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Basic Research| Volume 38, ISSUE 6, P796-802, June 2012

Changes in Proliferation and Osteogenic Differentiation of Stem Cells from Deep Caries In Vitro

Published:March 26, 2012DOI:https://doi.org/10.1016/j.joen.2012.02.014

      Abstract

      Introduction

      It is suggested that dental pulp stem cells are involved in tooth regeneration and play an important role in maintaining pulp homeostasis. Previously, normal dental pulps were more widely used for experimental models than carious dental pulps. The aim of this study was to isolate and culture the dental pulp stem cells from carious and normal teeth and to evaluate stem cell parameters.

      Methods

      Pulp tissues were obtained and dissociated from normal and carious teeth. Single-cell suspensions were seeded into 6-well plates and purified by collecting multiple colonies. Normal dental pulp stem cells (DPSCs) and carious dental pulp stem cells (CDPSCs) were compared for morphologic appearance and for their capacity to differentiate into 3 lineages. Colony-forming and MTT assays, cell cycle analysis, gene expression, and alkaline phosphatase activity were also evaluated.

      Results

      Stem cells were cultured successfully from normal and carious dental pulps. CDPSCs displayed increased proliferation ability compared with DPSCs. CDPSCs also showed enhanced ALP activity, mineralization ability, and expression of osteogenesis/dentinogenesis-related genes. All cultures differentiated into 3 cell types.

      Conclusions

      Our data suggest that caries as a local microenvironment should be taken into account when DPSCs are intended to be used for investigations and application. Furthermore, the mechanism of the underlying changes in cell properties requires further study.

