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

  • Original Article
  • Tissue Engineering
  • Published:
Tissue Engineering and Regenerative Medicine Aims and scope

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.

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Authors and Affiliations

  1. College of Medicine, Dongguk University, Goyang, Korea

    Jong Min Lee

  2. Department of Orthopaedics, Dongguk University Ilsan Hospital, Goyang, Korea

    Eun Ah Kim & Gun-Il Im

Authors
  1. Jong Min Lee
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  2. Eun Ah Kim
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  3. Gun-Il Im
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Correspondence to Gun-Il Im.

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Lee, J.M., Kim, E.A. & Im, GI. Healing of tibial and calvarial bone defect using Runx-2-transfected adipose stem cells. Tissue Eng Regen Med 12, 107–112 (2015). https://doi.org/10.1007/s13770-014-0070-3

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  • DOI: https://doi.org/10.1007/s13770-014-0070-3

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