title: Evaluation of a dental pulp-derived cell sheet cultured on amniotic membrane substrate.
abstract: OBJECTIVE
  Mesenchymal stem cells (MSC) are transplanted for periodontal tissue regeneration, and the periodontal ligament (PDL) is regenerated using a cultured cell sheet. This cultured cell sheet is prepared using PDL-derived cells, growth factors, and amniotic membrane (AM). Dental pulp (DP)-derived cells can be easily obtained from extracted wisdom teeth, proliferate rapidly, and are less susceptible to bacterial infection than PDL-derived cells. Thus, to prepare a novel cell sheet, DP-derived cells were cultured on AM as a culture substrate for immunohistochemical examination.
  
  
  METHODS
  Wisdom teeth extracted from three adults were cut along the cement-enamel border. DP tissue was collected, minced, and primarily cultured. After three or four passage cultures, DP-derived cells were cultured on AM, followed by hematoxylin-eosin (H-E) and immunofluorescence staining.
  
  
  RESULTS
  DP-derived cells cultured on AM formed a layered structure. Cells positive for vimentin, Ki-67, ZO-1, desmoplakin, CD29, 44, 105 or 146, STRO-1, collagen IV or VII or laminin 5 or α5 chain were localized.
  
  
  CONCLUSIONS
  DP-derived cells proliferated on AM, while retaining the properties of DP, which allowed the cultured cell sheet to be prepared. In addition, the cultured cell sheet contained MSC, which suggests its potential application in periodontal tissue regeneration.
authors: Department of Dental Medicine, Kyoto Prefectural University of Medicine, Graduate School of Medical Science, Kyoto, Japan Department of Immunology, Kyoto Prefectural University of Medicine, Graduate School of Medical Science, Kyoto, Japan.
authors: Ken-ichi
authors: K
authors: Honjo
authors: Department of Dental Medicine, Kyoto Prefectural University of Medicine, Graduate School of Medical Science, Kyoto, Japan.
authors: Toshiro
authors: T
authors: Yamamoto
authors: Department of Dental Medicine, Kyoto Prefectural University of Medicine, Graduate School of Medical Science, Kyoto, Japan.
authors: Tetsuya
authors: T
authors: Adachi
authors: Department of Dental Medicine, Kyoto Prefectural University of Medicine, Graduate School of Medical Science, Kyoto, Japan.
authors: Takeshi
authors: T
authors: Amemiya
authors: Department of Immunology, Kyoto Prefectural University of Medicine, Graduate School of Medical Science, Kyoto, Japan.
authors: Osam
authors: O
authors: Mazda
authors: Department of Dental Medicine, Kyoto Prefectural University of Medicine, Graduate School of Medical Science, Kyoto, Japan.
authors: Narisato
authors: N
authors: Kanamura
authors: Department of Immunology, Kyoto Prefectural University of Medicine, Graduate School of Medical Science, Kyoto, Japan.
authors: Masakazu
authors: M
authors: Kita
mesh_terms: D000328:Adult; D000650:Amnion / Q000737:chemistry*; D049109:Cell Proliferation; D002478:Cells, Cultured; D003782:Dental Pulp / Q000166:cytology*; D051180:Desmoplakins / Q000235:genetics / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism; D005260:Female; D005819:Genetic Markers; D006801:Humans; D007150:Immunohistochemistry; D019394:Ki-67 Antigen / Q000235:genetics / Q000378:metabolism; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D010513:Periodontal Ligament / Q000166:cytology; D023822:Tissue Engineering; D014746:Vimentin / Q000235:genetics / Q000378:metabolism; D014945:Wound Healing; D055815:Young Adult; D062826:Zonula Occludens-1 Protein / Q000235:genetics / Q000378:metabolism
chemical_list: D051180:Desmoplakins; D005819:Genetic Markers; D019394:Ki-67 Antigen; C569042:TJP1 protein, human; D014746:Vimentin; D062826:Zonula Occludens-1 Protein
keywords: Amniotic membrane; dental pulp-derived cells; immunohistochemistry