| Object:To explore the effect of BMP-7 gene transfection into bone marrow mesenchymal stem cells(BMSCs)transplantation to treat spinal cord injury in rats.Methods:The density gradient centrifugation method combined with the whole bone marrow adherence method was used to separate and culture BMSCs.After cultured in vitro to the third generation,the cell surface markers CD44 and CD29 were detected by flow cytometry.Adipogenic induction and osteogenic induction test whether the cells have the ability of stem cell differentiation.BMSCs were transfected with LV-BMP7-GFP conjugate to evaluate the expression of green fluorescent protein GFP.After modeling,they were randomly divided into sham operation group(group A),control group(group B),BMSCs group(group C),and LV-BMP7-BMSCs group(group D).Treatment was performed on the 3rd day after the operation.After 1d,3d,7d,14 d,21d,28 d after treatment,the BBB score standard was used to evaluate the recovery of the motor function of the lower limbs of the rats after treatment.After 28 days of treatment,immunohistochemistry and immunofluorescence were used to detect the expression of NF-200 and GFAP.After 28 days of treatment,horseradish peroxidase retrograde tracing method was used to detect the remodeling of neuronal circuits.Results:On the 14 th,21st,and 28 th day after spinal cord injury treatment,the BBB score of group D was higher than that of group B and group C(P<0.05).The expression level of NF200 in group D was higher than that in group A,group B,and group C by immunohistochemistry(P<0.05).Compared with group A,the expression level of GFAP in group B,group B,and group D increased(P<0.05).Comparing the three groups,there was no difference in the expression level of GFAP(P>0.05).The percentage of red fluorescent area of NF-200 in group D was higher than that in group A,group B and group C by immunofluorescence(P<0.05).Compared with group A,the percentage of GFAP green fluorescent area in group B,C and D is higher(P<0.05),but there is no statistical difference in the percentage of GFAP green fluorescent area between group B,C and D(P>0.05).The number of nerve cells positively labeled by horseradish peroxidase in group D was higher than that in groups B and C(P<0.05).Conclusion:BMP-7 gene transfection of BMSCs and transplantation to treat rat spinal cord injury can promote the repair of spinal cord injury neurons and remodeling of neuronal circuits,which is beneficial to the recovery of nerve function after spinal cord injury in rats.It is the ultimate application of BMSCs transplantation for the treatment of SCI Provide a theoretical basis for the clinic. |
