IADR Abstract Archives

Promotion of rat calvarial defect healing by induced bone-like tissue

Recent experiments have attempted to induce bone from immature muscular tissue(IMT) in vitro using crude BMP and e-PTFE as a scaffold. This process induces bone-like tissue; however, this induced bone-like tissue has ossification potential within 2 weeks after implantation. Meanwhile, recombinant human BMP(rhBMP)-2, -4 and -7 has osteogenic potential both in vitro and in vivo.

Objectives: The aim of this study was to induce bone-like tissue from IMT in vitro by rhBMP-2, rhBMP-4 and rhBMP-7 first, and then implant this tissue into a calvarial defect in rats and finally assess the healing.

Methods : Commercially available rhBMP-2, rhBMP-4 and rhBMP-7 were used in this experiment. IMTs were extracted from the 20-day Sprague-Dawley embryonic rats and placed on e-PTFE with 10ng/μl each of rhBMP-2, -4, and -7, and cultured for 2 weeks. The cultured tissues were analyzed by μCT, histological observation and EPMA. RT-PCR was performed to examine the gene expression of Runx2, collagen type I, osteopontin and osteocalcin on days 7 and 14. Furthermore, the specimens were implanted into calvarial defects in Sprague-Dawley rats for up to 3 weeks, and then evaluated by μCT and histological observation.

Results: Relatively strong radiopacity was observed by μCT at 2 weeks after culture. Mineral deposition was observed by Von Kossa staining. The expression of all osteoblastic marker genes was confirmed on both days 7 and 14. Ca and P were detected in the extracellular matrix by EPMA. In addition, the strong radiopacity and partial ossification were confirmed at 1 week after implantation, and dominant new bone was observed after 2 weeks in the defect area.

Conclusion: The rhBMPs differentiated IMT into bone-like tissue in vitro, and this induced bone-like tissue has ossification potential and promotes healing of calvarial defects. These results suggest that IMT will be an effective tissue source for bone tissue engineering.

IADR/LAR General Session
2012 IADR/LAR General Session (Iguaçu Falls, Brazil)
Iguaçu Falls, Brazil
2012
69
Prosthodontics Research
  • Hayashi, Tatsuhide  ( Aichi Gakuin University, Nagoya, N/A, Japan )
  • Kobayashi, Shuichiro  ( Aichi Gakuin University, Nagoya, N/A, Japan )
  • Asai, Takafumi  ( Aichi Gakuin University, Nagoya, N/A, Japan )
  • Mieki, Akimichi  ( Aichi Gakuin University, Nagoya, N/A, Japan )
  • Kataoka, Hiroyasu  ( Aichi Gakuin University, Nagoya, N/A, Japan )
  • Uematsu, Yasuaki  ( Aichi Gakuin University, Nagoya, N/A, Japan )
  • Kawai, Tatsushi  ( Aichi Gakuin University, Nagoya, N/A, Japan )
    • Oral Session
    Frechette Competition
    06/20/2012