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2022, Vol. 26 ›› Issue (16): 2551-2556Topical application of mycophenolate mofetil prevents epidural fibrosis by inhibiting the proliferation and migration of fibroblastsZhu Dongming1, Zhang Zhen1, Zhang Jie2, Yan Lianqi3
Abstract: BACKGROUND: Studies have shown that epidural fibrosis after laminectomy is mainly caused by the proliferation and migration of fibroblasts, and mycophenolate mofetil can inhibit the proliferation and migration of fibroblasts. OBJECTIVE: To explore the effect of topical application of mycophenolate mofetil in preventing epidural fibrosis after laminectomy and its possible mechanism. METHODS: (1) Cell test: Fibroblasts were separately treated with 0, 0.01, 0.1, 1, 10, and 100 μmol/L mycophenolate mofetil solution for 24 hours. Cell viability was detected by CCK-8 assay, and the appropriate treatment concentration was selected for subsequent cell experiments. Fibroblasts were separately treated with 0, 0.1, 1, 10 μmol/L mycophenolate mofetil solution for 24 hours. EdU assay and cell cycle assay were used to detect cell proliferation. Wound scratch assay and transwell assay were used to detect cell migration. Western blot assay was used to detect the expression of cell proliferation-related proteins (nuclear proliferation antigen, Cyclin D1) and migration-related proteins (α-tubulin, vinculin). (2) Animal test: Forty-eight adult SD rats were selected to construct rat laminectomy model and randomly divided into four groups. In the control group, cotton pads soaked with normal saline were placed on the postoperative bone defect area. In the low, medium, and high concentration groups, cotton pads soaked with 2.5, 5, and 10 g/L mycophenolate mofetil solution were placed on the postoperative bone defect area. Four weeks after the operation, the vertebral body of the operative area was taken for histological analysis. RESULTS AND CONCLUSION: (1) Cell test results: CCK-8 assay showed that mycophenolate mofetil inhibited the viability of fibroblasts in a concentration-dependent manner. EdU assay showed that mycophenolate mofetil inhibited the proliferation of fibroblasts in a concentration-dependent manner. Flow cytometry showed that 0.1 μmol/L mycophenolate mofetil solution blocked the cell cycle in G0/G1 phase. Wound scratch assay and transwell assay showed that mycophenolate mofetil inhibited the migration of fibroblasts in a concentration-dependent manner. Western blot assay showed that mycophenolate mofetil inhibited the expression of fibroblast proliferation-related protein and migration-related protein in a concentration-dependent manner. (2) Animal test results: Histological observation showed that with the increase of mycophenolate mofetil concentration, the number of fibroblasts and collagen production in the fibrotic tissue of the surgical area gradually decreased. (3) The results showed that the topical application of mycophenolate mofetil after laminectomy can effectively prevent the occurrence of epidural fibrosis, possibly by inhibiting the proliferation and migration of fibroblasts. Key words: material, mycophenolate mofetil, fibroblast, proliferation, migration, epidural fibrosis, animal, mouse, experiment |