| Objectives: (1) To isolate, culture and authenticate the BMSCs from the SD rats. (2) To investigate the effect on microglia activity and inflammatory reaction with BMSCs early injected into SD rats,SCI models which were set up by compressive clamp on thoracic 10 spinal cord through caudal veins.Methods:(1) BMSCs were separated from SD rats,tibias and femurs by whole bone marrow adherence method .We authenticate BMSCs through the test of potential ability of its,differentiation and surface antigens on CD29,CD34,CD45 and CD90 by flow cytometry. (2) Establishing SD rats,SCI models by compressive clamp on thoracic 10 spinal cord. The SD rats were randomly divided into the sham operation group, control group and treatment group. The treatment group was injected with the suspension of BMSCs and the suspension of non-serum L-DMEM on the control group and sham operation group by way of caudal veins injection in 5h after surgery. The cell morphology,distribution and quantitative change of microglia marked by OX-42 were observed at the 1d, 3d, 7d, 14d and 21d by immunohistochemistry;nerve functions of hindlimb was evaluated by the BBB score at 1d,3d,7d ,14dand 21d in all groups. Inflammatory factor interleukin 1β(IL-1β) in injured region were assayed by ELISA at 6h,12h,24h and 72h.Results: (1) BMSCs cultured in vitro grew in clone-type with high proliferation rate. The flow cytometry showed that BMSCs express CD29 and CD90,but do not express CD34 and CD45.They can be induced into neuroblasts, osteoblastses and lipoblasts in special condition. (2) SD rats,SCI models were successfully set up. The number of OX-42 positive cells at every observational time point in the treatment group and the injured group were outstandingly more than the sham surgery group (P<0.5),and the injured group showd the same condition compared with the treatment group(P<0.5). The BBB score of the injured group and the treatment group were lower than the sham surgery group (P<0.05), and there no significant difference between the1st day,3rd day,7th day the treatment group and the injured group (P>0.05),but 14th day,21th day of the two groups have big differenee(P<0.05).The content of IL-1βon the 6h,12h,24h and 72h injured group were more than the sham surgery group and the treatment group (P<0.5),while in the 6h,12h and 24h the treatment group showed outstandingly difference compared with the sham surgery groupConclusion: (1) The whole bone marrow adherence method is a useful, safe, and efficient way to isolating and culturing BMSCs from SD rats;(2) Early intravenous injection with BMSCs could work in depressing secretion of IL-1βand activity of microglia, and promote functional recovery.
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