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 Ohshima
Regular Article

Abstract

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.

Keywords

Apoptosis Dental pulp Stem cells Transplantation Mice (ICR) 

Supplementary material

Supplementary Data 1

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Supplementary Data 2

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Supplementary Data 3

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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)

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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)

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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)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Kotaro Saito
    • 1
    • 2
  • Mitsushiro Nakatomi
    • 1
  • Shinichi Kenmotsu
    • 1
  • Hayato Ohshima
    • 1
  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

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