Journal of Korean Dental Science

Korean Academy of Dental Science (대한치의학회)
권호 표지
DOI QR코드

DOI QR Code

The Role of Autonomous Wntless in Odontoblastic Differentiation of Mouse Dental Pulp Cells

  • Choi, Hwajung (Cluster for Craniofacial Development and Regeneration Research, Institute of Oral Biosciences, Chonbuk National University School of Dentistry) ;
  • Kim, Tak-Heun (Cluster for Craniofacial Development and Regeneration Research, Institute of Oral Biosciences, Chonbuk National University School of Dentistry) ;
  • Ko, Seung-O (Cluster for Craniofacial Development and Regeneration Research, Institute of Oral Biosciences, Chonbuk National University School of Dentistry) ;
  • Cho, Eui-Sic (Cluster for Craniofacial Development and Regeneration Research, Institute of Oral Biosciences, Chonbuk National University School of Dentistry)
  • Received : 2016.06.09
  • Accepted : 2016.06.22
  • Published : 2016.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

Purpose: Wnt signaling plays an essential role in the dental epithelium and mesenchyme during tooth morphogenesis. Deletion of the Wntless (Wls) gene in odontoblasts appears to reduce canonical Wnt activity, leading to inhibition of odontoblast maturation. However, it remains unclear if autonomous Wnt ligands are necessary for differentiation of dental pulp cells into odontoblast-like cells to induce reparative dentinogenesis, one of well-known feature of pulp repair to form tertiary dentin. Materials and Methods: To analyze the autonomous role of Wls for differentiation of dental pulp cells into odontoblast-like cells, we used primary dental pulp cells from unerupted molars of Wls-floxed allele mouse after infection with adenovirus for Cre recombinase expression to knockout the floxed Wls gene or control GFP expression. The differentiation of dental pulp cells into odontoblast-like cells was analyzed by quantitative real-time polymerase chain reaction. Result: Proliferation rate was significantly decreased in dental pulp cells with Cre expression for Wls knockout. The expression levels of Osterix (Osx), runt-related transcription factor 2 (Runx2), and nuclear factor I-C (Nfic) were all significantly decreased by 0.3-fold, 0.2-fold, and 0.3-fold respectively in dental pulp cells with Wls knockout. In addition, the expression levels of Bsp, Col1a1, Opn, and Alpl were significantly decreased by 0.7-fold, 0.3-fold, 0.8-fold, and 0.6-fold respectively in dental pulp cells with Wls knockout. Conclusion: Wnt ligands produced autonomously are necessary for proper proliferation and odontoblastic differentiation of mouse dental pulp cells toward further tertiary dentinogenesis.

