Study On Transplantation Of BMSCs By Three Different Ways To The Function Influence Of Spinal And Microglia After Spinal Cord Injury

Objectives:To isolate, culture and authenticate the BMSCs from the SD rats;To set up SCI models which by compressive clamp on thoracic10spinal cord. To investigate the function influence on microglia after spinal cord injury transplanting of BMSCs by local transplantation groups、tail vein transplantation groups and subara chnoid space transplantation groups.Methods:1、By whole bone marrow adherence method, we authenticate BMSCs through the test of potential ability of its, differentiation and surface antigens on CD29,CD34,CD45and CD90by flow cytometry.2. Establishing SD rats, SCI models by compressive clamp on thoracic10spinal cord, normal group;control groups (spinal cord injury6h group,12h group,24h group,72h group,1day group,3day group,7day group,14day group,21day group,28day group). BMSCs transplantations of spinal cord injury groups that redivided:local injection^tail vein injection and subarachnoid space injection.Every group was redivided,every group is5,a total of205. The treatment group was injected with the suspension of BMSCs in5hour after surgery. To observe nerve functions of hindlimb was evaluated by the BBB score in all groups. The cell morphology, distribution and quantitative change of microglia marked by OX-42were observed by immunohistochemistry. Inflammatory factor interleukin1β (TL-1β) in injured region were assayed by ELISA at6h,12h,24h and72h.Results:(1) The results showed that by whole bone marrow adherence method, BMSCs could be isolated and culture-expanded. BMSCs primary identification can be accomplished by flow cytometry.(2) SD rats’s SCI models were successfully set up.(3) BBB function score:one week late, BBB grading mark of control groups and of transplantation groups had no significant difference with that of normal groups (p>0.05). Compared with local transplantation groups and intravenous transplantation groups had no significant difference (p>0.05).(4) The number of OX-42positive cells are peak at the third day by immunohistochemisty staining. The number of OX-42positive cells at every observational time point in the treatment group and the injured group were outstandingly more than the sham surgery group (P<0.05),and the injured group showd the same condition compared with the treatment group(P<0.05). But the number of OX-42positive cells of subara chnoid space transplantation groups was lower than local transplantation groups and intravenous transplantation groups (p<0.05). The content of IL-1β on the6h,12h,24h and72h injured group were less than the control group (P<0.05).Conclusion:(1) BBB score showed:locomotor of function had being steady heighten in every transplantation groups.All three different transplantation ways have done to spinal cord injury and the subara chnoid space transplantation is the best.(2) BMSCs transplantation could work in depressing secretion of IL-1β and activity of microglia, and immunity inflammatory reaction. The subara chnoid space transplantation groups therapeutic efficacy is more obvious than local transplantation groups and intravenous transplantation groups.