using BrdU chasing, immunohistochemistry for BrdU, nestin and Ki67, in situ hybridization for Dspp, transmission electron microscopy and TUNEL assay. Following extraction of the maxillary first molar in BrdU-labeled animals, the tooth was immediately repositioned in the original socket, or the roots were resected and immediately allo- or auto-grafted into the sublingual region in non-labeled or the same animals. In the control group, two types of BrdU label-retaining cells (LRCs) were distributed throughout the dental pulp: those with dense or those with granular reaction for BrdU. In the replants and autogenic transplants, dense LRCs remained in the center of dental pulp associating with the perivascular environment throughout the experimental period and possessed a proliferative capacity and maintained the differentiation capacity into the odontoblast-like cells or fibroblasts. In contrast, LRCs disappeared in the center of the pulp tissue by postoperative week 4 in the allografts. The disappearance of LRCs was attributed to the extensive apoptosis occurring significantly in LRCs except for the newly-differentiated odontoblast-like cells even in cases without immunological rejection. The results suggest that the host and recipient interaction in the allografts disturbs the maintenance of dense LRCs, presumably stem/progenitor cells, resulting in the disappearance of these cell types.


Apoptosis Dental pulp Stem cells Transplantation Mice (ICR) 



The authors cordially thank Dr. Helena Ritchie for providing Dspp plasmids. This work was supported by Grants-in-Aid for Scientific Research (B) (no. 22390341 and 25293371 to H.O.) from JSPS and Grant-in-Aid for JSPS Fellows (no. 245934). No potential conflicts of interest have been disclosed.

Supplementary material

Supplementary Data 1

(MPG 21696 kb)

Supplementary Data 2

(MPG 70592 kb)

Supplementary Data 3

(MPG 22080 kb)

441_2014_1818_Fig7_ESM.jpg (1.8 mb)
Supplementary Fig. 1

Nestin-immunoreactivities (a-c) and H&E staining (d) in the allografted (a, d), autografted (b) and replanted teeth (c) at 1 (a-c) and 4 (d) weeks after the operations (D dentin, DP dental pulp). (a-c) Nestin-positive odontoblast-like cells are arranged along the pulp-dentin border. (Inset) higher magnifications of the boxed areas in a, b and c. (d) A sparse connective tissue occupies the dental pulp and odontoblast-like cells disappear from the pulp chamber despite the occurrence of abundant tertiary dentin (TD). Bars 100 μm (a, b, c), 50 μm (d) (JPEG 1842 kb)

441_2014_1818_Fig8_ESM.jpg (2.5 mb)
Supplementary Fig. 2

Nestin- (a), BrdU- (b-e) and Ki-67-immunoreactivities (f) and TUNEL-assay (g) in the control teeth (4 weeks after birth) (D dentin, DP dental pulp, OB odontoblasts). The brown color in the BrdU-immunopositive cells is changed to red. (a) Nestin-positive odontoblasts are arranged along the pulp-dentin border. Inset higher magnification of the boxed area in a. (b) Dental pulp contains numerous label-retaining cells (LRCs) with dense or granular reactions. (c-e) Higher magnification of the boxed areas labeled by c-e in (b), respectively. (c) Two types of LRCs are distinguished: those with dense (arrow) and granular reactions (arrowhead). Dense LRCs are mainly associated with the blood vessels (BV) in the center of the dental pulp, whereas granular LRCs are distributed throughout the dental pulp. (d, e) The coronal odontoblasts include many dense LRCs (arrow), whereas the root odontoblasts lack dense LRCs. (f, g) Ki-67- or TUNEL-positive cells are rarely observed in the control dental pulp. Bars 100 μm (a, b), 50 μm (f, g), 25 μm (c-e) (JPEG 2587 kb)

441_2014_1818_Fig9_ESM.jpg (833 kb)
Supplementary Fig. 3

Ki-67- (a) and PCNA-immunoreactivities (b) in the allografted teeth at 4 weeks after the operation (DP dental pulp). The brown color in the Ki-67- and the PCNA-immunopositive cells is changed to red. (a, b) PCNA-positive reaction is detected more broadly than Ki-67-positive reaction. Bars 50 μm (a, b) (JPEG 833 kb)

