| Objective:(1)To establish an animal model of implants restoration in anterior New Zealand rabbits,and to construct tissue engineered implants using tissue engineering to combine TGF-β3 with DPSCs with implants.(2)By comparing the combined effects of TGF-β3 and DPSCs during the repair of anterior teeth of New Zealand rabbits in each group,the mechanism of TGF-β3 in promoting bone healing was investigated.Methods:(1)Select the removal of New Zealand rabbit under the anterior teeth of the lower anterior teeth prosthesis planting model;(2)experimental group for the implant and bone interstitial filled with TGF-β3,DPSCs,bio-oss bone meal mixture,the control group for the implant and the bone interstitial filled with DPSCs,bio-oss bone meal mixture,the blank group for the implant and interstitial filled with phosphate buffer,bio-oss bone powder mixture;(3)set 2 weeks,4 weeks,8 weeks,12 weeks,4 time limit,each limit of6 animals.Animals were sacrificed at each point and the implant repair area were generally observed.The free mandible and peri-implant tissue were observed histologically.Immunohistochemistry and real-time fluorescence quantitative PCR were used to detect the expression of TGF-β3 in the implants Promote the mechanism of bone tissue healing.Results:(1)histological observation at 2 weeks,the experimental group compared with control group,blank group appeared more osteoblasts,bone lacuna formation and neovascularization area,new bone forming new bone area,the formation of regional boundaries more clearly;at 4 weeks,the experimental group compared with control group,blank group appeared a lot of new bone very low density formation,vascular connective tissue with new bone cells in areas with honeycomb,a decrease in the number of osteoblasts,bone cell number increased significantly,with early bone tissuerepair;at 8 weeks,the experimental group compared with control group,blank group,a large number of osteoblasts were fixed into bone cells in bone matrix in the formation of woven bone trabecular immature,with the increase of bone matrix minerals and organic matter in the bone matrix,mature lamellar bone obvious calcification,so that the woven bone was replaced complete osseointegration Repair.At 12 weeks,the experimental group had basically formed lamellar bone.There was no obvious boundary between the new bone and the old bone.The number of bone cells,the number of bone cells and the vascularization of the control group were not as good as those of the experimental group,and the time of osteogenesis was slower than that of the experimental group for 2-4 weeks.And the experimental group and the control group have more bone area than the blank group,and the healing condition is better.(2)immunohistochemical staining: the osteoblasts around the implant in the experimental group were more and more early than the other two groups,regardless of the number of cells.The experimental group,control group and blank group is significantly difference(P < 0.05),so that the osteogenic effect was better than the control group,blank group,shows the TGF-beta combined with DPSC under the effect of implant osseointegration was completed ahead of schedule,(3)real time fluorescent quantitative PCR found in bone cycle three within months,the experimental group of RUNX2 gene was significantly higher than the control group(P <0.05),proved that TGF-beta 3 effectively stimulate the body to promote osteoblast proliferation,high expression of bone RUNX2 gene,the expression of COL-I in the experimental group than the control group,indicating the osteoblasts in the experimental group has formed osteoblasts and polarization,while the control group group and blank group had no significant difference,may be due to concentration time is not to stimulate cell differentiation and expression of COL-I.Conclusion:(1)it is feasible to construct a tissue engineered implant with TGF-β3 combined with the implant.(2)in vivo experiments in animals showed that exogenous TGF-β3combined with a mixture of DPSCs and Bio-oss bone powder could promote and accelerate bone tissue formation.(3)this technique will provide new ideas and methods to solve the problem of long bone healing time and high failure rate for the patients with bone metabolic diseases. |
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