| Objective: Prepared silk fibroin/BMP-2 active polypeptide(SF/BMP-2 active polypeptide), to research the drug release curve and the role in promoting MC3T3-E1 proliferation and differentiation, and lumbar fusion in rats.Methods:(1) in vitro: SF/BMP-2 active polypeptide was prepared by solid-phase synthesis method, gelatin sponge was used as the scaffold material, SF/BMP-2 active polypeptide/gelatin sponge composite material was constructed. The release curve of SF/BMP-2 active peptide at different time points was measured by BCA method. MC3T3-E1 cell cultivated by the composite material extract solution, observed the proliferation of cell by CCK-8 method, The osteogenesis differentiation ability was observed by alizarin red staining and alkaline phosphatase(ALP) staining.(2) in vivo: We divided SD rats into three groups: the control group(G1), the BMP-2 loaded gelatin sponge group(G2),and SF/BMP-2 active polypeptide/gelatin sponge group(G3). 1 month or 3 months after surgery, the spines fusion of the rats were evaluated by manual palpation, micro-computed tomography(micro-CT), haematoxylin and eosin(H&E) staining.Results:(1)In vitro: The release curve of SF/BMP-2 active polypeptide/gelatin sponge composite accorded with the sustained release law. MC3T3-E1 can grow and proliferation in SF/BMP-2 active polypeptide extract solution. The alizarin red and ALP staining showed that MC3T3-E1 cell expressed the osteogenic ability.(2) In vivo: Manual palpation showed that the SF/BMP-2 active polypeptide/gelatin sponge group had obvious lumbar posterolateral fusion. CT detection found new bone formation in posterolateral lumbar. Histomorphology observed new bone trabecula formation.Conclusion: SF/BMP-2 active polypeptide/gelatin sponge composite material had certain sustained release effect. It can promote the proliferation and osteogenetic differentiation of MC3T3-E1 cell. It had a certain effect on enhancing SD rat lumbar posterolateral fusion. |
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