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.
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).
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.
Our findings verified that Wnt6 plays an important role in tooth development by promoting hDPC differentiation, without significant effects on hDPC proliferation.
To read this article in full you will need to make a payment
Purchase one-time access:Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
One-time access price info
- For academic or personal research use, select 'Academic and Personal'
- For corporate R&D use, select 'Corporate R&D Professionals'
Subscribe:Subscribe to Journal of Endodontics
Already a print subscriber? Claim online access
Already an online subscriber? Sign in
Register: Create an account
Institutional Access: Sign in to ScienceDirect
- Signalling networks regulating dental development.Mech Dev. 1997; 67: 111-123
- Expression of wnt signaling pathway genes during tooth development.Mech Dev. 1999; 85: 197-200
- Wnts as ligands: processing, secretion and reception.Oncogene. 2006; 25: 7461-7468
- Essential role of ADAM28 in regulating the proliferation and differentiation of human dental papilla mesenchymal cells (hDPMCs).Histochem Cell Biol. 2008; 130: 1015-1025
- Cell pellets from dental papillae can reexhibit dental morphogenesis and dentinogenesis.Biochem Biophys Res Commun. 2006; 346: 116-124
- WNT10A and WNT6, clustered in human chromosome 2q35 region with head-to-tail manner, are strongly coexpressed in SW480 cells.Biochem Biophys Res Commun. 2001; 283: 798-805
- Transcriptional targeting to anterior pituitary lactotrophic cells using recombinant adenovirus vectors in vitro and in vivo in normal and estrogen/sulpiride-induced hyperplasic anterior pituitaries.Endocrinology. 2000; 141: 3493-3505
- Expression of transgenes in normal and neoplastic anterior pituitary cells using recombinant adenoviruses: long term expression, cell cycle dependency, and effects on hormone secretion.Endocrinology. 1997; 138: 2184-2194
- In vitro differentiation profile of osteoblasts derived from patients with Saethre-Chotzen syndrome.Bone. 2005; 36: 627-634
- Preparation of PLLA membranes with different morphologies for culture of MG-63 cells.Biomaterials. 2004; 25: 4047-4056
- Sequential expression of matrix protein genes in developing rat teeth.Matrix Biol. 1999; 18: 133-143
- Characteristics of periodontal ligament subpopulations obtained by sequential enzymatic digestion of rat molar periodontal ligament.Bone. 2006; 38: 420-426
- Establishment and characterization of a culture system for enzymatically released rat dental pulp cells.Calcif Tissue Int. 2000; 66: 139-144
- Effects of hydrogen peroxide (H2O2) on alkaline phosphatase activity and matrix mineralization of odontoblast and osteoblast cell lines.Cell Biol Toxicol. 2006; 22: 39-46
- Regulation of cementoblast gene expression by inorganic phosphate in vitro.Calcif Tissue Int. 2006; 78: 103-112
- Colocalization of dentin matrix protein 1 and dentin sialoprotein at late stages of rat molar development.Matrix Biol. 2004; 23: 371-379
- The hidden treasure in apical papilla: the potential role in pulp/dentin regeneration and bioRoot engineering.J Endod. 2008; 34: 645-651
- Prospects for tooth regeneration in the 21st century: a perspective.Microsc Res Tech. 2003; 60: 469-479
- Characterization of the apical papilla and its residing stem cells from human immature permanent teeth: a pilot study.J Endod. 2008; 34: 166-171
- Differentiation of odontoblasts in grafted recombinants of murine epithelial root sheath and dental mesenchyme.Arch Oral Biol. 1989; 34: 27-35
- Hormone-responsive cells derived from human dental papilla: characterization in vitro and in vivo in diffusion chambers.Vitro Cell Dev Biol Plant. 1987; 23: 15-20
- Regenerative endodontics: a review of current status and a call for action.J Endod. 2007; 33: 377-390
- Hepatocyte growth factor exerts promoting functions on murine dental papilla cells.J Endod. 2009; 35: 382-388
- Expression of Wnt signaling pathway gene during tooth development.Mech Dev. 1999; 85: 197-200
- Wnt6 expression in epidermis and epithelial tissues during Xenopus organogenesis.Dev Dyn. 2008; 237: 768-779
- Wnt6 regulates the epithelialisation process of the segment plate mesoderm leading to somite formation.Dev Biol. 2004; 271: 198-209
- Expression patterns of Wnt genes during development of an anterior part of the chicken eye.Dev Dyn. 2006; 235: 496-505
- Wnt6 signaling regulates heart muscle development during organogenesis.Dev Biol. 2008; 323: 177-188
- Wnt6 induces the specification and epithelialization of F9 embryonal carcinoma to primitive endoderm.Cell Signal. 2008; 20: 506-517
- Endochondral and intramembranous fetal bone development: osteoblastic cell proliferation, and expression of alkaline phosphatase, m-twist, and histone H4.J Craniofac Genet Dev Biol. 1996; 16: 94-106
- Isolation of cells from ovine fetal long bone and characterization of their osteoblastic activities during in vitro mineralization.Arch Physiol Biochem. 1997; 105: 158-166
- The human type I collagen mutation database.Nucleic Acids Res. 1997; 25: 181-187
- Identification of a novel isoform of mouse dentin matrix protein 1: spatial expression in mineralized tissues.J Bone Miner Res. 1998; 13: 422-431
- Dual functional roles of dentin matrix protein 1: implications in biomineralization and gene transcription by activation of intracellular Ca2+ store.J Biol Chem. 2003; 278: 17500-17508
- In vitro effects of dentin matrix protein-1 on hydroxyapatite formation provide insights into in vivo functions.J Biol Chem. 2004; 279: 18115-18120
Published online: October 26, 2009
© 2010 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.