title: Wnt3a enhances bone morphogenetic protein 9-induced osteogenic differentiation of C3H10T1/2 cells.
abstract: BACKGROUND
  Bone morphogenetic protein 9 (BMP9) and Wnt/β-catenin signaling pathways are able to induce osteogenic differentiation of mesenchymal stem cells (MSCs), but the role of Wnt/β-catenin signaling pathway in BMP9-induced osteogenic differentiation is not well understood. Thus, our experiment was undertaken to investigate the interaction between BMP9 and Wnt/β-catenin pathway in inducing osteogenic differentiation of MSCs.
  
  
  METHODS
  C3H10T1/2 cells were infected with recombinant adenovirus expressing BMP9, Wnt3a, and BMP9+Wnt3a. ALP, the early osteogenic marker, was detected by quantitative and staining assay. Later osteogenic marker, mineral calcium deposition, was determined by Alizarin Red S staining. The expression of osteopotin (OPN), osteocalcin (OC), and Runx2 was analyzed by Real time PCR and Western blotting. In vivo animal experiment was carried out to further confirm the role of Wnt3a in ectopic bone formation induced by BMP9.
  
  
  RESULTS
  The results showed that Wnt3a enhanced the ALP activity induced by BMP9 and increased the expressions of OC and OPN, with increase of mineral calcium deposition in vitro and ectopic bone formation in vivo. Furthermore, we also found that Wnt3a increased the level of Runx2, an important nuclear transcription factor of BMP9.
  
  
  CONCLUSIONS
  Canonical Wnt/β-catenin signal pathway may play an important role in BMP9-induced osteogenic differentiation of MSCs, and Runx2 may be a linkage between the two signal pathways.
authors.affiliation: Department of Orthopedics, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Orthopedics, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
authors.forename: Liang-Bo
authors.initials: LB
authors.lastname: Lin
authors.affiliation: Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Dao-Jing
authors.initials: DJ
authors.lastname: Xu
authors.affiliation: Department of Orthopedics, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
authors.forename: Rong-Fu
authors.initials: RF
authors.lastname: Chen
authors.affiliation: Department of Orthopedics, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
authors.forename: Ji-Xiang
authors.initials: JX
authors.lastname: Tan
authors.affiliation: Department of Orthopedics, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
authors.forename: Xi
authors.initials: X
authors.lastname: Liang
authors.affiliation: Department of Orthopedics, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
authors.forename: Ning
authors.initials: N
authors.lastname: Hu
authors.affiliation: Department of Orthopedics, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China. Email: huangwei68@263.net.
authors.forename: Wei
authors.initials: W
authors.lastname: Huang
mesh_terms: D015153:Blotting, Western; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology*; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics / Q000378:metabolism; D055427:Growth Differentiation Factor 2 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D015675:Osteocalcin / Q000235:genetics / Q000378:metabolism; D010012:Osteogenesis / Q000235:genetics / Q000502:physiology*; D060509:Wnt3A Protein / Q000235:genetics / Q000378:metabolism*
chemical_list: D050659:Core Binding Factor Alpha 1 Subunit; D055427:Growth Differentiation Factor 2; D060509:Wnt3A Protein; D015675:Osteocalcin