Silk Fibroin Composite Decalcified Bone Matrix To Repair Bone Defect Of Animal Research

Objective:To evaluate and analysis bone repair situation, homemade diff er-rent proportion of silk fibroin composite decalcified bone matrix (DBM) in the bone defect model in ratMethod:1. The material preparation:① Preparation of silk fibroin:dissolved the home silk sericin protein off for solution and lyophilization; ② Preparation of DBM:The ulnar radial and femoral cortical bone of SD rats use deep cryogenic processing and made granular of bone;③ Preparation of Silk fibroin composite DBM material;The silk fibroin composite DBM in different concentration proportion of blending material to get experimental material (The content of DBM: 0%、10%、20%、40%、80%).2. Experimental method:A random sample of 72 only eight weeks of male SD rats divide into different concentration ratio of composite materials of silk fibroin and DBM.:Group A (do not contain DBM)、Group B (10%DBM)、Group C(20%DBM)、Group D(40%DBM). Group E (80%DBM), Group F (blank control), each group of 12;②Defect model making:,making 5 mm long bone defect model in rats with radial middle by jigsaw; ③ Material implanted:each materials about 5 mm wide 1.5 mm rectangle implanted into the defect model in rats;④ Test method:At 2,4,8 weeks after surgery, perform X-ray innormotopia; After radiography, remove the defect model to general investigation、HE dye、Masson dye、two kind of immunohistochemical (Osteocalcin, Collagen I) and grade adhesion degree, imaging and immunohistochemical score in the evaluation of bone defect healing, using Kruskal-Wallis H test to analyze the differences between groups of bone formation.Results:1. Application Hiteheock adhesion for the classification and statistical analysis after general observation:at the same time points adhesion between each score is no statistically significant difference (P>0.05), different time points within each group to compare the difference was statistically significant (P<0.05); 2. Application Lane -sandhu X-ray scores and statistical analysis: Each time point group D> E score is higher than group A, B, C, F score (P<0.01); 3. The two immunohistochemical (Osteocalci, Collagen I) were evaluated and done statistical analysis. After the experiment observation, Osteocalcin is positive reaction obviously in four weeks. Positive area of group D、E have obvious difference to other group (P<0.01).Group D and group E score no statistical significance (P>0.05).After the experiment observation, Type I collagen positive reaction obviously in four and eight weeks, positive area of group D and E have obvious difference with the other group. Group D is better than group E in four weeks(P<0.01)and both of them is no statistical difference in eight weeks (P>0.05)Conclusion:1. Silk fibroin composite decalcified bone matrix restorative materials has good osteogenetic capability.2. Silk fibroin composite decalcified bone matrix repair materials with the increase of the proportion of decalcified bone matrix, the osteogenesis performance will gradually increase in a certain extent.3. The ratio of 6:4 of silk fibroin and DBM (40% DBM) have better performance, to increase the proportion of decalcified bone matrix does not improve its performance.