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ABSTRACT: Recently, we demonstrated that a pulse of BrdU given to prenatal animals reveals the existence of slow-cycling long-term label-retaining cells (LRCs), putative adult stem or progenitor cells, which reside in the dental pulp. This study aims to clarify responses of LRCs to allogenic tooth transplantation into mouse maxilla using prenatal BrdU-labeling, in situ hybridization for osteopontin and periostin, and immunohistochemistry for BrdU, nestin, and osteopontin. The upper-right first molars were allografted in the original socket between BrdU-labeled and non-labeled mice or between GFP transgenic and wild-type mice. Tooth transplantation caused degeneration of the odontoblast layer, resulting in the disappearance of nestin-positive reactions in the dental pulp. On postoperative days 5-7, tertiary dentin formation commenced next to the preexisting dentin where nestin-positive odontoblast-like cells were arranged in the successful cases. In BrdU-labeled transplanted teeth, dense LRCs were maintained in the center of the dental pulp beneath the odontoblast-like cells including LRCs, whereas LRCs disappeared in the area surrounding the bone-like tissue. In contrast, LRCs were not recognized in the pulp chamber of non-labeled transplants through the experimental period. Tooth transplantation using GFP mice demonstrated that the donor cells constituted the dental pulp of the transplant except for endothelial cells and some migrated cells, and the periodontal tissue was replaced by host-derived cells except for epithelial cell rests of Malassez. These results suggest that the maintenance of BrdU label-retaining dental pulp cells play a role in the regeneration of odontoblast-like cells in the process of pulpal healing following tooth transplantation.
Histochemie 12/2011; 136(6):649-61. · 2.59 Impact Factor
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ABSTRACT: We have previously demonstrated that tooth size is determined by dental mesenchymal factors. Exogenous bone morphogenetic protein (BMP)4, Noggin, fibroblast growth factor (FGF)3 and FGF10 have no effect on tooth size, despite the expressions of Bmp2, Bmp4, Fgf3, Fgf10 and Lef1 in the dental mesenchyme. Among the wingless (Wnt) genes that are differentially expressed during tooth development, only Wnt5a is expressed in the dental mesenchyme. The aims of the present study were to clarify the expression pattern of Wnt5a in developing tooth germs and the role of Wnt5a in the regulation of tooth size by treatment with exogenous WNT5A with/without an apoptosis inhibitor on in vitro tooth germs combined with transplantation into kidney capsules. Wnt5a was intensely expressed in both the dental epithelium and mesenchyme during embryonic days 14-17, overlapping partly with the expressions of both Shh and Bmp4. Moreover, WNT5A retarded the development of tooth germs by markedly inducing cell death in the non-dental epithelium and mesenchyme but not widely in the dental region, where the epithelial-mesenchymal gene interactions among Wnt5a, Fgf10, Bmp4 and Shh might partly rescue the cells from death in the WNT5A-treated tooth germ. Together, these results indicate that WNT5A-induced cell death inhibited the overall development of the tooth germ, resulting in smaller teeth with blunter cusps after tooth-germ transplantation. Thus, it is suggested that Wnt5a is involved in regulating cell death in non-dental regions, while in the dental region it acts as a regulator of other genes that rescue tooth germs from cell death.
Cell and Tissue Research 09/2011; 345(3):367-77. · 3.11 Impact Factor
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ABSTRACT: We aimed to establish an experimental animal model to evaluate materials for endodontic therapy. We focused on the biocompatibility of new paste-type zinc oxide-eugenol (ZOE) sealer. The results of this sealer were compared with those of conventional powder/liquid ZOE and eugenol-free sealers. The molars of Wistar rats were extracted and repositioned in the original socket after application of the sealers on the root apices. Mild inflammation occurred in the periapical tissue of the replanted teeth with both ZOE sealers on day 7, whereas the eugenol-free sealer induced severe inflammation. On day 14, the lesions induced by all types of sealers were healed and replaced predominantly by fibrous connective tissue. Thus, all endodontic materials showed high biocompatibility, although the extent of inflammatory reactions during the early stages varied depending on the types of materials. We demonstrated that our animal model was useful for the assessment of the biocompatibility of endodontic materials.
Dental Materials Journal 03/2011; 30(2):176-82. · 1.14 Impact Factor
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ABSTRACT: Toll-like receptors (TLRs) are important receptors mediating innate immune responses because they detect factors released from bacterial cell wall components during inflammatory reactions. However, the role of TLRs in dental pulp, which is bounded by hard tissues, is poorly understood. The purpose of this study was to investigate the relationship between the innate immune system and the defense of pulp tissue by using immunodeficient mice that lack an adaptive immune response
Mice with severe combined immunodeficiency (SCID) were used as a model because they lack an adaptive immune response. The expression of TLR-2 and TLR-4 in experimentally inflamed pulps in SCID mice was measured by quantitative real-time polymerase chain reaction and immunohistochemistry. Total RNA was isolated from pulp tissues at 0 to 24 hours after bacterial dentinal infection. Anti-TLR-2, anti-TLR-4 cells, anti-CD64, and antinestin cells were detected with labeled streptavidin-biotin methods.
TLR-2 messenger RNA was detected at 3 hours after bacterial infection and then gradually increased from 9 to 24 hours. Numerous TLR-2- and CD64-positive cells detected on macrophages and dendritic-like cells, and TLR-4- and CD64-positive macrophages were detected in the early stage of pulpitis.
These results suggest that the expression of TLR-2 and TLR-4 may be triggered by bacterial infection in irreversible pulpitis without a need for an adaptive immune response. Those signals may relate to pulpal responses to bacterial infection.
Journal of endodontics 08/2009; 35(7):975-80. · 2.95 Impact Factor
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ABSTRACT: Toll-like receptors (TLRs) are important factors in innate immune responses because they mediate signals from bacterial cell wall components during inflammatory reactions. However, the role of TLR in dental pulp, which is bounded by hard tissues, is little understood. The present study investigated the expression of TLR-2 and TLR-4 in experimentally inflamed pulp by quantitative real-time polymerase chain reaction and immunohistochemistry. Total RNA isolated from pulp tissue from 0 to 72 hours after bacterial dentinal infection. The TLR-2 messenger RNA (mRNA) level was 30-fold higher than the TLR-4 mRNA level at 9 hours. The TLR-2 mRNA level in pulp began to increase by 3 hours after bacterial infection, reaching a maximum level after 9 hours and gradually decreasing from 9 to 72 hours. Numerous TLR-2- and CD64-positive cells detected on macrophage and dendritic-like cells, TLR-4-positive cells detected a little in the pulp at 9 hours. These results suggest that TLR-2 may be mainly regulated during the early stage of pulp inflammation triggered by bacterial infection.
Journal of Endodontics 11/2007; 33(10):1183-6. · 2.88 Impact Factor
