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Effects of Cyclic Tensile Forces on the Expression of Vascular Endothelial Growth Factor (VEGF) and Macrophage-colony-stimulating Factor (M-CSF) in Murine Osteoblastic MC3T3-E1 Cells

Department of Orthodontics and Craniofacial Developmental Biology, Hiroshima University Graduate School of Biomedical Sciences, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, Japan;

View larger version (19K): [in a new window]   Figure 1. Changes in the expression of VEGF and M-CSF mRNAs in osteoblastic MC3T3-E1 cells stimulated by various elongations for 12 hrs. Means and standard deviations of the expression of VEGF and M-CSF mRNAs under several elongations are shown. The dotted line indicates the expression of VEGF and M-CSF mRNAs in the control group. It was shown that VEGF and M-CSF mRNAs in the experimental groups reached the maximum of 3.9- and 2.2-fold at 12% elongation. Each condition was prepared in triplicate, and experiments were performed 3 times. (*p < 0.05 vs. control group).

View larger version (28K): [in a new window]   Figure 2. Loading time changes in the expression of VEGF mRNA and protein concentration in osteoblastic MC3T3-E1 cells stimulated by 12% elongation. Means and standard deviations of the expression of VEGF mRNA and protein concentration under 12% elongation are shown. The dotted line indicates the expression of VEGF mRNA in the control group. The amount of VEGF mRNA in the experimental group significantly increased from 3 to 24 hrs after the loading compared with the control group. The protein concentration of VEGF in the experimental group increased time-dependently, showing significant differences compared with the control group at 3, 6, 12, and 24 hrs after the loading. Each condition was prepared in triplicate, and experiments were performed 3 times. (*p < 0.05 vs. control group).

View larger version (20K): [in a new window]   Figure 3. Loading time changes in the expression of M-CSF mRNA and protein concentration in osteoblastic MC3T3-E1 cells stimulated by 12% elongation. Means and standard deviations of the expression of M-CSF mRNA and protein concentration under 12% elongation are shown. The dotted line indicates the expression of M-CSF mRNA in the control group. The amount of M-CSF mRNA in the experimental group significantly increased after 12 and 24 hrs from the loading against the control group. The protein concentration of M-CSF in the experimental group increased time-dependently, showing significant differences compared with the control group at 6, 12, and 24 hrs after the loading. Each condition was prepared in triplicate, and experiments were performed 5 times. (*p < 0.05 vs. control group).

View larger version (17K): [in a new window]   Figure 4. Effects of Gd3+ and nifedipine on the expression of VEGF and M-CSF in osteoblastic MC3T3-E1 cells stimulated by 12% elongation for 12 hrs (A). The protein concentrations of VEGF (B) and M-CSF (C) in the conditioned medium from MC3T3-E1 cells subjected to 12% elongation at 30 cycles/min for 12 hrs with or without Gd3+. (A) Means and standard deviations of the expression of VEGF and M-CSF mRNA are shown. The dotted line indicates the expression of VEGF and M-CSF mRNAs in the control group. The expression of VEGF mRNA was significantly reduced in the 100-µm Gd3+ treatment group but not in the 10-µm nifedipine group. The 10- and 100-µm Gd3+ treatment groups showed a significant decrease in the M-CSF mRNA, but there was no corresponding decrease in the difedipine group. Treatment with 100 µm Gd3+ showed no effect on control cells in the expressions of both VEGF and M-CSF mRNAs. (B,C) Means and standard deviations of the protein concentrations of VEGF and M-CSF were measured by a quantitative sandwich enzyme immunoassay technique. Significant decreases in VEGF and M-CSF concentrations were found in the 10- and 100-µm Gd3+ treatment groups. Each condition was prepared in triplicate, and experiments were performed 5 times. (*p < 0.05).