| Objective:To investigate the therapeutic effect of ultrashort wave(USW)combined with bone marrow mesenchymal stem cells(BMSCs)on spinal cord injury(SCI)in rats.Methods:BMSC.s were cultured primary culture to the third generation by adherent culture method,and the third generation BMSCs surface antigens CD45 and CD90 were detected by immunofluorescence assay.The spinal cord injury model was established by modified Allen's method and randomly divided into Five experimental groups:sham group,control group,BMSCs group,USW group and BMSCs + USW group;Each group divided into four subgroups.there were 2 rats in each subgroup of sham group,other subgroups contained 4 rats.BBB scores were evaluated before and after spinal cord injury,Id,7d,14d and 28d after treatment,respectively.The rats were sacrificed within 4 weeks after treatment.The expression of inflammatory cells,iNOS and Arg-1 in the spinal cord were respectively detected by HE staining,immunohistochemistry and Western blot at 1d,7d,14d and 28d after SCI.Results:The primary and third generation of BMSCs were successfully cultured,and the expression of CD45 and CD90 on cell surface was detected by immunofluorescence.The BBB score of sham operation group before and after operation was unchanged.At the first day after SCI,the score of BBB score was the lowest among the four groups,and there was no significant difference among the four injury groups(P>0.05),and with the extension of time,the motor function of all injury groups showed different degrees of recovery.From the first week after SCI,BMSCs group,USW group and BMSCs + USW group(All P<0.001),Compared with the control group,the BBB score showed an increasing trend in the 2-4 weeks after the treatment,showing significant differences(all P<0.001);While the BBB score of the two groups(USW group\the BMSCs + USW group)showed no significant difference in the first two weeks after SCI(P>0.05),but at the fourth week,the comparison between the two groups started(P<0.01).However,there was a significant difference at the first week between the USW group and the BMSCs +USW group(P<0.05,P<0.05).From second week to fourth week after SCI,the differences continue to increase(all P<0.01);Therefore,in improving the behavioral function of rats,it is based on the role of USW,BMSCs play a supporting role.HE staining showed that inflammatory cells in BMSCs + USW group,USW group and BMSCs group were significantly less than those in control group one week after injury,and the area of necrotic cavity was relatively reduced;Immunohisto-chemistry results shown that the expression of iNOS in Injured spinal segments is raised,and reach the peak in the second week after SCI.In the second week after SCI,the expression of iNOS positive cells in BMSCs group,USW group and BMSCs +USW group were both lower than that in Contral group(All P<0.001).Compared with the USW group,the expression of iNOS positive cells in BMSCs group was less,but there was no significant difference(P>0.05).BMSCs + USW group,the expression of iNOS positive cells was the lowest,which was lower than that of the BMSCs group and the USW group(all P<0.001).At the 4th week,the expression of iNOS between the BMSCs group and the USW group still no difference(P>0.05).However,the expression of iNOS in BMSCs + USW group compared with BMSCs group and USW group has significance difference(P<0.05,P<0.001).The expression of Arg-1 was increased And peaked at the first week.The number of Arg-1 in BMSCs + USW group,BMSCs group and USW group in the first week increased compared with the control group,which was significant(P<0.001).The positive expression of Arg-1 in BMSCs group was lower than that in USW group,but there was no significant difference(P>0.05).The positive expression of Arg-1 in BMSCs + USW group is higher than that in BMSCs group and USW group,and difference is significant(P<0.001,P<0.01).The expression of Arg-1 in BMSCs +USW group,BMSCs group and USW group was still higher than that in control group at the 4th week,the difference was statistically significant(P<0.001).However,the expression of Arg-1 in each group was lower than that in the first week,there was no significant difference between the USW group and the BMSCs group(P>0.05),while the Western blot results showed that the BMSCs + USW group,the expression of iNOS in the first week after spinal cord injury was less than that in the other three groups.However,the expression of Arg-1 in the BMSCs + USW group was increased relative to the other three injury groups in the second week after spinal cord injury.Conclusion:The combination of ultrashort wave(USW)and bone marrow mesenchymal stem cells(BMSCs)can repair and treat spinal cord injury in rats. |
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