Shanghai Journal of Stomatology ›› 2020, Vol. 29 ›› Issue (3): 242-249.doi: 10.19439/j.sjos.2020.03.004

• Original Articles • Previous Articles     Next Articles

Bone morphogenetic protein-2, -4, -6, -7 and -9 differentially mediated osteogenic differentiation of immortalized odontoblasts

LI Jing1,2,3, ZHANG Yi1,2,3, WANG Jin-hua1,2,3   

  1. 1. Stomatological Hospital of Chongqing Medical University. Chongqing 400015;
    2. Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences. Chongqing 401120;
    3. Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education. Chongqing 401120, China
  • Received:2020-02-20 Revised:2020-03-24 Online:2020-06-25 Published:2020-07-29

Abstract: PURPOSE: This study was aimed to investigate the mesenchymal stem cell (MSC) phenotypes of immortalized odontoblasts(iODs) and bone morphogenetic protein -2, -4, -6, -7, and -9 (BMPs) differentially regulate the mineralization of iODs. METHODS: ODs were immortalized by SV40 T antigen to establish iOD lineages, and the endogenous expression of BMPs was successively examined. Recombinant adenoviruses expressing BMPs and GFP were generated using Ad-Easy technology. The proliferation capability of iODs was examined using an MTT kit. MSC markers of iODs were examined by immunofluorescence. In vitro, semiquantitative RT-PCR, alkaline phosphatase(ALP) activity assay, matrix mineralization assay and oil red O staining assay were used to examine the osteogenic and adipogenic differentiation capabilities of iODs. Statistical significance among groups was analyzed by one-way analysis of variance and Scheffe's multiple comparison test was SPSS 21.0 software package. Finally, the volume and density of ectopic mineralized tissues formed in vivo were assessed by micro-CT and histological analysis. RESULTS: ODs can be efficiently immortalized by SV40 T antigen, and the resulting iODs maintained an excellent proliferative activity, expressed certain MSC markers and possessed multiple differentiation capabilities. BMP-2 and BMP-9 regulated iODs osteogenic differentiation better than BMP-4, -6, and -7. CONCLUSIONS: Our findings suggest that ODs and osteogenic growth factors such as BMP-2 and BMP-9 can be used as an efficacious strategy for bone tissue engineering.

Key words: Bone morphogenetic protein, Odontoblasts, Immortalized odontoblasts, Osteogenic differentiation

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