Abstract:Objective To investigate the effect of bone marrow mesenchymal stem cell transplantation transfected with nerve growth factor (NGF) gene overexpression vector on the repair of acute spinal cord injury in rats.Methods Human bone marrow mesenchymal stem cells (BMSCs) were extracted and the surface proteins of BMSCs were detected by flow cytometry.NGF overexpression vector plasmid was constructed by Cas-9 technology.BMSCs were transfected by lentivirus technology.The animal models of acute spinal cord injury in SD rats wereestablished by clamp method.SCI was constructed according to different treatment schemes.The models were divided into three groups: model group,BMSCs group and overexpression plasmid group.There were5models in each group.The NGF expression and nerve repair were detected by RT-PCR,Western blot and the immunofluorescence.Results The expression of CD44 and CD29 was 60.2% and 58.3% in the second generation BMSCs,while the expression of CD34 and CD45 was 3.4% and 2.6% in the second generation BMSCs by flow cytometry.The transfection efficiency of lentivirus was over 90%.The BBB score of the model group was the lowest one day after operation,and that of the overexpression plasmid group was higher than that of the model group and BMSCs group (P<0.05).Compared with model group and BMSCs group,the vacancy rate of hind limbs in overexpression plasmid group was significantly higher (P<0.05).RTPCR results showed that the relative expression level of NGF in overexpression plasmid group was significantly higher than that in model group and BMSCs group (P<0.05).The result of Western blot was the same as that of RT-PCR.Immunofluorescence results showed that NGF could co-stain with NeuN in the overexpressed plasmid group,and the number of neurons increased.The degree of spinal cord repair in overexpression plasmid group was better than that in BMSCs group and model group.It was found that the fracture spacing was obviously reduced and the repair of the broken end was better.Conclusion NGF overexpression plasmid can promote BMSCs to differentiate into neurons and promote the functional recovery of SCI animal model.
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