title: Effects of WNT10A on proliferation and differentiation of human dental pulp cells.
abstract: BACKGROUND
  Wingless-type MMTV integration site family, member 10A (WNT10A) plays crucial roles in odontogenesis. The aim of this study was to investigate the effects of WNT10A on human dental pulp cells (DPCs), which contain a mixed population of cells, including stem and progenitor cells, and participate in dentin repair or dentin-pulp regeneration.
  
  
  METHODS
  Healthy human premolars extracted for orthodontic reasons were used as a study model. The expression of WNT10A protein in dental pulp was determined by immunohistochemistry. The messenger RNA expression of WNT10A and Wnt-related genes was analyzed by semiquantitative reverse-transcription polymerase chain reaction. DPCs were enzymatically separated from pulp tissues, cultured, and passaged. The biological effects of WNT10A on DPCs were investigated using recombinant lentivirus encoding WNT10A complementary DNA. WNT10A-induced changes in DPC proliferation were assessed by methyltetrazolium assay and flow cytometry. In order to determine the effects of WNT10A on DPC differentiation, the activity of alkaline phosphatase (ALP), an early marker of odontoblastic differentiation, was assessed using an ALP activity assay kit, and the expression levels of odontoblast-specific genes, including DSPP, DMP1, ALP, and COL1A1, were detected by quantitative polymerase chain reaction and Western blot.
  
  
  RESULTS
  WNT10A protein was clearly identified in the cytoplasm of DPCs. Semiquantitative reverse-transcription polymerase chain reaction indicated the expression of WNT10A and Wnt-related genes in pulp tissues as well as in passaging DPCs. Lentiviral overexpression of WNT10A enhanced proliferation of DPCs and down-regulated ALP activity and the expression of odontoblast-specific genes.
  
  
  CONCLUSIONS
  WNT10A promotes the proliferation of DPCs and negatively regulates their odontoblastic differentiation.
authors.affiliation: Department of Cariology and Endodontology, School and Hospital of Stomatology, Peking University, Beijing, People's Republic of China.
authors.forename: Zhichun
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Department of Cariology and Endodontology, School and Hospital of Stomatology, Peking University, Beijing, People's Republic of China.
authors.forename: Qingru
authors.initials: Q
authors.lastname: Guo
authors.affiliation: Department of Cariology and Endodontology, School and Hospital of Stomatology, Peking University, Beijing, People's Republic of China.
authors.forename: Hua
authors.initials: H
authors.lastname: Tian
authors.affiliation: Department of Cariology and Endodontology, School and Hospital of Stomatology, Peking University, Beijing, People's Republic of China.
authors.forename: Ping
authors.initials: P
authors.lastname: Lv
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Peking University, Beijing, People's Republic of China.
authors.forename: Chunyan
authors.initials: C
authors.lastname: Zhou
authors.affiliation: Department of Cariology and Endodontology, School and Hospital of Stomatology, Peking University, Beijing, People's Republic of China. Electronic address: kqxjgao@bjmu.edu.cn.
authors.forename: Xuejun
authors.initials: X
authors.lastname: Gao
mesh_terms: D000293:Adolescent; D000328:Adult; D000469:Alkaline Phosphatase / Q000032:analysis; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D024042:Collagen Type I / Q000032:analysis; D000090602:Collagen Type I, alpha 1 Chain; D003593:Cytoplasm / Q000737:chemistry; D003782:Dental Pulp / Q000737:chemistry / Q000166:cytology* / Q000187:drug effects; D016326:Extracellular Matrix Proteins / Q000032:analysis; D006801:Humans; D009804:Odontoblasts / Q000187:drug effects; D010750:Phosphoproteins / Q000032:analysis; D012795:Sialoglycoproteins / Q000032:analysis; D013234:Stem Cells / Q000187:drug effects; D051153:Wnt Proteins / Q000032:analysis / Q000494:pharmacology*; D055815:Young Adult; D000099311:Dentin Sialophosphoprotein
chemical_list: D000469:Alkaline Phosphatase; D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D016326:Extracellular Matrix Proteins; D010750:Phosphoproteins; D012795:Sialoglycoproteins; D051153:Wnt Proteins; D000099311:Dentin Sialophosphoprotein; C513309:DMP1 protein, human; C484142:WNT10A protein, human
keywords: Dentin-pulp regeneration; WNT10A; dentin repair; human dental pulp cells