Influence Of Intravenous Administration Of Bone Marrow Mesenchymal Stem Cells On Expression Of VEGF,bFGF And Regeneration Of Blood Vessel After Spinal Cord Injury In Rats

Objective:To observe the survival and migration of Bone Marrow Mesenchymal Stem Cells labeled and the change on expression of VFGF and bfgf factors at different times after transplantion MSC via caudal vien to treat contused spinal cord injuried adult SD rats model, and to provide experimental evidence for the reparing function and menchanism which MSCs treat spinal cord injury.Methods:1.Rat bone marrow mesenchymal stem cells(MSCs)from adult rats were cultured in vitro,their specific cellular surface marker were analysed with flow cytometer.2.Experimental animal and groups:42 healthy adult srague-dawely(SD)famal rats,mean weigh 200-250g,were randomly divided into 7 groups including normal control group,control group and MSCs transplantation groups,each groups 6 rats.3.Establish contused spinal cord injuried SD rat model: To adapt the Allen's improved method-20g weight iron bar fall from 5cm high and strike at T12 segment spinal cord to get contused spinal cord injuried rat models.4. MSC are prelabeled with CFSE in vitro before transplantation.5. Transplantation of MSCs:Inject MSCs into the spinal cord injury site after injury 3days in transplant groups,but PBS are injuried in contused control groups.6. Materials and Estimates6.1 Heart perfusion,draw the spinal cord from rat,slice6.2 The survive of CFSE-MSCs which were transplanted into spinal cord injuried rats via vein was observed by using confocal microscopy6.3 The expression of VEGFand bFGFmRNA was detected in spinal cord by using RT-PCR.6.4 Immunohistochemistry identification of VIII-Ag expression6.5 Functional evaluation using the open-field BBB scoring system and climd grid training.7. statistical analysisResult: 1. CFSE-MSCs which were transplanted into spinal cord injuried rats via vein could survival in the spinal cord by using confocal microscopy ,show the ability of secreting VEGF,bFGF.2. RT-PCR revealed that the expression of VEGFand bFGFmRNA was higher in the MSCs transplantation group than that of the control group on the 3th,14th day and the counted number of VEGFand bFGFmRNA increase obviously in the MSCs transplantation group. But MSCs transplantation group compared with control group on 1th day(P>0.05 ) , the expression of VEGFand bFGFmRNA was not statistical significance.3. Immunohistochemistry revealed that the expression of MVD was higher in the MSCs transplantation group than that of the control group on the 3th,14th day and the counted number of MVD increase obviously in the MSCs transplantation group. But MSCs transplantation group compared with control group on 1th day(P>0.05) , the expression of VEGFand bFGFmRNA was not statistical significance.4. The result of BBB scoring system and climd grid training indicated that MSCs could improve the recovery of motor funcyion of spinal cord injuried rat model.Conclusion: MSCs,after being implanted into injured spinal cord by intravenous administration ,shows the ability of secreting VEGF,bFGF,with subsequent angiogenesis.