Bone Morphogenetic Protein-2 Inhibits Differentiation and Mineralization of Cementoblasts in vitro
M. Zhao1,
J.E. Berry1, and
M.J. Somerman1,2,*
1 Department of Periodontics/Prevention/Geriatrics, University of Michigan School of Dentistry, 1011 N. University Avenue, Ann Arbor, MI 48109-1078, USA;
2 Department of Periodontics, School of Dentistry, University of Washington, D-322 Health Sciences Center, Box 356365, Seattle, WA 98195-6365, USA;
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Figure 1. Effect of BMP-2 on gene expression of cementoblasts. Cells were treated for 5 days with BMP-2 at concentrations up to 300 ng/mL, with or without AA, and Northern blot analyses were used to evaluate mineral-related gene markers. Results were normalized with 18S rRNA and are shown by bar graphs. Autoradiographs are representative of results obtained in three independent experiments.
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Figure 2. Effect of noggin on BMP-2-mediated expression of BSP and OCN. Cells were cultured in DMEM with 2% FBS, ± BMP-2 (300 ng/mL), ± AA (50 µg/mL), ± noggin (1 µg/mL) for 5 days, and Northern blot analysis was used to determine gene expression of BSP and OCN. Results were normalized against 18S rRNA and are shown by bar graphs. Autoradiographs are representative of results obtained in three independent experiments.
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Figure 3. Effect of BMP-2 on secretion of BSP and OCN protein. Cells at confluence were exposed to BMP-2 (0–300 ng/mL) with AA in serum-free media for 48 hrs. Conditioned media were collected and examined by Western blot assay with antibodies against BSP or OCN. This Fig. is representative of results obtained in three independent experiments.
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Figure 4. Effect of BMP-2 on cell-mediated mineral nodule formation. Cementoblasts were cultured in mineralizing media supplemented with BMP-2 (0–300 ng/mL) with or without noggin (1 µg/mL). To evaluate mineral nodule formation, we performed von Kossa staining on day 9.
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