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In vitro Effects of Enamel Matrix Proteins on Rat Bone Marrow Cells and Gingival Fibroblasts

¹ Departments of Oral Biology and
² Periodontology, The Maurice and Gabriela Goldschleger School of Dental Medicine, Tel Aviv University, Tel Aviv 69978, Israel;

View larger version (46K): [in a new window]   Figure 1. Nodule formation by BMSC. (A) Representative wells stained with the Von-Kossa method showing more mineralized nodules in cultures treated with Emdogain (EMD) at 25 µg/mL. (B) Quantitation of the number of nodules/well (mean ± SE, N = 6/group). Only EMD at 25 µg/mL increased nodule number. ***P < 0.005 (EMD at 25 µg/mL vs. control or 100 µg/mL). (C) Quantitation of the dish area covered with mineralized nodules (mean ± SE, N = 6/group). Only EMD at 25 µg/mL increased nodule area. ***P < 0.005 (EMD at 25 µg/mL vs. control or 100 µg/mL).

View larger version (20K): [in a new window]   Figure 2. (A) Alkaline phosphatase (ALP) activity of BMSC is enhanced by Emdogain (EMD) at 25 µg/mL (mean ± SE, N = 6/group). **P < 0.01 (EMD vs. control). (B) Quantitation of the number of nodules/well (mean ± SE, N = 6/group). EMD (25 µg/mL) was added immediately upon seeding (Time 0) or 48 hrs later, after removal of non-adherent cells. Only EMD at time 0 increased nodule formation. ***P < 0.005 (EMD at Time 0 vs. control or EMD at 48 hrs). (C) EMD (25 µg/mL) increases the number of adherent BMSC at 48 hrs (mean ± SE, N = 6/group). ***P < 0.005 (EMD vs. control). (D) EMD (25 µg/mL) increases the protein content of BMSC cultures on day 10 (mean ± SE, N = 6/group). *P < 0.05 (EMD vs. control).

View larger version (40K): [in a new window]   Figure 3. GF cultures. (A) Representative wells after Von-Kossa staining showing no mineralization but darker color of the matrix, with a greater effect of Emdogain (EMD) at 100 µg/mL than at 25 µg/mL. (B) Average optical density in arbitrary units of the GF matrix. Since darker color is associated with lower OD values (255 = white, 0 = black), the matrix of GF cells treated with EMD stains more intensely (mean ± SE, N = 6/group). ***P < 0.005 vs. control. (C) The protein content of GF cultures increases with the EMD concentration (mean ± SE, N = 6/group). *P < 0.05, ***P < 0.005 (vs. control).

View larger version (12K): [in a new window]   Figure 4. Emdogain (EMD, 100 µg/mL) increases GF cell number, determined on days 2, 5, or 12 (mean ± SE, N = 6/group). *P < 0.05; **P < 0.01; ***P < 0.005 (vs. the respective control).