title: In vitro evaluation of the odontogenic potential of mouse undifferentiated pulp cells.
abstract: The aim of this study was to evaluate the odontogenic potential of undifferentiated pulp cells (OD-21 cell line) through chemical stimuli in vitro. Cells were divided into uninduced cells (OD-21), induced cells (OD-21 cultured in supplemented medium/OD-21+OM) and odontoblast-like cells (MDPC-23 cell line). After 3, 7, 10 and 14 days of culture, it was evaluated: proliferation and cell viability, alkaline phosphatase activity, total protein content, mineralization, immunolocalization of dentin matrix acidic phosphoprotein 1 (DMP1), alkaline phosphatase (ALP) and osteopontin (OPN) and quantification of genes ALP, OSTERIX (Osx), DMP1 and runt-related transcription factor 2 (RUNX2) through real-time polymerase chain reaction (PCR). Data were analyzed by Kruskal-Wallis and Mann-Whitney U tests (p<0.05). There was a decrease in cell proliferation in OD-21 + OM, whereas cell viability was similar in all groups, except at 7 days. The amount of total protein was higher in group OD-21 + OM in all periods; the same occurred with ALP activity after 10 days when compared with OD-21, with no significant differences from the MDPC-23 group. Mineralization was higher in OD-21+OM when compared with the negative control. Immunolocalization demonstrated that DMP1 and ALP were highly expressed in MDPC-23 cells and OD-21 + OM cells, whereas OPN was high in all groups. Real-time PCR revealed that DMP1 and ALP expression was higher in MDPC-23 cell cultures, whereas RUNX2 was lower for these cells and higher for OD-21 negative control. Osx expression was lower for OD-21 + OM. These results suggest that OD-21 undifferentiated pulp cells have odontogenic potential and could be used in dental tissue engineering.
authors.affiliation: Ribeirão Preto Dental School, University of São Paulo, Ribeirão Preto, SP, Brazil.
authors.forename: Mayara Sgarbi
authors.initials: MS
authors.lastname: Semeghini
authors.forename: Roger Rodrigo
authors.initials: RR
authors.lastname: Fernandes
authors.forename: Daniela Thomazatti
authors.initials: DT
authors.lastname: Chimello
authors.forename: Fabíola Singaretti
authors.initials: FS
authors.lastname: de Oliveira
authors.forename: Karina Fittipaldi
authors.initials: KF
authors.lastname: Bombonato-Prado
mesh_terms: D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D002452:Cell Count; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D050659:Core Binding Factor Alpha 1 Subunit / Q000032:analysis; D003470:Culture Media; D003782:Dental Pulp / Q000166:cytology*; D016326:Extracellular Matrix Proteins / Q000032:analysis; D051379:Mice; D009804:Odontoblasts / Q000187:drug effects; D009805:Odontogenesis / Q000502:physiology*; D053495:Osteopontin / Q000032:analysis; D010750:Phosphoproteins / Q000032:analysis; D011506:Proteins / Q000032:analysis; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000076182:Sp7 Transcription Factor; D013997:Time Factors; D014074:Tooth Calcification / Q000187:drug effects; D014157:Transcription Factors / Q000032:analysis
chemical_list: D050659:Core Binding Factor Alpha 1 Subunit; D003470:Culture Media; C513309:DMP1 protein, human; D016326:Extracellular Matrix Proteins; D010750:Phosphoproteins; D011506:Proteins; C493709:RUNX2 protein, human; D000076182:Sp7 Transcription Factor; C478163:SP7 protein, human; D014157:Transcription Factors; D053495:Osteopontin; D000469:Alkaline Phosphatase