Preparation And Experimental Study Of Silk Fibroin/Human Bone Morphogenetic Protein 2 Active Peptide

Part I Preparation and osteogenic property analysis of silk fibroin/human bone morphogenetic protein 2 active peptideObjcective:Prepare silk fibroin/human bone morphogenetic protein 2 active peptide(SF/BMP2 active peptide).Study their effects on cell osteogenic activity and provide material basis for subsequent trials.Methods:Silk fibroin/human bone morphogenetic protein 2 was successfully prepared by solid-phase peptide synthesis.With gelatin sponge as the scaffold material, composite SF/BMP2 active peptide between leachate and bone marrow mesenchymal stem cells (BMSCs) training. Using CCK-8 measured the status of cell growth and toxicity.The osteogenesis ability of artificial bone were observed by alizarin red staining and ALP by training with BMSCs for 2 weeks.Result:Using solid-phase synthesis become successful preparation of silk fibroin/BMP2 active peptide, SF/BMP2 active peptide extract on the growth of cell proliferation and normal training group have no statistical difference(P>0.05).The alizarin red and ALP stain dyeing showed that bone marrow mesenchymal stem cells after active peptides induced the expression between osteoblasts characteristics.Concluson:SF/BMP2 active peptide can induce the cells that adhesion and growth on the surface of it to osteogenic differentiation. The experimental result show that SF/BMP2 active peptide can be used as a bone tissue active factor.Part II:Ectopic osteoinduction study of silk fibroin/human bone morphogenetic protein 2 active peptideObjective:To and evaluate the ectopic bone formation performance.Methods:The concentration of SF/BMP2 active peptide was measured by BCA at different time points in the phosphate buffer,calculate the cumulative release of SF/BMP2 active peptide.The sponge group particles were implanted in 24 rats thigh muscle tissue and the Gelatin sponge group was the control group. The rats were killed during 6 weeks and the thigh muscle tissues were observed after CT and HE staining.Result:SF/BMP2 active peptide release meets biphasic kinetics of drug release pattern. Tissue morphology and CT found that lots of cartilage cells, chondrocytes and bone formation were generated in experimental group.Conclusion:The regenerated silk fibroin/human bone morphogenetic protein 2 active peptide which carried the BMP-2 can not only maintain the activity of BMP-2 but also slow its release and can continue to induce bone formation.