441_2014_1818_Fig10_ESM.jpg (2.8 mb)
Supplementary Fig. 4

Semithin sections (a, d, g, j, m, p) and electron micrographs (b, c, e, f, h, i, k, l, n, o, q, r) of the pulp tissues of the allografted (a-c, j-l), autografted (d-f, m-o) and replanted teeth (g-i, p-r) at 1 (a-c) and 2 (d-l) weeks after the operations (D dentin, DP dental pulp, OB odontoblast-like cells, PD predentin). Figures f, i, l, o and r are higher magnifications of the boxed areas in figures e, h, k, n and q, respectively. (a-c) Numerous inflammatory cells such as dendritic cells, macrophages and polymorphonuclear leucocytes (PML) migrate along the dentin-pulp border. (d-r) Newly-differentiated odontoblast-like cells with developed cell organelles such as rER and Golgi apparatus are arranged along the pulp-dentin border. Bars 25 μm (a, d, g, j, m, p), 2 μm (b, c, e, h, k, n, q), 1 μm (f, i, r), 0.5 μm (l, o) (JPEG 2889 kb)


  1. Bergmann A, Steller H (2010) Apoptosis, stem cells, and tissue regeneration. Sci Signal 3:re8PubMedCentralPubMedCrossRefGoogle Scholar
  2. Byers MR, Kvinnsland I, Bothwell M (1992) Analysis of low affinity nerve growth factor receptor during pulpal healing and regeneration of myelinated and unmyelinated axons in replanted teeth. J Comp Neurol 326:470–484PubMedCrossRefGoogle Scholar
  3. Caldwell MA, He X, Svendsen CN (2005) 5-Bromo-2′-deoxyuridine is selectively toxic to neuronal precursors in vitro. Eur J Neurosci 22:2965–2970PubMedCrossRefGoogle Scholar
  4. Hasegawa T, Suzuki H, Yoshie H, Ohshima H (2007) Influence of extended operation time and of occlusal force on determination of pulpal healing pattern in replanted mouse molars. Cell Tissue Res 329:259–272PubMedCrossRefGoogle Scholar
  5. Ishikawa Y, Ida-Yonemochi H, Suzuki H, Nakakura-Ohshima K, Jung HS, Honda MJ, Ishii Y, Watanabe N, Ohshima H (2010) Mapping of BrdU label-retaining dental pulp cells in growing teeth and their regenerative capacity after injuries. Histochem Cell Biol 134:227–241PubMedCrossRefGoogle Scholar
  6. Ishikawa Y, Ida-Yonemochi H, Nakakura-Ohshima K, Ohshima H (2012) The relationship between cell proliferation and differentiation and mapping of putative dental pulp stem/progenitor cells during mouse molar development by chasing BrdU-labeling. Cell Tissue Res 348:95–107PubMedCrossRefGoogle Scholar
  7. Kvinnsland I, Heyeraas KJ, Byers MR (1991) Regeneration of calcitonin gene-related peptide immunoreactive nerves in replanted rat molars and their supporting tissues. Arch Oral Biol 36:815–826PubMedCrossRefGoogle Scholar
  8. Mutoh N, Nakatomi M, Ida-Yonemochi H, Nakagawa E, Tani-Ishii N, Ohshima H (2011) Responses of BrdU label-retaining dental pulp cells to allogenic tooth transplantation into mouse maxilla. Histochem Cell Biol 136:649–661PubMedCrossRefGoogle Scholar
  9. Nakatomi M, Morita I, Eto K, Ota MS (2006) Sonic hedgehog signaling is important in tooth root development. J Dent Res 85:427–431PubMedCrossRefGoogle Scholar
  10. Ogawa R, Saito C, Jung HS, Ohshima H (2006) Capacity of dental pulp differentiation after tooth transplantation. Cell Tissue Res 326:715–724PubMedCrossRefGoogle Scholar
  11. Ohshima H (1990) Ultrastructural changes in odontoblasts and pulp capillaries following cavity preparation in rat molars. Arch Histol Cytol 53:423–438PubMedCrossRefGoogle Scholar
  12. Ohshima H, Nakakura-Ohshima K, Yamamoto H, Maeda T (2001) Alteration in the expression of heat shock protein (Hsp) 25-immunoreactivity in the dental pulp of rat molars following tooth replantation. Arch Histol Cytol 64:425–437PubMedCrossRefGoogle Scholar
