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Noggin Blocks Osteoinductive Activity of Porcine Enamel Extracts

¹ Section of Periodontology, Department of Hard Tissue Engineering, Graduate School, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8549, Japan; and
² Department of Biochemistry, School of Dental Medicine, Tsurumi University, 2-1-3 Tsurumi, Tsurumi-ku, Yokohama, Kanagawa 230-8501, Japan;

View larger version (59K): [in a new window]   Figure 1. Elution profile of porcine enamel extracts from Sephadex G-100 column and ALP activity of ST2 cells induced by each fraction. (A) The chromatogram represents absorbance at 280 nm. Bars show ALP activities of ST2 cells exposed to G-100 fractions. Data are means ± SE of three culture wells. SDS-PAGE profiles of fractions of I to V (B) and G-100 fractions from No. 36 to No. 50 (C) stained with CBB. Molecular weights (Bio-Rad Low Range Standards, Hercules, CA, USA) are shown in left margin.

View larger version (38K): [in a new window]   Figure 2. (A) Effect of 50 µg/mL OFE on osteocalcin, bone sialoprotein (BSP), and alkaline phosphatase (ALP) mRNA expression in cultures of ST2 cells. Total RNA of ST2 cells incubated with or without OFE was collected on days 1, 4, 7, 11, and 14. Each culture was incubated with 50 µM ascorbic acid, 10 mM ß-glycerophosphate, and 200 nM retinoic acid. (B) Nodule formation induced by OFE. OFE was added to medium at concentrations of 0, 12.5, 25, and 50 µg/mL. Nodule cultures were stained with 2% alizarin red S on day 15. The stained area was quantified by NIH Imaging. Data are means ± SE of three culture wells. Significantly different from culture without OFE at *p < 0.05, **p < 0.001.

View larger version (17K): [in a new window]   Figure 3. The influence of noggin incubated with OFE on ALP activity of ST2 cells. OFE (40 µg/mL) and noggin (1, 10, 100, or 300 ng/mL) were incubated for 1 hr and then added to the cells. ALP activity was measured 72 hrs later. Data are means ± SE of three culture wells. Significantly different from culture without noggin at *p < 0.05.

View larger version (57K): [in a new window]   Figure 4. Dot-blot analysis of BMP2/4 protein in OFE. Lane 1, 50 µg of OFE; lane 2, 500 ng of rhBMP-2 (Genzyme-Techne, USA; positive control); and lane 3, 50 µg of BSA (Sigma, St. Louis, MO, USA; negative control). Each sample was diluted as shown in left margin.