Abstract
Introduction
The Wnt signaling pathway plays an important role in tissue development by acting
on proliferation, differentiation, and cell fate decisions. Because the role of Wnt6
in tooth development was still unknown, the purpose of this study was to investigate
the role of Wnt6 in tooth morphogenesis and dental tissue mineralization by elucidating
its effect on human dental papilla cells (hDPCs) in vitro.
Methods
Human dental papilla cells were enzymatically separated from tooth germs. Recombinant
adenovirus encoding full-length Wnt6 cDNA was constructed to overexpress Wnt6, and
the biologic effects of Wnt6 on hDPCs were investigated. Wnt6-transduced changes in
hDPC proliferation were examined by means of a 5-bromodeoxyuridine (BrdU) incorporation
assay and cell cycle analysis. Wnt6-transduced changes in hDPC differentiation were
investigated by evaluating alkaline phosphatase (ALPase) activity, by a mineralization
assay, and analysis of mineralization-related gene expression including ALP, type
I collagen (Col I), osteonectin (ON), osteopontin (OPN), bone sialoprotein (BSP),
and dentin matrix protein-1 (DMP-1).
Results
Wnt6 overexpression had no significant effect on the proliferation of hDPCs by BrdU
incorporation assay and flow cytometric analysis. Wnt6 enhanced differentiation of
hDPCs into functional odontoblast-like cells with up-regulated activity of ALPase
and the expression of mineralization-related genes such as ALP, Col I, ON, OPN, BSP,
and DMP-1. Wnt6 overexpression also promoted the mineralization of hDPCs.
Conclusions
Our findings verified that Wnt6 plays an important role in tooth development by promoting
hDPC differentiation, without significant effects on hDPC proliferation.
Key Words
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Article info
Publication history
Published online: October 26, 2009
Identification
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© 2010 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.
