RESUMO
BACKGROUND: Preliminary study has found that osteopractic total flavone can promote osteogenic differentiation of MC3T3-E1 cells on the surface of nano-bone material, but the underlying mechanism needs to be studied in depth. OBJECTIVE: To investigate the actin mechanism of osteopractic total flavone combined with nano-bone material on MC3T3-E1 cells. METHODS: MC3T3-E1 cells were co-cultured with nano-bone material, and 100 mg/L and 250 mg/L osteopractic total flavone were treated as drug intervention, including 10 μg/L transforming growth factor-β as positive control. The samples were divided into eight groups: (1) Normal group; (2) DKK1 group: Wnt pathway inhibitor DKK1 (0.1 mg/L) blocks Wnt/β-catenin signaling pathway; (3) DKK1+transforming growth factor-β group; (4) DKK1+100 mg/L osteopractic total flavone group; (5) DKK1+250 mg/L osteopractic total flavone group; (6) DKK1+ nano-hydroxyapatite/collagen+transforming growth factor-β group; (7) DKK1+nano-hydroxyapatite/collagen+100 mg/L osteopractic total flavone group; (8) DKK1+nano-hydroxyapatite/collagen+250 mg/L osteopractic total flavone group. Cells in each group were harvested after 24 and 48 hours of intervention. Immunofluorescence labeling was used to observe the binding of Wnt and LRP in osteoblasts in the Wnt/β-catenin pathway. The expression of β-catenin, LRP 5, GSK-3β, Cyclin D1, and RUNX2 was detected by real-time polymerase chain reaction and western blot assay. RESULTS AND CONCLUSION: (1) Confocal laser scanning microscope showed that obvious brown and yellow staining was shown in the DKK1+transforming growth factor-β group, DKK1+250 mg/L osteopractic total flavone group, DKK1+nano-hydroxyapatite/ collagen+transforming growth factor-β group, and DKK1+nano-hydroxyapatite/collagen+250 mg/L osteopractic total flavone group, indicating that Wnt and LRP combined better than other groups. (2) Real-time polymerase chain reaction and western blot assay results showed that osteopractic total flavone could promote the expression of β-catenin, LRP5 and RUNX2, and downregulated GSK3β expression. These findings confirm that osteopractic total flavone can promote the differentiation and proliferation of osteoblasts by activating the Wnt/β-catenin signaling pathway. Gene activation induced by osteopractic total flavone was dose-dependent.
RESUMO
Objective To explore the application value of single-door laminoplasty via Y type nano-bone plate in treating multilevel cervical spondylotic myelopathy (MCSM) and analyze the short-term efficacy.Methods From January 2013 to December 2016,79 cases of MCSM were treated with single-door laminoplasty via Y type nano-bone plate to evaluate the improvement of post-operative neurological function by the Japanese Orthopaedic Association (JOA) evaluation system.We also measured cervical curvature of cervical X-ray and C5 sagittal diameter of the spinal canal before operation and 6 months after operation to understand the maintenance and enlargement of the spinal canal.The improvement degree of spinal cord compression was evaluated by preoperative and postoperative cervical MRI.Osseous healing on the open door side and the door shaft side was observed with the aid of three-dimensional spiral CT.Results Follow-up ranged from 6 to 36 months,with an average of (20.4±7.9)months.Preoperative JOA score was (8.6 ± 1.3) points and JOA score 6 months after operation was (14.3 ± 1.5)points (P ＜ 0.05).JOA improvement rate was (68.6 ± 15.8)％;postoperative follow-up X-ray and threedimensional spiral CT showed that the spinal canal had satisfying enlargement,the door shaft side all had osseous healing,the open door side osseous healing was not obvious,and there was no lamina collapse or reclosing.Sagittal diameter of theC spinal canal was (8.9±l.1)mm before operation and (15.1±l.1)mm 6 months after operation (P＜0.05).The spinal canal enlargement rate was (70.8±22.3)％,cervical curvature was (14.8± 7.0)°preoperatively and (15.1±6.7)°postoperatively with no significant difference (P＞0.05).Conclusion EOLP via Y type nano-bone plate is safe and efficacious in treating MCSM.It not only provides a good immediate fixation,but also provides the possibility for the open door side lateral osseous fusion.