Keywords

References

  1. Goldberg M, Smith AJ. Cells and extracellular matrices of dentin and pulp: a biological basis for repair and tissue engineering. Crit Rev Oral Biol Med. 2004; 15: 13-27. https://doi.org/10.1177/154411130401500103
  2. Mitsiadis TA, Rahiotis C. Parallels between tooth development and repair: conserved molecular mechanisms following carious and dental injury. J Dent Res. 2004; 83: 896-902. https://doi.org/10.1177/154405910408301202
  3. Balic A, Aguila HL, Caimano MJ, Francone VP, Mina M. Characterization of stem and progenitor cells in the dental pulp of erupted and unerupted murine molars. Bone. 2010; 46: 1639-51. https://doi.org/10.1016/j.bone.2010.02.019
  4. Sagomonyants K, Mina M. Biphasic effects of FGF2 on odontoblast differentiation involve changes in the BMP and Wnt signaling pathways. Connect Tissue Res. 2014; 55(Suppl 1): 53-6. https://doi.org/10.3109/03008207.2014.923867
  5. Ricucci D, Loghin S, Lin LM, Spangberg LS, Tay FR. Is hard tissue formation in the dental pulp after the death of the primary odontoblasts a regenerative or a reparative process? J Dent. 2014; 42: 1156-70. https://doi.org/10.1016/j.jdent.2014.06.012
  6. Schroder U. Effects of calcium hydroxidecontaining pulp-capping agents on pulp cell migration, proliferation, and differentiation. J Dent Res. 1985; 64 Spec No: 541-8. https://doi.org/10.1177/002203458506400407
  7. Logan CY, Nusse R. The Wnt signaling pathway in development and disease. Annu Rev Cell Dev Biol. 2004; 20: 781-810. https://doi.org/10.1146/annurev.cellbio.20.010403.113126
  8. Banziger C, Soldini D, Schutt C, Zipperlen P, Hausmann G, Basler K. Wntless, a conserved membrane protein dedicated to the secretion of Wnt proteins from signaling cells. Cell. 2006; 125: 509-22. https://doi.org/10.1016/j.cell.2006.02.049
  9. Fu J, Jiang M, Mirando AJ, Yu HM, Hsu W. Reciprocal regulation of Wnt and Gpr177/mouse Wntless is required for embryonic axis formation. Proc Natl Acad Sci U S A. 2009; 106: 18598-603. https://doi.org/10.1073/pnas.0904894106
  10. Yamashiro T, Zheng L, Shitaku Y, Saito M, Tsubakimoto T, Takada K, Takano-Yamamoto T, ThesleffI. Wnt10a regulates dent in sialophosphoprotein mRNA expression and possibly links odontoblast differentiation and tooth morphogenesis. Differentiation. 2007; 75: 452-62. https://doi.org/10.1111/j.1432-0436.2006.00150.x
  11. Lohi M, Tucker AS, Sharpe PT. Expression of Axin2 indicates a role for canonical Wnt signaling in development of the crown and root during preand postnatal tooth development. Dev Dyn. 2010; 239: 160-7.
  12. Yokose S, Naka T. Lymphocyte enhancer-binding factor 1: an essential factor in odontoblastic differentiation of dental pulp cells enzymatically isolated from rat incisors. J Bone Miner Metab. 2010; 28: 650-8. https://doi.org/10.1007/s00774-010-0185-0
  13. Lin M, Li L, Liu C, Liu H, He F, Yan F, Zhang Y, Chen Y. Wnt5a regulates growth, patterning, and odontoblast differentiation of developing mouse tooth. Dev Dyn. 2011; 240: 432-40. https://doi.org/10.1002/dvdy.22550
  14. Zhu X, Zhao P, Liu Y, Zhang X, Fu J, Ivy Yu HM, Qiu M, Chen Y, Hsu W, Zhang Z. Intra-epithelial requirement of canonical Wnt signaling for tooth morphogenesis. J Biol Chem. 2013; 288: 12080-9. https://doi.org/10.1074/jbc.M113.462473
  15. Kim TH, Lee JY, Baek JA, Lee JC, Yang X, Taketo MM, Jiang R, Cho ES. Constitutive stabilization of ${\beta}$-catenin in the dental mesenchyme leads to excessive dentin and cementum formation. Biochem Biophys Res Commun. 2011; 412: 549-55. https://doi.org/10.1016/j.bbrc.2011.07.116
  16. Kim TH, Bae CH, Lee JC, Ko SO, Yang X, Jiang R, Cho ES. ${\beta}$-catenin is required in odontoblasts for tooth root formation. J Dent Res. 2013; 92: 215-21. https://doi.org/10.1177/0022034512470137
  17. Bae CH, Kim TH, Ko SO, Lee JC, Yang X, Cho ES. Wntless regulates dentin apposition and root elongation in the mandibular molar. J Dent Res. 2015; 94: 439-45. https://doi.org/10.1177/0022034514567198
  18. Balic A, Aguila HL, Mina M. Identification of cells at early and late stages of polarization during odontoblast differentiation using pOBCol3.6GFP and pOBCol2.3GFP transgenic mice. Bone. 2010; 47: 948-58. https://doi.org/10.1016/j.bone.2010.08.009
  19. Choi H, Ahn YH, Kim TH, Bae CH, Lee JC, You HK, Cho ES. TGF-${\beta}$ signaling regulates cementum formation through osterix expression. Sci Rep. 2016; 6: 26046. https://doi.org/10.1038/srep26046
  20. Rathinam E, Rajasekharan S, Chitturi RT, Martens L, De Coster P. Gene expression profiling and molecular signaling of dental pulp cells in response to tricalcium silicate cements: a systematic review. J Endod. 2015; 41: 1805-17. https://doi.org/10.1016/j.joen.2015.07.015
  21. Chen S, Gluhak-Heinrich J, Wang YH, Wu YM, Chuang HH, Chen L, Yuan GH, Dong J, Gay I, MacDougall M. Runx2, osx, and dspp in tooth development. J Dent Res. 2009; 88: 904-9. https://doi.org/10.1177/0022034509342873
  22. Hunter DJ, Bardet C, Mouraret S, Liu B, Singh G, Sadoine J, Dhamdhere G, Smith A, Tran XV, Joy A, Rooker S, Suzuki S, Vuorinen A, Miettinen S, Chaussain C, Helms JA. Wnt acts as a prosurvival signal to enhance dentin regeneration. J Bone Miner Res. 2015; 30: 1150-9. https://doi.org/10.1002/jbmr.2444
  23. Moule AJ, Li H, Bartold PM. Donor variability in the proliferation of human dental pulp fibroblasts. Aust Dent J. 1995; 40: 110-4. https://doi.org/10.1111/j.1834-7819.1995.tb03125.x
  24. Beers MF, Morrisey EE. The three R's of lung health and disease: repair, remodeling, and regeneration. J Clin Invest. 2011; 121: 2065-73. https://doi.org/10.1172/JCI45961
  25. Duan J, Gherghe C, Liu D, Hamlett E, Srikantha L, Rodgers L, Regan JN, Rojas M, Willis M, Leask A, Majesky M, Deb A. Wnt1/${\beta}$-catenin injury response activates the epicardium and cardiac fibroblasts to promote cardiac repair. EMBO J. 2012; 31: 429-42. https://doi.org/10.1038/emboj.2011.418
  26. D'Souza RN, Bachman T, Baumgardner KR, Butler WT, Litz M. Characterization of cellular responses involved in reparative dentinogenesis in rat molars. J Dent Res. 1995; 74: 702-9. https://doi.org/10.1177/00220345950740021301
  27. Smith AJ, Lesot H. Induction and regulation of crown dentinogenesis: embryonic events as a template for dental tissue repair? Crit Rev Oral Biol Med. 2001; 12: 425-37. https://doi.org/10.1177/10454411010120050501