Influence of the pulsed electric magnetic field on cell membrane potential and calcium ions of osteoblast in the osteoblast osteoclast co-culture system
Jian Ting Chen,Xiao Yun Tan,Chun Lu Yang,Huan Zhang Tang,Da Di Jin Jian Ting Chen,Xiao Yun Tan,Chun Lu Yang,Huan Zhang Tang,Da Di Jin,Department of Spine and Osteopathy,Nanfang Hospital,First Military Medical University,Guangzhou 510515,Guangdong Province,China
AIM:To investigate the effects of pulsed electric magnetic fields(PEMF) on cell membrane potential and calcium ions of osteoblast in the osteoblast osteoclast co culture system in order to find the theoretical foundation for PEMF in the treatment of the osteoporosis. METHODS:The osteoblasts and osteoclasts were isolated from the SD mice of one day and co cultured in the same system in vitro.All the co cultured systems were divided into four groups:control group,verapamil,verapamil(Ver) group,tetrodotoxin(TTX) group and Ver+TTX group.The membrane potential of osteoblasts was performed,and calcium ions were stained in each group according to fluorescence probe DiBAC4(3) and Fluo 3/AM combininmg with living cell membranes and calcium ion peculiarly,and then flow cytometry(FCM) technology was utilized to measure the changes of fluorescence intensity pre and post PEMF so as to reflect the cell membrane potential and the concentration of cell calcium ions indirectly. RESULTS:The cell membrane potential and the concentration of calcium ion were steady in certain base line level before treated with PEMF in each group(the control group:37.68±1.81 and 164.24±2.82 respectively).After treatment with PEMF,the fluorescence value of the cell membrane potential began to rise after 40 s in the blank group,and reached the peak value after 150 s(75.05±2.27). About 60 s later,the fluorescence value of calcium rised,and reached the peak value after 120 s(237.22±2.67),after that it was steady in certain level all the time in the course of PEMF process.The depolarization effect of PEMF was blocked by TTX,but not by Ver;But both TTX and Ver could inhibit the effect of rising the concentration of calcium ion of PEMF. CONCLUSION:PEMF causes cell membrane potential to depolarize,and then calcium ion passage opens and calcium ions pour into cells.As the second messenger of the cell,calcium ion mediate the regulation of PEMF to the osteoblast function.
【CateGory Index】： R454.1
【CateGory Index】： R454.1