Biodentine induces immortalized murine pulp cell differentiation into odontoblast-like cells and stimulates biomineralization.

INTRODUCTION Biodentine (Septodont, Saint Maur des Faussés, France), a new tricalcium silicate-based cement, has recently been commercialized and advertised as a bioactive material. Its clinical application and physical properties have been widely described, but, so far, its bioactivity and biological effect on pulp cells have not been clearly shown. Thus, the aim of this study was to evaluate the biological effect of Biodentine on immortalized murine pulp cells (OD-21). METHODS OD-21 cells were cultured with or without Biodentine. Cell proliferation was determined by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) colorimetric assay after 2, 3, and 5 days of stimulation. The expression of several biomolecular markers was analyzed to screen differentiation pathways, both on a gene level with Real-time reverse transcription polymerase chain reaction and on a protein level by measuring alkaline phosphatase activity. Alizarin red staining was used to assess and quantify biomineralization. RESULTS The expression patterns of several genes confirmed the differentiation of OD-21 cells into odontoblasts during the period of cell culture. Our results suggest that Biodentine is bioactive because it increased OD-21 cell proliferation and biomineralization in comparison with controls. CONCLUSIONS Because of its bioactivity, Biodentine can be considered as a suitable material for clinical indications of dentin-pulp complex regeneration, such as direct pulp capping.

[1]  C. Rahiotis,et al.  Parallels between Tooth Development and Repair: Conserved Molecular Mechanisms following Carious and Dental Injury , 2004, Journal of dental research.

[2]  Howard C. Tenenbaum,et al.  Dexamethasone induces proliferation and terminal differentiation of osteogenic cells in tissue culture , 1986, The Anatomical record.

[3]  Hai Zhang,et al.  Direct contact with mineral trioxide aggregate activates and differentiates human dental pulp cells. , 2011, Journal of endodontics.

[4]  H. Lesot,et al.  Reactionary dentinogenesis. , 1995, The International journal of developmental biology.

[5]  Y. H. Wang,et al.  Runx2, Osx, and Dspp in Tooth Development , 2009, Journal of dental research.

[6]  Jiang Chang,et al.  The self-setting properties and in vitro bioactivity of tricalcium silicate. , 2005, Biomaterials.

[7]  Yuan Yuan,et al.  In vitro osteoblast-like and endothelial cells' response to calcium silicate/calcium phosphate cement , 2010, Biomedical materials.

[8]  K. Langeland,et al.  Capping of monkey pulps with Dycal and a Ca-eugenol cement. , 1981, Oral surgery, oral medicine, and oral pathology.

[9]  J. Ferracane,et al.  Can interaction of materials with the dentin-pulp complex contribute to dentin regeneration? , 2010, Odontology.

[10]  Hai Zhang,et al.  Effects of mineral trioxide aggregate on human dental pulp cells after pulp-capping procedures. , 2010, Journal of endodontics.

[11]  R. D'Souza,et al.  Molecular Insights into the Lineage-specific Determination of Odontoblasts: The Role of Cbfa1 , 2001, Advances in dental research.

[12]  A. Berdal,et al.  Evaluation of a new laboratory model for pulp healing: preliminary study. , 2008, International endodontic journal.

[13]  U. Stratmann,et al.  Direct pulp capping with mineral trioxide aggregate: an immunohistologic comparison with calcium hydroxide in rodents. , 2010, Journal of endodontics.

[14]  P. Planells,et al.  Dentin bridge formation after mineral trioxide aggregate (MTA) pulpotomies in primary teeth. , 2005, American journal of dentistry.

[15]  A. Berdal,et al.  Differential Expression and Activity of Tissue-nonspecific Alkaline Phosphatase (TNAP) in Rat Odontogenic Cells In Vivo , 1999, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[16]  T. Latifi,et al.  Reciprocal Temporospatial Patterns of Msx2 and Osteocalcin Gene Expression During Murine Odontogenesis , 1998, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[17]  Qiang Zhu,et al.  Effects of root-end filling materials on fibroblasts and macrophages in vitro. , 2003, Oral surgery, oral medicine, oral pathology, oral radiology, and endodontics.

[18]  A. Berdal,et al.  Molecular characterization of young and mature odontoblasts. , 2009, Bone.

[19]  G. Karsenty,et al.  Osf2/Cbfa1: A Transcriptional Activator of Osteoblast Differentiation , 1997, Cell.

[20]  R. K. Sübay,et al.  Tunnel defects in dentin bridges: their formation following direct pulp capping. , 1996, Operative dentistry.

[21]  G. Stein,et al.  Dlx3 Transcriptional Regulation of Osteoblast Differentiation: Temporal Recruitment of Msx2, Dlx3, and Dlx5 Homeodomain Proteins to Chromatin of the Osteocalcin Gene , 2004, Molecular and Cellular Biology.

[22]  Toshihisa Komori,et al.  Regulation of osteoblast differentiation by transcription factors , 2006, Journal of cellular biochemistry.

[23]  L. Baume The biology of pulp and dentine. A historic, terminologic-taxonomic, histologic-biochemical, embryonic and clinical survey. , 1980, Monographs in oral science.

[24]  H. Lesot,et al.  Induction and regulation of crown dentinogenesis: embryonic events as a template for dental tissue repair? , 2001, Critical reviews in oral biology and medicine : an official publication of the American Association of Oral Biologists.

[25]  S. K. Zaidi,et al.  Runx2 control of organization, assembly and activity of the regulatory machinery for skeletal gene expression , 2004, Oncogene.

[26]  L. Baume I. Stepping-Stones to Pulpodentinal Biology , 1980 .

[27]  D. Prockop,et al.  An Alizarin red-based assay of mineralization by adherent cells in culture: comparison with cetylpyridinium chloride extraction. , 2004, Analytical biochemistry.

[28]  Chia-Che Ho,et al.  In vitro bioactivity and biocompatibility of dicalcium silicate cements for endodontic use. , 2009, Journal of endodontics.

[29]  U. Schröder Effects of Calcium Hydroxide-containing Pulp-capping Agents on Pulp Cell Migration, Proliferation, and Differentiation , 1985, Journal of dental research.

[30]  M. Leite,et al.  The effect of ionic products from bioactive glass dissolution on osteoblast proliferation and collagen production. , 2004, Biomaterials.

[31]  M. Larmas Pre-odontoblasts, Odontoblasts, or “Odontocytes” , 2008, Journal of dental research.

[32]  F. Mcdonald,et al.  Runx2 and dental development. , 2006, European journal of oral sciences.

[33]  M. Torabinejad,et al.  Using mineral trioxide aggregate as a pulp-capping material. , 1996, Journal of the American Dental Association.

[34]  G. Belibasakis,et al.  The dentinogenic effect of mineral trioxide aggregate (MTA) in short-term capping experiments. , 2002, International endodontic journal.

[35]  J. Wataha,et al.  Cloned 3T6 cell line from CD-1 mouse fetal molar dental papillae. , 1998, Connective tissue research.

[36]  D. Pashley Dynamics of the pulpo-dentin complex. , 1996, Critical reviews in oral biology and medicine : an official publication of the American Association of Oral Biologists.