      Key Words

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      References

        • Gronthos S.
        • Mankani M.
        • Brahim J.
        • Robey P.G.
        • Shi S.
        Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo.
        Proc Natl Acad Sci U S A. 2000; 97: 13625-13630
        • Koyama N.
        • Okubo Y.
        • Nakao K.
        • Bessho K.
        Evaluation of pluripotency in human dental pulp cells.
        J Oral Maxillofac Surg. 2009; 67: 501-506
        • Yu J.
        • He H.
        • Tang C.
        • et al.
        Differentiation potential of STRO-1+ dental pulp stem cells changes during cell passaging.
        BMC Cell Biol. 2010; 11: 32
        • Graziano A.
        • d’Aquino R.
        • Laino G.
        • Papaccio G.
        Dental pulp stem cells: a promising tool for bone regeneration.
        Stem Cell Rev. 2008; 4: 21-26
        • d’Aquino R.
        • Graziano A.
        • Sampaolesi M.
        • et al.
        Human postnatal dental pulp cells co-differentiate into osteoblasts and endotheliocytes: a pivotal synergy leading to adult bone tissue formation.
        Cell Death Differ. 2007; 14: 1162-1171
        • Tziafas D.
        • Kodonas K.
        Differentiation potential of dental papilla, dental pulp, and apical papilla progenitor cells.
        J Endod. 2010; 36: 781-789
        • Marigo L.
        • Migliaccio S.
        • Monego G.
        • La Torre G.
        • Somma F.
        • Ranelletti F.O.
        Expression of parathyroid hormone-related protein in human inflamed dental pulp.
        Eur Rev Med Pharmacol Sci. 2010; 14: 471-475
        • Jiang H.W.
        • Ling J.Q.
        • Gong Q.M.
        The expression of stromal cell-derived factor 1 (SDF-1) in inflamed human dental pulp.
        J Endod. 2008; 34: 1351-1354
        • Silva A.C.
        • Faria M.R.
        • Fontes A.
        • Campos M.S.
        • Cavalcanti B.N.
        Interleukin-1 beta and interleukin-8 in healthy and inflamed dental pulps.
        J Appl Oral Sci. 2009; 17: 527-532
        • Tecles O.
        • Laurent P.
        • Zygouritsas S.
        • et al.
        Activation of human dental pulp progenitor/stem cells in response to odontoblast injury.
        Arch Oral Biol. 2005; 50: 103-108
        • Fitzgerald M.
        • Chiego Jr., D.J.
        • Heys D.R.
        Autoradiographic analysis of odontoblast replacement following pulp exposure in primate teeth.
        Arch Oral Biol. 1990; 35: 707-715
        • Miura M.
        • Gronthos S.
        • Zhao M.
        • et al.
        SHED: stem cells from human exfoliated deciduous teeth.
        Proc Natl Acad Sci U S A. 2003; 100: 5807-5812
        • Bernardi L.
        • Luisi S.B.
        • Fernandes R.
        • et al.
        The isolation of stem cells from human deciduous teeth pulp is related to the physiological process of resorption.
        J Endod. 2011; 37: 973-979
        • da Silva Meirelles L.
        • Chagastelles P.C.
        • Nardi N.B.
        Mesenchymal stem cells reside in virtually all post-natal organs and tissues.
        J Cell Sci. 2006; 119: 2204-2213
        • Mouiseddine M.
        • Francois S.
        • Semont A.
        • et al.
        Human mesenchymal stem cells home specifically to radiation-injured tissues in a non-obese diabetes/severe combined immunodeficiency mouse model.
        Br J Radiol. 2007; 80: S49-S55
        • Ma D.
        • Ma Z.
        • Zhang X.
        • et al.
        Effect of age and extrinsic microenvironment on the proliferation and osteogenic differentiation of rat dental pulp stem cells in vitro.
        J Endod. 2009; 35: 1546-1553
        • Fakhry A.
        • Ratisoontorn C.
        • Vedhachalam C.
        • et al.
        Effects of FGF-2/-9 in calvarial bone cell cultures: differentiation stage-dependent mitogenic effect, inverse regulation of BMP-2 and noggin, and enhancement of osteogenic potential.
        Bone. 2005; 36: 254-266
        • Shiba H.
        • Mouri Y.
        • Komatsuzawa H.
        • et al.
        Enhancement of alkaline phosphatase synthesis in pulp cells co-cultured with epithelial cells derived from lower rabbit incisors.
        Cell Biol Int. 2003; 27: 815-823
        • Yan Q.
        • Sage E.H.
        SPARC, a matricellular glycoprotein with important biological functions.
        J Histochem Cytochem. 1999; 47: 1495-1506
        • Spoto G.
        • Fioroni M.
        • Rubini C.
        • Tripodi D.
        • Di Stilio M.
        • Piattelli A.
        Alkaline phosphatase activity in normal and inflamed dental pulps.
        J Endod. 2001; 27: 180-182
        • Horst O.V.
        • Horst J.A.
        • Samudrala R.
        • Dale B.A.
        Caries induced cytokine network in the odontoblast layer of human teeth.
        BMC Immunol. 2011; 12: 9
        • McLachlan J.L.
        • Smith A.J.
        • Bujalska I.J.
        • Cooper P.R.
        Gene expression profiling of pulpal tissue reveals the molecular complexity of dental caries.
        Biochim Biophys Acta. 2005; 1741: 271-281
        • Shi S.
        • Gronthos S.
        Perivascular niche of postnatal mesenchymal stem cells in human bone marrow and dental pulp.
        J Bone Miner Res. 2003; 18: 696-704
        • Gopinath S.D.
        • Rando T.A.
        Stem cell review series: aging of the skeletal muscle stem cell niche.
        Aging Cell. 2008; 7: 590-598
        • Schofield R.
        The relationship between the spleen colony-forming cell and the haemopoietic stem cell.
        Blood Cells. 1978; 4: 7-25
        • Huang A.H.
        • Snyder B.R.
        • Cheng P.H.
        • Chan A.W.
        Putative dental pulp-derived stem/stromal cells promote proliferation and differentiation of endogenous neural cells in the hippocampus of mice.
        Stem Cells. 2008; 26: 2654-2663
        • Gandia C.
        • Arminan A.
        • Garcia-Verdugo J.M.
        • et al.
        Human dental pulp stem cells improve left ventricular function, induce angiogenesis, and reduce infarct size in rats with acute myocardial infarction.
        Stem Cells. 2008; 26: 638-645
        • Yamauchi N.
        • Yamauchi S.
        • Nagaoka H.
        • et al.
        Tissue engineering strategies for immature teeth with apical periodontitis.
        J Endod. 2011; 37: 390-397
        • Spradling A.
        • Drummond-Barbosa D.
        • Kai T.
        Stem cells find their niche.
        Nature. 2001; 414: 98-104
        • Watt F.M.
        • Hogan B.L.
        Out of Eden: stem cells and their niches.
        Science. 2000; 287: 1427-1430
        • Alongi D.J.
        • Yamaza T.
        • Song Y.
        • et al.
        Stem/progenitor cells from inflamed human dental pulp retain tissue regeneration potential.
        Regen Med. 2010; 5: 617-631
        • Wang Z.
        • Pan J.
        • Wright J.T.
        • et al.
        Putative stem cells in human dental pulp with irreversible pulpitis: an exploratory study.
        J Endod. 2010; 36: 820-825
        • Yang X.
        • Zhang S.
        • Pang X.
        • Fan M.
        Proinflammatory cytokines induce odontogenic differentiation of dental pulp derived stem cells.
        J Cell Biochem. 2011; 113: 669-677
        • Goldberg M.
        • Farges J.C.
        • Lacerda-Pinheiro S.
        • et al.
        Inflammatory and immunological aspects of dental pulp repair.
        Pharmacol Res. 2008; 58: 137-147
        • Paula-Silva F.W.
        • Ghosh A.
        • Silva L.A.
        • Kapila Y.L.
        TNF-alpha promotes an odontoblastic phenotype in dental pulp cells.
        J Dent Res. 2009; 88: 339-344