Abstract
Objective To prepare silk fibroin (SF) microspheres loaded with recombinant human bone morphogenetic protein-2 (rhBMP-2) and evaluate the ectopic bone formation performance.Methods The rhBMP-2-loaded fibroin particles were prepared with freeze-drying methods.The microspheres were observed by scanning electron microscope.They were prepared by the volume ratio of silk fibroin and cosslinking agent was 1∶20,5∶20,10∶20,and the mass ratio of rhBMP-2 and SF was 0.5 μg∶1 mg.The concentration of rhBMP-2 was measured by enzyme linked immunosorbent assay (ELISA) at different time points in the phosphate buffer (PBS),calculate the cumulative release of rhBMP-2 and microsphere drug loading rate.The SF microspheres and SF/rhBMP-2 particles were implanted in 24 rats thigh muscle tissue and the SF microspheres group was the control group.The rats were killed during 4 weeks and the thigh muscle tissue were observated after hematoxylin and eosin (HE) staining.Results The shape of SF/rhBMP-2 microspheres were regular circular and the diameter decreases with the concentration of ethanol increase.Their diameters were (835.0 ±97.23),(715.30 ± 193.80),(398.70 ±99.86) nm.The rhBMP-2 release meets biphasic kinetics of drug release pattern.Drug loading were (4.54 ±0.13)％,(4.55 ±0.05)％,(4.53 ± 0.08) ％ with no significant difference (P ＞ 0.05).Tissue morphology found that lots of cartilage cells and chondrocytes were generated in experimental group.Conclusion The regenerated silk fibroin microspheres which carried the rhBMP-2 can not only maintain the activity of rhBMP-2 but also slow its release and can continue to induce bone formation. Key words: Silk fibroin; Microspheres; Bone morphogenetic protein-2; Ectopic bone formation
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