| Objective: To investigate the ectopic osteogenic activity of KLD-12 polypeptide / recombinant human bone morphogenetic protein-2(rh BMP-2)nanofiber gel combine with bone barrow besenchymal stem cel s(BMSCs)in the gluteal muscle of rabbits.Methods: Group: 48 New Zealand white rabbits,6 moons old,weight 3.0-3.5kg,male and female,were randomly divided into A,B,C,D group,each group of 12.Group A was KLD-12 polypeptide / rh BMP-2/BMSCs group;group B was KLD-12 polypeptide / rh BMP-2 group;group C was rh BMP-2 group,group D was KLD-12 group.Implant the compound into the gluteal muscles of each group.The ectopic bone was tested by general observation,X-ray,Micro-CT,histomorphology and alkaline phosphatase activity(ALP)at 4 weeks,8 weeks and 12 weeks after operation,Results:(1)x ray examination: osteogenesis images was seen in group A,B,C,the formation of bone gradually increased with time;group D: no osteogenesis image.(2)Micro-CT: the bone volume fraction,the number of trabecular bone,the tissue mineral density and the thickness of trabecular bone gradually increased with time.12 weeks,the bone volume fraction BV/TV(%):group A 4.274±0.134?group B3.841±0.131?group C 3.689±0.321;the number of trabecular bone TB.N(?/mm3):group A 0.712±0.047?group B 0.616±0.032?group C 0.569±0.045;tissue mineral density TMD(mg/cm3):group A 0.084±0.011?group B 0.097±0.012?group C 0.102±0.041;the thickness of trabecular bone Tb.Th(mm):group A 0.104±0.022,group B 0.092±0.013,group C 0.083±0.023,the differences are statistically significant(P<0.05).(3)general observation: At 4 weeks,the hardness of the bone was low in group A,B and C,and at 8 weeks and 12 weeks,the hardness of bone in group A,B and C was higher than before.Group D: no bone found.(4)HE staining: 100×,at 4 weeks: group A showed more amount of woven bone,group B showed a small amount o f trabecular bone formation,group C showed a small amount of woven bone;8 weeks: group A showed more mature lamellar bone and woven bone,group B showed a large number of woven bone,group C showed a few mature lamellar bone and a large number of woven bone;12 weeks: group A and group B showed mature bone tissue,group C showed mature bone tissue and a small amount of woven bone.(5)Giemsa staining: 100×,4 weeks: group A showed a few deep blue mature bone,the group B showed more amount of lake blue bo ne,group C showed light blue new mineralized bone at 8 weeks;group A showed more deep blue mature bone,the group B showed a few deep blue mature bone,group C showed a few deep blue mature bone,and more amount of light blue new mineralized bone.at 12 weeks: group A showed a large number of deep blue mature bone tissue,group B showed more amount of deep blue mature bone,group C showed deep blue bone and light blue new mineralized bone.(6)ALP activity(king unit/gprot): at 4 weeks:group A 14.466±0.136?group B 13.136±0.292?group C 13.964±0.293;at 8 weeks:group A 16.281±0.233?group B 15.771±0.155?group C 14.565±0.383;at 12 weeks:group A 20.236±0.351?group B17.697±0.533?group C 16.981±0.246,the differences are statistically significant(P<0.05).Conclusion: the ectopic osteogenic activity of KLD-12/rh BMP-2 nanofiber gel combine with BMSCs in the gluteal muscle of rabbits was good. |
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