Stimulation of reparative dentin formation by ex vivo gene therapy using dental pulp stem cells electrotransfected with growth/differentiation factor 11 (Gdf11)

Misako Nakashima, Koichiro Iohara, Masaki Ishikawa, Masataka Ito, Atsushi Tomokiyo, Takamasa Tanaka, Akifumi Akamine

研究成果: Contribution to journalArticle査読

66 被引用数 (Scopus)

抄録

Dental pulp progenitor/stem cells have the capacity to differentiate into odontoblasts and they provide a potential for dentin repair and regeneration by gene therapy. To develop a successful ex vivo gene therapy to induce reparative dentin formation rapidly and effectively after treatment of caries, we developed a three-dimensional pellet culture system of pulp cells electrotransfected with growth/differentiation factor 11 (Gdf11). The viability after electrotransfection was more than 85%, and the efficiency was about 70% as determined by flow cytometry. After 10 days of culture, the total amount of type I and type III collagen was 3-fold higher in the pEGFP-Gdf11-transfected pellet than in the control. Real-time RT-PCR analysis demonstrated that the expression of markers of odontoblast differentiation (alkaline phosphatase, dentin matrix protein 1 [Dmp1], dentin sialophosphoprotein [Dspp], enamelysin, and phosphate-regulating gene with homologies to endopeptidases on X-chromosome [Phex]) was increased in the pEGFP-Gdf11-transfected pellet compared with the control on day 14. On the basis of this in vitro evaluation, an in vivo investigation in the dog was performed. Autogenous transplantation of Gdf11-transfected cells cultured as a pellet on amputated pulp stimulated reparative dentin formation. Thus, Gdf11 gene therapy may be potentially used in endodontic treatment in dentistry.

本文言語英語
ページ(範囲)1045-1053
ページ数9
ジャーナルHuman Gene Therapy
15
11
DOI
出版ステータス出版済み - 11 1 2004

All Science Journal Classification (ASJC) codes

  • 分子医療
  • 分子生物学
  • 遺伝学

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