Effect Of Static Magnetic Field On Intracellular Calcium Concentration Of Rat Osteoblast Cell

ObjectiveTo investigate the proliferation of the static magnetic field of different intensities and action times on cultured osteoblast, more reach on the better intensity of the magnetic field to observe the free intracellular calcium ion concentration,it could help to discuss the possible mechanism of the static magnetic field on the proliferate function.Methods1.Cell culture:Isolated the calvaria from new bone rats,observed the shapes and structures of the cultured cells,the methods of alkaline phosphatase coloration and mineralized node staining were used to identify the osteoblast.2.The MTT assay:It was used to measure the rate of the proliferation on different magnetic field intensity(0Gs,260Gs,440Gs,900Gs)and action time(24h,48h,72h),every group adopt the mean value as the finally result.3. Fluorescence calcium measurement technique:The static magnetic field(440Gs) was chosen,the digitized video fluorescence microscope system and fluorescence double wavelength spectrophotometer were used to observe and measure the distribution and concentration of intracellular calcium ion. According to the fixed formula to account the intracellular calcium ion concentration,then get the mean value of each group.4.Statistic analysis:Statistical tests were performed by analysis of variance(ANOVA) and Dunnett-t test.Result1. The cultured osteoblast cell,adherence to the wall of fiask,and show the spindle,triangle or polyangular appearance. Alkaline phosphatase coloration and maineralized node stained by ARS.2.In the MTT assay,only the group 440Gs,osteoblasts appear significant proliferation within the 48h(P <0.01). When the static magnetic field effected on the cell at 72h,the group 44OGs and group 900Gs were rising significantly comparing with group 0Gs(P <0.01).3.Use the fluorescence dye to observe the intracellular calcium ion,it discovers that the [Ca²⁺]_i almost without change,signals of the fluorescence were relatively strong in the central lay,the strongest signal appears in the nucleus. And that the signal of calcium ion in the same lay was not equality.The [Ca²⁺]_i of osteoblast in group 15min was increased significantly under the static magnetic field(P>0.05).Conclusion1 .The static magnetic field could accelerate the proliferation of the cultured osteoblast,and may have certain intensity dependent.2.Certain intensity of static magnetic field could increase the rate of proliferation of the cultured osteoblast which may have time dependent.3.Certain intensity of static magnetic field had no instantaneous effect on the concentration of the intracellular calcium ion.4.Certain intensity of static magnetic field could increase the concentration of the intracellular calcium ion,and may have time dependent.