The Experimental Study Of Recombinant Adenovirus Encoding Osteoprotegerin And Basic Fibroblast Growth Factor On The Repair Of Alveolar Bone Defects

Periodontal disease is a chronic infectious diseases occur in the periodontal support, clinically, there is no effective clinical treatment at present. With the rise and development of tissue engineering and gene therapy technology, pathogenic factors in the treatment of periodontal disease can be removed through surgery, then use of gene therapy to express cytokines to promote tissue repair and regeneration. The study found that in the field of gene therapy, at the same time transfer and expression of multiple exogenous genes value higher than a single gene.This project from bone resorption-the redevelopment and regeneration mechanism starting,to explore effective methods of treatment of periodontal disease:Through amplification osteoprotegerin(OPG) to inhibition osteoclast function and basic fibroblast growth factor(bFGF) to promote tissue hyperplasia, constructed recombinant adenovirus containing the OPG and bFGF gene. Use of co-expression of the recombinant adenovirus technology to increase the secretion of OPG in the periodontal tissue lesions, thereby inhibiting RANKL function, blocking the proliferation, differentiation and maturation of osteoclasts; Simultaneously application of bFGF to promote cell proliferation, to treatment of severe periodontal disease periodontal tissue defects.Objective To explore the infection of recombinant adenovirus encoding OPG and bFGF in the gingival fibroblasts(GFs) and periodontal ligament fibroblasts(PDLCs), observed the expression of OPG and bFGF; and combined recombinant adenovirus with collagen membrane, which was placed into the alveolar bone defects of canine to observe the repairing effect on the alveolar bone defects.MethodsGFs and PDLCs were cultured and infected by the recombinant adenovirus, Western Blot and RT-PCR were used to detect the expression of OPG and bFGF in these two kinds of cells, simultaneously normal cultured GFs and PDLCs as the control groups; Alveolar bone defects model of root furcation area of canine mandibular premolar were prepared, take left as the experimental group, right as the blank control group and material control group. Experimental group(n=6):dropwise recombinant adenovirus collagen membrane placed in bone defects, suture gingival flap; Material control group(n=4):not dropwise recombinant adenovirus collagen membrane placed in bone defects, suture gingival flap; Blank control group(n=2):direct suture gingival flap. The light microscopic was used to observe the newborn of alveolar bone in bone defect area, and the expression of OPG and bFGF in canine periodontal tissues was detected by immunohistochemistry.ResultsThe expression of OPG and bFGF on the level of gene and protein in infected cells were significantly higher than in the normal control groups which was showed by the cell culture result. Animal experiments showed that bone defect had large amounts of new bone growth in the experimental group, OPG and bFGF in the experimental group had a stronger expression than in the material control group nor blank control group. ConclusionThe recombinant adenovirus encoding OPG and bFGF may improve the expression of OPG and bFGF in GFs and PDLCs, and can promote the repair of alveolar bone defects.