  13. Ohta Y, Ichimura K (2000) Proliferation markers, proliferating cell nuclear antigen, Ki67, 5-bromo-2′-deoxyuridine, and cyclin D1 in mouse olfactory epithelium. Ann Otol Rhinol Laryngol 109:1046–1048PubMedGoogle Scholar
  14. Quispe-Salcedo A, Ida-Yonemochi H, Nakatomi M, Ohshima H (2012) Expression patterns of nestin and dentin sialoprotein during dentinogenesis in mice. Biomed Res 33:119–132PubMedCrossRefGoogle Scholar
  15. Ritchie HH, Berry JE, Somerman MJ, Hanks CT, Bronckers AL, Hotton D, Papagerakis P, Berdal A, Butler WT (1997) Dentin sialoprotein (DSP) transcripts: developmentally-sustained expression in odontoblasts and transient expression in pre-ameloblasts. Eur J Oral Sci 105:405–413PubMedCrossRefGoogle Scholar
  16. Rungvechvuttivittaya S, Okiji T, Suda H (1998) Responses of macrophage-associated antigen-expressing cells in the dental pulp of rat molars to experimental tooth replantation. Arch Oral Biol 43:701–710PubMedCrossRefGoogle Scholar
  17. Saito K, Ishikawa Y, Nakakura-Ohshima K, Ida-Yonemochi H, Nakatomi M, Kenmotsu S, Ohshima H (2011) Differentiation capacity of BrdU label-retaining dental pulp cells during pulpal healing following allogenic transplantation in mice. Biomed Res 32:247–257PubMedCrossRefGoogle Scholar
  18. Saito K, Nakatomi M, Ohshima H (2013) Dynamics of bromodeoxyuridine label-retaining dental pulp cells during pulpal healing after cavity preparation in mice. J Endod 39:1250–1255PubMedCrossRefGoogle Scholar
  19. Schaniel C, Moore KA (2009) Genetic models to study quiescent stem cells and their niches. Ann NY Acad Sci 1176:26–35PubMedCrossRefGoogle Scholar
  20. Shimizu A, Nakakura-Ohshima K, Noda T, Maeda T, Ohshima H (2000) Responses of immunocompetent cells in the dental pulp to replantation during the regeneration process in rat molars. Cell Tissue Res 302:221–233PubMedCrossRefGoogle Scholar
  21. Takamori Y, Suzuki H, Nakakura-Ohshima K, Cai J, Cho SW, Jung HS, Ohshima H (2008) Capacity of dental pulp differentiation in mouse molars as demonstrated by allogenic tooth transplantation. J Histochem Cytochem 56:1075–1086PubMedCentralPubMedCrossRefGoogle Scholar
  22. Terling C, Rass A, Mitsiadis TA, Fried K, Lendahl U, Wroblewski J (1995) Expression of the intermediate filament nestin during rodent tooth development. Int J Dev Biol 39:947–956PubMedGoogle Scholar
  23. Tsukamoto-Tanaka H, Ikegame M, Takagi R, Harada H, Ohshima H (2006) Histochemical and immunocytochemical study of hard tissue formation in dental pulp during the healing process in rat molars after tooth replantation. Cell Tissue Res 325:219–229PubMedCrossRefGoogle Scholar
  24. Unno H, Suzuki H, Nakakura-Ohshima K, Jung HS, Ohshima H (2009) Pulpal regeneration following allogenic tooth transplantation into mouse maxilla. Anat Rec 292:570–579CrossRefGoogle Scholar
  25. Wilson A, Laurenti E, Oser G, van der Wath RC, Blanco-Bose W, Jaworski M, Offner S, Dunant CF, Eshkind L, Bockamp E, Lio P, Macdonald HR, Trumpp A (2008) Hematopoietic stem cells reversibly switch from dormancy to self-renewal during homeostasis and repair. Cell 135:1118–1129PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Kotaro Saito
    • 1
    • 2
  • Mitsushiro Nakatomi
    • 1
  • Shinichi Kenmotsu
    • 1
  • Hayato Ohshima
    • 1
    Email author
  1. 1.Division of Anatomy and Cell Biology of the Hard Tissue, Department of Tissue Regeneration and ReconstructionNiigata University Graduate School of Medical and Dental SciencesChuo-kuJapan
  2. 2.Research Fellow of the Japan Society for the Promotion of ScienceTokyoJapan

Personalised recommendations