| OBJECTIVE: To make further efforts to investigate the effects of low frequency electromagnetic fields(LFEMF) on promoting the induction of bone marrow-derived neural progenitor cells(BM-NPCs)into nerve cells and the m RNA expressions ofβ-IIItubulin METHODS: Firstly, The BM-NPCs were cultured and induced under neural culture conditions. Secondly, the BM-NPCs were treated with a sine wave electromagnetic fields,the frequency of 50 Hz,and magnetic induction of 5 m T, and 60 min per day for fifteen days. And the control group was setted up at the same time. After BM-NPCs induced differentiation, the cell markers of Nestin,Tuj-1 and GFAP were analyzed by Immuneflurorescent and the m RNA expressions of Nestin, PSA-NCAM, β-IIItubulin,ACHE, GABA and5-HT were tested by real-time quantitative reverse transcriptasepolymerase chain reaction method. Neuron-like cell electrophysiology was tested by neurophysiological testing RESULTS: 1, After culturing in serum-free neural sterm cells suspension culture conditions,the BMSCs could be aggregated into BM-NPCs that were positive for nestin. The results of immunofluorescence after the induction of BM-NPCs indicated that the induced neuron-like cells which positively expressed Tuj-1 and GFAP in two groups. The result of Q-PCR showed that the m RNA expressions of Nestin, PSA-NCAM, β-IIItubulin, ACHE, GABA in two goup were statisticallydifferent from that befor BM-NPCs induction in vitro(P<0.01).The β-IIItubulin m RNA level in the electromagnetic field group was higher than in BM-NPCs(P<0.05). Cell electrophysiology showed a spontaneous postsynaptic currents in the cell,with a 60 m V current amplitude and 200±30 p A(mean±SEM,n=6) current intensity.CONCLUSION: The low frequency electromagnetic fields can facilitate the induction of neural progenitor cells to differentiate into nerve cells,it may apply to up-regulated the m RNA expressions of β-IIItubulinOBJECTIVE: To investigate the effects of LFEMF on locomotor functions of traumatic brain injured rats after BM-NPCs transplantation therapy.METHODS: Traumatic brain injured rat models were established. Then the BM-NPCs labeled with CD-Dil were transplanted into the centre part of injured brain tissues. The rats were divided into control group(C group), EMF group( E group),BM-NPCs group(B group) and EMF+BM-NPCs group(EB group). E group and EB group were exposed to a sine wave electromagnetic fields 60 min per day for sixty days,with the frequency of 50 Hz, and magnetic induction of 5 m T. Locomotor function were assessed by the Wayne Clark scale and the grooming deficit score on 1st, 3rd, 7th, 30 th,60th days after transplanting BM-NPCs. On 1st, 7th, 30 th days after transplanting BM-NPCs, rats in each group were sacrificed respectively to gain brain tissues sample.The pathology of injured brain tissues in each group were tested with HE staining.Survival condition of the BM-NPCs labeled with CD-Dil was observed and Neu N, GFAP,β-IIItubulin postitive expression of BM-NPCs in injured brain tissues were tested by immunofluorescence.RESULTS: 1. Behavioral score: On 1st day after transplantating, the Wayne Clark scale and the grooming scale of each group was not statistically significant(P>0.05), While on 3rd, 7th, 30 th, 60 th days, the Wayne Clark score and the grooming score of four groups compared with each other were different statistically(P<0.05), with the EB group changed most significantly. 2. HE staining: On 7th day after establishing traumatic brain injured rat models, the injured brain structure was damaged, andthe vascular was compressed and deformative, and the blood flow was further reduced,.The nerve cells in each group were swelling, necrosis and degenerated.On 7th day after transplantating, tissue structure in C group and E group was swelling, and the cystic cavity and the nerve cells were obveriously decreased, surrounded with inflammatory cells, but the cystic cavity area were limited, and swelling were alleviate and glial cells can be seen in B group and EB group.. On 30 th day after transplantating,the brain organizational structure of each group was gradually improving, except C group. The cystic cavity area of other groups became smaller.Swelling tissues and inflammatory cells were disappearing and the cells were arranged orderly in EB group. 3, On 30 th day, we could observe a large amount of GFAP positive cells around the injured brain area in C group, but GFAP protein in B group and EB group were less than C group. On 7th ady, Neu N and β-IIItubulin positive cells labeled with CD-Dil were not seen around the injured brain area in B group and EB group. On 30 th day, the Neu N andβ-IIItubulin positive cells labeled with CD-Dil could were connected with the damage area in B group and EB group,with EB group obviously.CONCLUSION: Low frequency electromagnetic fields could improve the side lims locomotor function of rats with traumatic brain injured, that may related with the LFEMF can facilitate the induction of the bone marrow stroma cells-derived neural progenitor cells that was implanted on rats with traumatic brain injured to differentiate into neurons... |
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