The Role Of Olfactory Ensheathing Cells (OECs) In Axon Regeneration Of Rat After Spinal Cord Injury (SCI)

Objectivespinal cord injury often leads to a poor ending and burdens families of the patients great much. So it causes great concern. One of the key step to restore normal function is the enhancement of the axonal regeneration. While the transplantation of OECs is proved to be a relative satisfactory method to encourage ax-ons to regenerate. Although OECs have great advantages in promoting axon regeneration and reformation of sheath, the method has weakpoints, such as, whether OECs have definite ability to provoke regeneration and to what extend this ability is, how to maximize this ability. The current research is aiming at the observation of the biological characteristics of OECs cultured in vitro and their actions on the artificial spinal cord injury of the rats.Methodsculture, purification and identification of the OECsCulture and purify the OECs by the modified Nash's method,then identify with P75 antibody using SP staining In vitro.The establishment of the SCI model and the transplantation by mirco - injection.The adult Wistar rats were randomly separated into two groups, spinal cord was exposed under apepsis and the left half was sectioned. Suspension of cultured OECs or DF - 10S culture media were stereotaxically mirco - injected into the ends of the incision .Motor functional evaluation by BBB scaleBy the end of 1 day,. 3 day ,1 week,2 week,3 week and 4 week after injury, the hind limb motor function were evaluated by BBB scale. The data were analyzed statistically with nonparametric test.HE stainingThe slides were cut longitudinally for HE staining and observed for the scarImmunohistochemistry stainingThe slides were cut longitudinally , stained by SP method with P75, NF200 and GAP -43 and observed for OECs surviving and axon regenerationWesternblot detectionThe axpression of NF200 and GAP - 43 were detected by alkaline phospha-tase staining.Resultsculture , purification and identification of the OECsOECs grew regularly, the more longer they grew, the more regularly they grew. The cell membrane of positive OECs show brown, and cell nucleus show light blue; as for the negative cells, only the cell membrane show light blue.Motor functional evaluation by BBB scaleThere were no difference between two groups within the first week,and the scores of experiment was higher then the control group after one week, but there was significant difference only by the end of the 4th week( P <0.05).HE stainingThere was glial scar existing in the injury region, while the uninjured spinal cord still kept the norm structure. Immunohistochemistry stainingP75 positive cells mainly dispersed in the glial scar,some distributed linearly. NF200 and GAP -43 positive fibers mostly distributed in the uninjury region and a part of GAP -43 positive fibers entered into the scar, the front end of NF200 and GAP - 43 positive fibers turned to injury region in experiment group, but not in the control group.Westernblot detection...