title: The sequential seeding of epithelial and mesenchymal cells for tissue-engineered tooth regeneration.
abstract: Progress is being made toward regenerating teeth by seeding dissociated postnatal odontogenic cells onto scaffolds and implanting them in vivo, but tooth morphology remains difficult to control. In this study, we aimed to facilitate tooth regeneration using a novel technique to sequentially seed epithelial cells and mesenchymal cells so that they formed appropriate interactions in the scaffold. Dental epithelium and mesenchyme from porcine third molar teeth were enzymatically separated and dissociated into single cells. Mesenchymal cells were seeded onto the surface of the scaffold and epithelial cells were then plated on top so that the two cell types were in direct contact. The cell-scaffold constructs were evaluated in vitro and also implanted into immunocompromised rats for in vivo analysis. Control groups included constructs where direct contact between the two cell types was prevented. In scaffolds seed using the novel technique, alkaline phosphatase activity was significantly greater than controls, the tooth morphology in vivo was developed in similar to that of natural tooth, and only one tooth structure formed in each scaffold. These results suggest that the novel cell-seeding technique could be useful for regulating the morphology of regenerated teeth.
authors.affiliation: Tooth Regeneration, Division of Stem Cell Engineering, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan. honda-m@ims.u-tokyo.ac.jp
authors.forename: Masaki J
authors.initials: MJ
authors.lastname: Honda
authors.forename: Shuhei
authors.initials: S
authors.lastname: Tsuchiya
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Sumita
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Sagara
authors.forename: Minoru
authors.initials: M
authors.lastname: Ueda
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D018920:Coculture Techniques; D004847:Epithelial Cells / Q000166:cytology* / Q000201:enzymology / Q000637:transplantation*; D008648:Mesoderm / Q000166:cytology* / Q000201:enzymology / Q000637:transplantation*; D046529:Microscopy, Electron, Transmission; D051381:Rats; D012038:Regeneration; D013552:Swine; D013997:Time Factors; D023822:Tissue Engineering; D014070:Tooth / Q000254:growth & development / Q000502:physiology*
chemical_list: D000469:Alkaline Phosphatase