title: Novel biological activity of ameloblastin in enamel matrix derivative.
abstract: OBJECTIVE
  Enamel matrix derivative (EMD) is used clinically to promote periodontal tissue regeneration. However, the effects of EMD on gingival epithelial cells during regeneration of periodontal tissues are unclear. In this in vitro study, we purified ameloblastin from EMD and investigated its biological effects on epithelial cells.
  
  
  METHODS
  Bioactive fractions were purified from EMD by reversed-phase high-performance liquid chromatography using hydrophobic support with a C18 column. The mouse gingival epithelial cell line GE-1 and human oral squamous cell carcinoma line SCC-25 were treated with purified EMD fraction, and cell survival was assessed with a WST-1 assay. To identify the proteins in bioactive fractions of EMD, we used proteome analysis with two-dimensional gel electrophoresis followed by identification with liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis.
  
  
  RESULTS
  Purified fractions from EMD suppressed proliferation of GE-1 and SCC-25. LC-MS/MS revealed that ameloblastin in EMD is the component responsible for inhibiting epithelial cell proliferation. The inhibitory effect of ameloblastin on the proliferation of GE-1 and SCC-25 was confirmed using recombinant protein.
  
  
  CONCLUSIONS
  The inhibitory effects of EMD on epithelial cell proliferation are caused by the biological activities of ameloblastin, which suggests that ameloblastin is involved in regulating epithelial downgrowth in periodontal tissues.
authors.affiliation: Division of Orofacial Functions and Orthodontics, Department of Growth Development of Functions, Kyushu Dental University, Fukuoka, Japan.
authors.forename: Sachiko
authors.initials: S
authors.lastname: Kuramitsu-Fujimoto
authors.affiliation: Division of Infections and Molecular Biology, Department of Health Promotion, Kyushu Dental University, Fukuoka, Japan.
authors.forename: Wataru
authors.initials: W
authors.lastname: Ariyoshi
authors.affiliation: Division of Pulp Biology, Operative Dentistry and Endodontics, Department of Cariology and Periodontology, Kyushu Dental University, Fukuoka, Japan.
authors.forename: Noriko
authors.initials: N
authors.lastname: Saito
authors.affiliation: Division of Infections and Molecular Biology, Department of Health Promotion, Kyushu Dental University, Fukuoka, Japan.
authors.forename: Toshinori
authors.initials: T
authors.lastname: Okinaga
authors.affiliation: Division of Cellular Biosignal Sciences, Department of Biochemistry, Iwate Medical University, Iwate, Japan.
authors.forename: Masaharu
authors.initials: M
authors.lastname: Kamo
authors.affiliation: Division of Cellular Biosignal Sciences, Department of Biochemistry, Iwate Medical University, Iwate, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Ishisaki
authors.affiliation: Department of Oral and Maxillofacial Pathobiology, Institute of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Takata
authors.affiliation: Division of Orofacial Functions and Orthodontics, Department of Growth Development of Functions, Kyushu Dental University, Fukuoka, Japan.
authors.forename: Kazunori
authors.initials: K
authors.lastname: Yamaguchi
authors.affiliation: Division of Infections and Molecular Biology, Department of Health Promotion, Kyushu Dental University, Fukuoka, Japan.
authors.forename: Tatsuji
authors.initials: T
authors.lastname: Nishihara
mesh_terms: D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D003746:Dental Enamel Proteins / Q000494:pharmacology*; D015180:Electrophoresis, Gel, Two-Dimensional; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects*; D005881:Gingiva / Q000166:cytology / Q000187:drug effects; D016557:Guided Tissue Regeneration, Periodontal / Q000379:methods; D006801:Humans; D051379:Mice; D010518:Periodontitis / Q000188:drug therapy; D010519:Periodontium / Q000187:drug effects*; D012016:Reference Values; D015203:Reproducibility of Results; D016622:Silver Staining; D013997:Time Factors
chemical_list: C490889:AMBN protein, human; D003746:Dental Enamel Proteins; C109069:enamel matrix proteins