The Effect Of Curcumin On Prevention Of Epidural Scar Adhesion After Laminectomy

Objective:Use the rat animal experiment to investigate the effect of local application of curcumin on prevention of scar adhesion after laminectomy&find out the specific mechanism of preventing epidural scar adhesion by cell experiment.Method:Thirty-six male Wistar rats were randomly divided into 3 groups:50 mg/ml of curcumin group,100 mg/mL of curcumin group and saline control group.Disinfect the lumbar spine surgery and do skin preparation.Separating tissue layer by layer at the lumbar vertebrae of L1-L2.The L1-L2 lumbar vertebrae were removed to form a 0.2 cm×1.0 cm defect,exposing the dura mater,and suture the erector spinae later.Different concentrations of curcumin and normal saline were injected into the epidural space.Intramuscular injection of penicillin into rats for 3 days to prevent wound infection.One month later,6 rats were randomly selected from each group,using the Rydell-balazs standard to evaluate the degree of adhesion in the scar.Fixing the rat tissue with formaldehyde,the specimens were embedded in paraffin.HE staining was used to observe the effect of curcumin on epidural scar adhesion in rats under 40 times microscope,and observe the number of fibroblasts in the scar under a 200-fold microscope.Masson-stained sections were observed under a 200-fold microscope to detect the content of collagen fibers in scars.Culturing human fibroblasts in vitro,and the fibroblasts were treated with curcumin at a concentration gradient(0 ?.M.5 ?M,10?M,20 ?M,40 80 ?M)for 24 h,detecting the cell viability of fibroblast by CCK-8.Fibroblasts were treated with curcumin at a concentration gradient(0 20 ?M,40?M)for 24 h,using EdU proliferation staining method to further confirm that curcumin has an inhibitory effect on fibroblast proliferation.Fibroblasts were treated with curcumin at a concentration gradient(O?M,10?M,20?M,40?M)for 24 h,Western blot was used to detect the expression of Wnt3a.?-catenin,GSK-3? in Wnt/?-catenin pathway and the expression of proliferation-related protein CyclinD1 and PCNA.Results:The specimens showed that the curcumin group had less scar adhesion and scar area than the control group,and the high concentration group had better effect than the low concentration group.HE staining showed that compared with the control group,the scar density and adhesion range in the epidural area of the curcumin treatment group were smaller than those in the control group,and the number of scar tissue fibroblasts was also less than the control group.Masson staining showed that the collagen content of the curcumin-treated group was reduced.The results of CCK-8 showed that curcumin can reduce the cell viability of fibroblasts.As the concentration of curcumin increases within a certain range,the cell viability of fibroblasts gradually reduces.The results of EdU showed that curcumin reduced fibroblast proliferation,and the rate of EdU-labeled positive cells in the curcumin-treated group was lower than that of the control group.Western blot analysis showed that curcumin could decrease the expression of Wnt3a,?-catenin,CyclinD1 and PCNA,and increase the expression of GSK-3?.With the increase of curcumin concentration in a certain range,the effect of this regulation is more obvious.All the results indicate that curcumin can inhibit the expression of Wnt/?-catenin signaling pathway to inhibit the proliferation of fibroblasts,and to prevent epidural scar adhesion after laminectomy in rats.Conclusion:Curcumin can be used to prevent epidural scar adhesion after laminectomy.The underlying mechanism may be to inhibit fibroblast proliferation by down-regulating Wnt3a/?-catenin pathway expression.