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KMID : 0361620070420030386
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Journal of the Korean Orthopaedic Association
2007 Volume.42 No. 3 p.386 ~ p.394
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Osteoinduction using by Human BMP-2-physio-functional Bioactive Ceramics
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Kim Hyang
Moon Seong-Hwan
Lee Hwan-Mo
Lee Jeong-Wook
Kim Hyun-Min
Park Min
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Abstract
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Purpose : To assess the properties and the osteogenic potency of the calcium phosphate-recombinant human morphogenetic protein-2 (CaP-rhBMP-2 composite) on glass-ceramics.
Materials and Methods : Bioactive glass-ceramics,as a scaffold, and a calcium phosphate (CaP) solution (pH7.4) were prepared. Recombinant human bone morphogenetic protein-2 (rhBMP-2) was purified from CHO-K1 cells by transfecting the cells with BMP-2 cDNA. The glass-ceramics were soaked for 3 days at room temperature in saline, a CaP only solution, and a CaP solution containing rhBMP-2. Scanning electron microscopy (SEM), Fourier transform infrared reflection spectroscopy (FT-IR), thin film X-ray diffraction (TF-XRD) and immunofluorescent staining (IF) of the anti-human BMP-2 to composite-coated scaffold were performed to verify the characterization of the scaffold surface. In addition, RT-PCR of osteogenic marker gene and SEM photography were performed after adhering the mouse preosteoblast MC3T3-E1 cells in order to assess the osteoinductivity.
Results : CaP-rhBMP-2 composite was coated on the surface of glass-ceramics, as confirmed by SEM, FT-IR, TF-XRD spectrum, and IF. The CaP-rhBMP-2 composite on the glass-ceramic showed a globular shape covered with fine spikes while the CaP on the glass-ceramic showed a fine spike structure on the flat glass surface. The expression of collagen type I and alkaline phosphatase mRNAs had increased 4 hours after cell seeding. In addition, the level of osteocalcin mRNA expression had increased significantly by 3 days in the CaP-rhBMP-2 composite compared with the control and CaP group. The SEM photographs showed more active filopodia formation in the CaP-rhBMP-2 composite than the other groups. There was extensive newly synthesized extracellular matrix around the osteoblasts and CaP-rhBMP-2 composite nodule.
Conclusion : The application of CaP-rhBMP-2 composite-surface coating technique on bioactive glass-ceramic is a powerful tool for osteoinduction.
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KEYWORD
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Calcium phosphate-rhBMP-2 composite, Surface coating technique, Osteoinductivity
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