title: A novel whole tooth-in-jaw-bone culture of rat molars: morphological, immunohistochemical, and laser capture microdissection analysis.
abstract: 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.
authors.affiliation: Department of Pulp Biology and Endodontics, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Uraiwan
authors.initials: U
authors.lastname: Chokechanachaisakul
authors.forename: Tomoatsu
authors.initials: T
authors.lastname: Kaneko
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Yamanaka
authors.forename: Takashi
authors.initials: T
authors.lastname: Okiji
authors.forename: Hideaki
authors.initials: H
authors.lastname: Suda
mesh_terms: D000818:Animals; D003470:Culture Media / Q000737:chemistry; D003782:Dental Pulp / Q000473:pathology / Q000502:physiology; D020869:Gene Expression Profiling; D007150:Immunohistochemistry; D007568:Jaw / Q000502:physiology*; D061346:Laser Capture Microdissection; D008963:Molar / Q000033:anatomy & histology / Q000502:physiology*; D009924:Organ Culture Techniques / Q000379:methods; D051381:Rats
chemical_list: D003470:Culture Media