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J Appl Oral Sci. 2015 Jan-Feb;23(1):49-55. doi: 10.1590/1678-775720140291.

Novel biological activity of ameloblastin in enamel matrix derivative.

Author information

1
Division of Orofacial Functions and Orthodontics, Department of Growth Development of Functions, Kyushu Dental University, Fukuoka, Japan.
2
Division of Infections and Molecular Biology, Department of Health Promotion, Kyushu Dental University, Fukuoka, Japan.
3
Division of Pulp Biology, Operative Dentistry and Endodontics, Department of Cariology and Periodontology, Kyushu Dental University, Fukuoka, Japan.
4
Division of Cellular Biosignal Sciences, Department of Biochemistry, Iwate Medical University, Iwate, Japan.
5
Department of Oral and Maxillofacial Pathobiology, Institute of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.

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.

MATERIAL AND 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.

CONCLUSION:

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.

PMID:
25760267
PMCID:
PMC4349119
DOI:
10.1590/1678-775720140291
[Indexed for MEDLINE]
Free PMC Article

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