| 0bjective1 To preliminary investigate the influence on the brain derived neurotrophic factor (BDNF)expression in the brain of traumatic brain injury (TBI) rat after transplanting bone marrowmesenchymal stem cells (BMSCs) and vascular endothelial growth factor (VEGF).2 To preliminary study on combined transplantation of BMSCs and VEGF for treatment of TBIrats and its therapeutic effect.3 To establish the experimental base for further study of BMSCs transplantation in treatment ofTBI.Methods1 40 SD rats were randomly divided into 4 groups: individually transplanting BMSCs group,individually transplanting VEGF group, transplanting BMSCs and VEGF group and the controlgroup, 10 rats in each group. Moderate traumatic brain injury model was constructed bythe improved Feeny free-falling impact method.2 Femoral bone marrows were obtained from 2 SD rats and cultured in vitro by the adherentenrichment method.3 After modeling for 1 week, the same amount of BMSCs, VEGF, BMSCs plus VEGF and PBSfluid are respectively injected into the corresponding rat brain tissue surrounding the injury instereotactic instrument.4 2 weeks later, the rats were put to death and the brain were obtained. Using HE and Nisslstaining to observe the pathological changes of brain tissue; With immunohistochemical staining,using Image pro-Plus6.0 software to measure the sum cumulative optical density of the BDNFpositive expression regions in the penumbra and the uninvolved symmetric parts. The mean ofsum IOD of 10 consecutive horizons was used to represent the BDNF content of the part.5 The means of sum IOD are measurement datas, with the mean±SD（±S）said. The sumIOD of penumbras was compared with single factor analysis of variance between theexperimental group and the control group. The mean of sum IOD of symmetric part and thepenumbra was compared with the paired t test. The diffierence has statistical significance with P< 0.05. SPSS13.0 statistical software was applied for processing.Results1 Establishment of rats moderate TBI model: 3 rats rapidly dead after being impacted. 4 ratsdead 8 hours later. Other rats were randomly selected and impacted to replace the deads. Finally, 40 TBI models were successfully established.2 Isolation and culture of rat BMSCs: The isolated and cultured cells had the BMSCs’biologicalcharacteristics: adherent growth, spindle-shaped. Primary BMSCs appeared after 7-day longculturing.3 Stereotaxic transplantation：Rats in each group were all successful transplanted, nopostoperative death.4 Pathological examination: HE staining showed that interstitial was dense, the neuronalmorphology was normal in the symmetry. Penumbra interstitial slightly loose, the density ofneurons was slightly lower than symmetrical part, neurons had edema, shrunken neurons wereoccasionally visible. Spindle BMSCs were visible in the granule cell layer and penumbras ofgroup BMSCs and group combined transplantation. Nissl staining showed Nissl bodies in thepenumbra were slightly less than symmetric part. Immunohistochemical staining showed thatpositive cells were widely distributed in the brains, and BDNF was predominantly expressed inneurons, microglia, astrocytes and bone marrow mesenchymal stem cells.5 Except the BMSCs group, the BDNF contents of symmetrical parts were higher than thecontents of penumbras in other groups (P < 0.05); The BDNF contents of the penumbras incombined transplantation group was significantly higher than the contents in control group (P <0.05).Conclusion1. Combined transplantation of BMSCs and VEGF can effectively inhibit the decrease of BDNFcontents of TBI rats’penumbras.2. The therapeutic effect of combined transplantation of BMSCs and VEGF for the treatment ofTBI may better than transplantation of BMSCs. |
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