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A novel whole tooth‐in‐jaw‐bone culture of rat molars: Morphological, immunohistochemical, and laser capture microdissection analysis
Author(s) -
Chokechanachaisakul Uraiwan,
Kaneko Tomoatsu,
Yamanaka Yusuke,
Okiji Takashi,
Suda Hideaki
Publication year - 2012
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.22072
Subject(s) - pulp (tooth) , molar , biology , dentin , immunohistochemistry , ex vivo , organ culture , pathology , laser capture microdissection , in vivo , anatomy , dentistry , medicine , immunology , gene expression , in vitro , paleontology , biochemistry , microbiology and biotechnology , gene
In conventional whole‐tooth culture systems, limitation exists regarding maintenance of the vitality of the dental pulp, because this tissue is encased in rigid dentin walls that hinder nutrition supply. We here report a whole tooth‐in‐jaw‐bone culture system of rat mandibular first molars, where transcardiac perfusion with culture medium was carried out before placement of the jaw bone into culture medium, aiming to facilitate longer time preservation of the dental pulp tissue. Following 7 days of culture, the pulp tissues were analyzed by histology and immunohistochemistry to ED2 (antiresident macrophage). ED2‐positive macrophages were also analyzed for their Class II MHC, interleukin‐6 (IL‐6), and p53 mRNA expression levels by means of immune‐laser capture microdissection (immune‐LCM). Dentin sialophosphoprotein (DSPP) mRNA expression in odontobalstic layer was also examined by LCM. Teeth cultured following saline‐perfusion and nonperfusion served as cultured controls. Normal teeth also served as noncultured controls. Histological examination demonstrated that the structure of the pulp tissue was well preserved in the medium‐perfused explants in contrast to the cultured control groups. The Class II MHC, IL‐6, and p53 mRNA expression levels of ED2‐positive cells and DSPP expression levels of odontoblastic layer tissues in the pulp of medium‐perfused explants were not significantly different from those in the noncultured normal teeth. In conclusion, the structural integrity and mRNA expression in the pulp were maintained at the in vivo level in the ex vivo whole tooth‐in‐jaw‐bone culture system. The system may lay the foundation for studies aiming at defining further histological and molecular mechanism of the pulp. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.