Healing of tibial and calvarial bone defect using Runx-2-transfected adipose stem cells

Jong Min Lee, Eun Ah Kim, Gun Il Im

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

The purpose of this study was to test the effect of Runx-2-transfected hASCs to heal the defect created on proximal tibiae and calvaria of immunosuppressed rats. Three kinds of hASCs (untransfected, pECFP-transfected ASCs or Runx-2-transfected ASCs) were cultured under osteogenic medium. Osteoblastic differentiation was measured by ALP staining on day 7 and osteoid matrix formation was observed by alizarin red staining on day 14 after osteogenic induction. Osteogenic potential in long bone defects were tested via 6 mm-sized circular defect on proximal tibiae of 9 immunosuppressed rats. Untransfected ASCs, pECFP-transfected ASCs or Runx-2-transfected ASCs embedded in fibrin scaffold were implanted in the defect (N=3 in each group). In order to assess the in vivo osteogenic capability of Runx-2-transfected ASC in intramembanous ossification, two critical size bone defects were created on parietal bone of 12 immunosuppressed rats. The defects were filled with fibrin scaffold containing pECFP-transfected ASCs, Runx-2-transfected ASCs or no cell (N=4 in each group). Runx-2 transfected ASCs showed much stronger activity of ALP and greater formation of osteoid matrix compared with untransfected ASCs or pECFP-transfected ASCs 7 and 14 after osteo-induction, respectively. When the volume of regenerated bone was compared from gross examination and radiographs after 5 weeks in the proximal tibial defect model, the defects treated with Runx-2-transfected ASCs had the greatest area of healed bone compared with other groups. In the calvarial defect model, Runx-2-transfected ASCs had significantly increased area healed with bone (p<0.05) as well as better quality of regenerated bone compared with defects which was treated with untransfected ASCs from gross and micro-CT examination 8 weeks after implantation. The implanted human cells persisted in the newly regenerated bone in defects treated with pECFP-ASCs and Runx-2-transfected ASCs. In conclusion, Runx-2-transfection significantly increased the osteogenic potential of ASCs in the in vivo orthotopic models.

Original languageEnglish
Pages (from-to)107-112
Number of pages6
JournalTissue Engineering and Regenerative Medicine
Volume12
Issue number2
DOIs
StatePublished - 1 Apr 2015

Keywords

  • adipose stem cell
  • bone defect
  • non-viral transfection
  • Runx-2

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