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Enamel Matrix Derivative Gel Stimulates Signal Transduction of BMP and TGF-ß

¹ Department of Periodontics and Endodontics, and
² Department of Biochemistry, School of Dental Medicine, Tsurumi University, 2-1-3 Tsurumi, Tsurumi-ku, Yokohama 230-8501, Japan;
³ Division of Pathophysiology, Research Center for Genomic Medicine, Saitama Medical School, 1397-1 Yamane Hidaka, Saitama 350-1241, Japan; and
⁴ Department of Biologic and Materials Science, School of Dentistry, University of Michigan, 1210 Eisenhower Place, Ann Arbor, MI 48108, USA;

View larger version (30K): [in a new window]   Figure 1. Biological activity induced by unfractionated Emd-Gel. Mv1Lu cells were transfected with PAI-1 gene, then treated with (A) TGF-ß1 (0.05, 0.1, 0.2, 0.4, 0.6 ng/mL) and (B) Emd-Gel (10, 20, 30, 40, 50 µg/mL). C2C12 cells were transfected with Id1 gene, and then treated with (C) BMP2 (50, 100, 150, 200, 250 ng/mL) and (D) Emd-Gel (25, 50, 75, 100, 125 µg/mL). Luciferase activity was determined after an overnight incubation. Data are means ± SE of 4 culture wells. *p < 0.05, **p < 0.001.

View larger version (47K): [in a new window]   Figure 2. Fractionation of Emd-Gel from Sephadex G-100 column. (A) The chromatogram represents absorbance at 280 nm. (B) SDS-PAGE profiles of the separated fractions stained with CBB. Molecular weights (Bio-Rad Low Range Standards, Hercules, CA, USA) are shown in left margin.

View larger version (33K): [in a new window]   Figure 3. TGF-ß and BMP activities induced by fractionated Emd-Gel. Mv1Lu cells were transfected with PAI-1 gene, and C2C12 cells were transfected with Id1 gene. These cells were then treated with fractionated Emd-Gel. Luciferase activity was determined after an overnight incubation. Data are means ± SE of 4 culture wells. *p < 0.05.

View larger version (19K): [in a new window]   Figure 4. Effect of TGF-ß on signal transduction of BMP. C2C12 cells were transfected with Id1 gene, then treated with (A) 100 ng/mL BMP2 and 10 ng/mL TGF-ß1, and with (B) BMP-like fraction (fractions 4-6) in Emd-Gel added to TGF-ß and TGF-ß-like fraction (fractions 8-13) in Emd-Gel. Luciferase activity was measured after an overnight incubation. Data are means ± SE of 4 culture wells. *p < 0.05, **p < 0.001.