The Therapeutic Efficacy Of Lentiviral Vector Mediated BDNF Gene-modified RMSCs In Sciatic Nerve Injury

Objectives Lentiviral vector carrying the BDNF gene infects the rat mesenchymal stem cells (rMSCs) to get BDNF gene-modified rMSCs.Methods The lentiviral plasmid PNL-BDNF- IRES2-EGFP, with the packaging plasmid pHELPER and the envelop plasmid pVSVG were co-transfected to 293T cells to produce BDNF gene and the reporter gene enhanced green fluorescent protein (EGFP) gene co-expressing lentivirus. rMSCs were separated from the bone marrow of two-month -old F344 rats, cultured in vitro, and identified. The rMSCs were infected by the LV vectors that were produced already and were identified with fluorescent microscope, immunocytochemical staining。Results After three plasmids of lentiviral vector co-transfecting to the 293T cells with the fluorescent microscope ,a lot of green fluorescence in 293T cells was observed. The supernatant was concentrated by ultracentrifugation and the titer of lentivirus measured was 6.5×107 TU/mL. After the constructed lentivirus infected rMSCs the green fluorescence and BDNF expression in rMSCs were observed.Conclusions BDNF gene-modified rMSCs could express BDNF abundantly. This facilitates our next study on injury sciatic nerve in rats.Objectives To observe the therapeutic efficacy of lentiviral vector mediated BDNF gene modified MSCs in sciatic nerve injury.Methods : The right sciatic nerves of 60 adult F344 rats were crushed with forceps, these rats served as the experimental animal models, then they were randomly divided into PBS group, rMSCs group and BDNF-rMSCs group, and 2μL BPS, 2μL MSC solution and 2μL BDNF-MSC solution were transfered into injury site of each group with microinjection respectively.Result: The grafted MSCs were survival at the injury site in the rMSCs group and BDNF-rMSCs group. Moreover, the grafted rMSCs expressed BDNF abundantly in BDNF-rMSCs group. Compared with PBS group and rMSCs group, the number of HRP positive cells in the Lumbar 4,5 spinal cord were increased (P<0.01 ) and the recovery of nervous functional defects was better(P<0.01) in BDNF-rMSCs group.Conclusion: Transplantation of BDNF gene-modified rMSCs is an efficient method for the treatment of peripheral nerve injury.