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| Study on the repairing effect of BM-MSCs derived neuron-like cells on sciatic nerve injury in rats |
| HU Anquan FENG Qijun WANG Zheng’an QIN Li LI Zhe |
| Department of Orthopedics, Jiaxing First Hospital, Zhejiang Province, Jiaxing 314000, China |
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Abstract Objective To investigate the repairing effect of bone marrow mesenchymal stem cells (BM-MSCs) derived neuron-like cells on sciatic nerve injury rats. Methods BM-MSCs from 12 SD rats were isolated and purified, and they were divided into control group, β-mercaptoethanol group, all-trans retinoic acid group and Transwell cell co-culture group for seven days respectively. The cells with the highest induced differentiation rate were selected for animal experiments. A total of 90 rats were divided into sham-operation group, model group, general cell group and directional induction group according to the random number table method. The sciatic nerve crush injury model was established by clamping method, and 20 rats in each group were selected. The rats of general cell group and directional induction group were injected with BM-MSCs or neuron-like BM-MSCs respectively at the site of injury sciatic nerve. The rats in sham-operation group and model group were injected with the same amount of normal saline. The sciatic nerve function index (SFI), Bsso Beattie Bresnahan (BBB) scores and the recovery rate of muscle wet weight were compared among four groups. The expression of brain-derived neurotrophic factor (BDNF), myelin basic protein (MBP) and growth associated protein 43 (GAP-43) of nerve tissue were detected by Western blot. Results β-mercaptoethanol was used in subsequent animal experiments. SFI of model group at 2, 4, 8 and 12 weeks after operation were lower than those of the sham-operation group, and the differences were statistically significant (all P < 0.05). SFI of directional induction group at 2, 4, 8 and 12 weeks after operation were higher than those of model group and general cell group; SFI of general cell group at 2, 4, 8 and 12 weeks after operation were higher than those of model group, and the differences were statistically significant (all P < 0.05). BBB scores in model group at 2, 4, 8 and 12 weeks after operation were lower than those in sham-operation group, and the differences were statistically significant (all P < 0.05). BBB scores in directional induction group were higher than those in model group and general cell group at 8 and 12 weeks after operation; BBB scores of general cell group at 8 and 12 weeks after operation were higher than those of model group, and the differences were statistically significant (all P < 0.05). The recovery rate of muscle wet weight in model group was lower than that in sham-operation group, and the difference was statistically significant (P < 0.05). The recovery rate of muscle wet weight in directional induction group was higher than that in general cell group and model group; the recovery rate of muscle wet weight in general cell group was higher than that in model group, and the differences were statistically significant (all P < 0.05). The protein expression levels of BDNF, MBP and GAP-43 in model group were lower than those in sham-operation group, and the differences were statistically significant (all P < 0.05). The protein expression of BDNF, MBP and GAP-43 in directional induction group were higher than those in model group and normal cell group; and the protein expressions of BDNF, MBP and GAP-43 in general cell group were higher than those in model group, and the differences were statistically significant (all P < 0.05). Conclusion β-mercaptoethanol can induce BM-MSCs to differentiate into neuron-like cells to obtain the stable neurons. In addition, neuron-like cells directed by mesenchymal stem cells has obvious repair effect on sciatic nerve injury.
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