| Objective To investigate effects of transplantation of bone marrow stromal stem cells (BMSCs) combined with granulocyte colony stimulating factor (G-CSF) on repair of the acute spinal cord injury (SCI) in adult rat.Methods BMSCs were separated and cultured from four Wistar rats aged 8 weeks.24h before transplanting, BMSCs of the third generation were labeled by Hoechst 33342, then adjusted the labeled BMSCs to 1×105, prepared monoplast suspension for use.48 adult wistar rats were made the acute spinal cord injury (SCI) models at T11 level according to the improved Allen's method and the requirement of experiment, the qualified animals were randomly divided into four groups(n=12):BMSCs and G-CSF treatment group (group A), BMSCs transplantation group (group B),G-CSF treatment group (group C), single SCI group (group D).After 1,2,3 and 4 weeks of surgery, the functional recovery of the hind limbs was evaluated by the Basso-Beattie-Bresnahan (BBB) locomotor rating score.after 4 weeks of surgery, Specimens were procured at 4 weeks after surgery for investigating the survival and differentiation of BMSCs by fluorescence microscope in the damaged area and border area. In addition, gross anatomy and histology were observed by HE staining.Immunofluorescence measurement were used to detected expression of neuron-specific enolase (NSE),glial fibrillary acidic protein (GFAP) and neurofilament 200 (NF-200) proteins.Results 1-4 weeks after surgery, the values of BBB in group A continually increased,the BBB scores in group A were significantly higher than other groups, Group D was the lowest, the differences were statistically significant (P<0.01);B Group higher than Group C at 3,4 weeks (P <0.01), Group D performed significant difference with group B and group C (P< 0.01).HE staining revealed that BMSCs combined with G-CSF on repair of spinal cord injury were best, Group D were the worst, Group B,C ranged between A and D groups.4 weeks after surgery, Immunofluorescent staining results showed there were a great quantity of BMSCs gathered which were labeled by Hoechst 33342 in the SCI damaged area and border area in gray matter more than white matter, migrated from the transplantation spot to the injured tissue,The number of survival cell in group A was more than group B.The NSE,GFAP and NF 200-positive cells around the SCI in the group A were higher than group B. The NSE, NF 200-positive cells were no obvious, but GFAP-positive cells were observed in the lesioned area in group C, and extended to the cavity. The damaged around of SCI in group D were filled with a large number of GFAP-positive cells of Structural mess,regeneration and NSE or NF-200 positive cells were no significant.Conclusion Transplantation of BMSCs could survived and differentiated; BMSCs and G-SCF.both combination can promote structure repair and nerve function recovery. There may be a synergistic effect.
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