| Backgrounds:Research has indicated that after the menopause in adult women, the ovary function fails and the endogenous estrogen quality secreted reduces, the physics and chemical microenvironment in bone changes, it leads the balance of bone formation and bone absorption impaired, makes the loss exceed the formation of bone, leads to the result of the osteoporosis finally. It have verified already that the pulsed electrical magnetic field(PEMF) can accelerate the fracture union,make the delayed or disunioned fracture heal again through no operation. Since PEMF can promote osseous to form, then, whether PEMF has function of taking a turn for the worse of bone loss caused by menopause, it is the subject deserving to be probed into. Objectives:To investigate the effect and mechanism of PEMF on osteoblast and osteoclast co-cultured system , to probe into biology function of PEMF on bone metabolism of ovariectomized rat, to offer the experiment basis for clinical practice of PEMF for treating and preventing osteoporosis. Methods:1. To appraise the cultured osteoblast and osteoclast by using the morphology, peculiar enzyme staining and immune histolopathology technology.2. To determine the activity of osteoblast's ALPase after PEMF on osteoblast-osteoclast cocultured system; To probe into cell cycle and apoptosis of osteoblasts by FCM after PEMF;To analyse cell proliferation and the ablity of forming ore ization through image analytic technology after PEMF on cocultured system.3. To measure cell membrane electric potential and the density of ion of calcium in osteoblast through the fluorescence mark by FCM after dealing with PEMF in cocultured system.4. To analyse the change of histology of every group by methods of H-Estaining, tetracycline mark and type I collagen immune histolopathology; To determine the changes of bone microstructureses through image recombination technology.5. To detect the change of bone mineral density and biomethanics of every group.6. Statistical analytic methods: the software package of SPSS 10.0 is used for the test of significance. Mate t examine and variance analyse are used for measurable materials and LSD method among groups; square test is used for countable materials. Results:1. Osteoblast takes the form of long shuttleless and triangle type,itsticks wall to grow,.It shows large body, clear border, dark red afterbirth,the size and quantity of cell protruding vary, its nuclear locates in the edge of the cell, takes the form of round or oval, 1-2 kernels in cell is blue; the positive stain of type I collagen immune histolopathology and ALPase is the characteristic qualities of osteoblast, Thus we can appraise the cultured cell to be osteoblast.Osteoclast takes the form of multiple angles, It presents single nuclear forms in initial cultured stage, and then they merge step by step on later stage, there are a lot of small grains in the afterbirth. The cell membrane have more pseudopodium protruding styles. The positive stain of tartrit-resistant acid phosphatase (TRAP) and the ability of bone absorption are the characteristic qualities of osteoclast, Thus we can appraise the cultured cells to be osteoclast.2. The activity of ALPase arise after the dealing with PEMF.The cell cycle changs as the percentage of G 2+ S period increases, It indicate that synthesis of cell DNA and cell division increase. The osteoblasts proliferate largely and at the same time the apoptosis decrease.The ore ization of bone increase in osteoblast room and the function improves obviously.3. The membrane electric potential and the density of ion calcium of osteoblast is steady at sure base-line before PEMF process.( the intensity of fluorescence has not been changed), Behind PEMF process, cell membranepotential presents the depolarization; The density of ion of calcium rises in the cell. The function of membrane potential depolarization of PEMF can be blocked by TTX, the blocking calcium passway pharmaceutical Verapamil does not have obvious influence on it. But the effect of...
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