| Objective:Investigate the changes on the expressions of FasL and caspase-3 in soleus muscle of damage side after transplantation of microencapsulated rabbit sciatic nerve cells/tissue treating spinal cord injury(SCI) in adult Sprague-Dawley (SD) rats. And the possible mechanism of the rehabilitation induced by the transplantation of the microencapsulated xeno-sciatic nerve cells/tissue treated SCI were discussed.Methods:The rabbit sciatic nerve cells/tissue suspension was prepared from 8 adult rabbits of either sex, weighting 3000g～3500g. A total of 80 adult Sprague Dawley rats (either sex, weighting 250g-325g) were experimented. Bilateral sciatic nerves pretreated from rabbits were dissected under aseptic condition and made into nerve cells/tissue suspension by trypsinization. The cells/tissue were intensively mixed with 1.5% sodium alginate solution, which was then extruded into 20 mmol/L barium chloride solution by using a droplet generation device, forming microencapsulated sciatic nerve tissue/cells suspension. All of the SD rats were randomly divided into four groups:8 in normal control group;24 in transplantation of microencapsulated xeno-sciatic nerve cells/tissue(microencapsulation group);24 in transplantation of xeno-sciatic nerve cells/tissue (experiment control group) and 24 in injury control group. Normal control group don't do any disposal.Microencapsulation group,experiment control group and injury control group were performed left hemisection of spinal cord at T10 level and subsequently transplanted with the gelatin sponge sticking 10μl microencapsulated nerve cells/tissue suspension,the gelatin sponge sticking 10μl cells/tissue suspension and gelatin sponge respectively. For Microencapsulation group,experiment control group and injury control group, In postoperative Id,3d,7d 14d, soleus muscle samples in damage sides were taken out respectively (each phase, take only 6 randomly).While corresponding samples were collected from 2 rats in normal control group each time. Hematoxylin-eosin (H-E) observes the muscle cells microstructure; Immunohistochemical method observes soleus apoptosis related proteins (FasL and Caspase-3) expressions. Results:H-E staining:normal group muscle structure is clear and aligned. Atrophy muscle in microencapsulation group is not obvious all the time, and the cytoplasm,nuclei of the muscle is relatively clear. In 3days, experiment control group and injury control group could see muscle atrophy,fracture; cells membrane incomplete; cells arrangement osteoporosis and disorder; cytoplasm light dying; nuclei hyperchromatic,size different,arranging disorderly; some cells appearing adipose vacuoles.7d-14d,atrophy and denaturation were more apparent. Immunohisto-chemical coloration:soleus in normal group disappear FasL or caspase-3 expression. In lday, soleus of experiment control group and injury control group occurred FasL protein and caspase-3 protein positive expressions, and gradually raised as time progress, in 7 days reached the peak, in 14days maintain at peak level. The expressions of FasL Protein and caspase-3 Protein in Microencapsulation group began to add in 1day,3days to highest,7days reducing,14days nearly down to normal level.And density(mean) of positive expressions of the microencapsulation group comparing with positive expressions of experiment control group and injury control group were significant different in each stage (P< 0.01);Experiment control group comparing with injury control group in different time wasn't statistically significant (P>0.05).Conclusion:transplantation of microencapsulated rabbit sciatic nerve cells/tissue in the treatment of spinal cord injury can inhibit expression of FasL and caspase-3 in soleus and delay muscular atrophy. |
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