共表达骨保护素和碱性成纤维细胞生长因子的重组腺病毒对牙槽骨缺损修复的实验研究

目的以含有骨保护素(OPG)和碱性成纤维细胞生长因子(bFGF)的共表达重组腺病毒感染牙龈及牙周膜成纤维细胞,观察OPG与bFGF表达的情况;以重组腺病毒结合胶原膜置入犬牙槽骨缺损处,观察其对牙槽骨缺损修复的影响。方法培养牙龈及牙周膜成纤维细胞,感染重组腺病毒,运用Western blot、RT-PCR分别检测2种细胞中OPG及bFGF的表达,并以正常培养的牙龈及牙周膜成纤维细胞作为对照组。制造犬下颌前磨牙根分叉处牙槽骨缺损模型,右侧作为空白对照组(n=2)和材料对照组(n=4),左侧作为实验组(n=6),光镜观察骨缺损处牙槽骨新生情况,免疫组织化学方法检测OPG和bFGF在犬牙周组织中的表达。结果细胞培养结果显示,感染重组腺病毒的牙龈和牙周膜成纤维细胞,OPG及bFGF的表达在蛋白和基因水平上均明显高于正常对照组;动物实验结果显示,实验组骨缺损处有多量新生牙槽骨生长,其OPG和bFGF较材料对照组和空白对照组均有较强的表达。结论 OPG和bFGF共表达的重组腺病毒可提高牙龈及牙周膜成纤维细胞OPG和bFGF的表达,对牙槽骨缺损修复起促进作用。

Experimental study of recombinant adenovirus co-expressing osteoprotegerin and basic fibroblast growth factor on the repair of alveolar bone defects

Objective To explore the expression of osteoprotegerin(OPG) and basic fibroblast growth factor(bFGF) after the infection of recombinant adenovirus encoding OPG and bFGF in the gingival fibroblasts(GFs) and periodontal ligament fibroblasts(PDLCs);And to observe the repairing effect on the alveolar bone defects of recombinant adenovirus with collagen membrane.Methods GFs and PDLCs were cultured and infected by the recombinant adenovirus,in which the expression of OPG and bFGF were detected by Western blot and RT-PCR,simultaneously cultured normal GFs and PDLCs served as the control groups;Alveolar bone defects model of root furcation area of canine mandibular premolar were prepared,the right side was selected as the blank control group(n = 2) and material control group(n = 4),which the left as the experimental group(n = 6).The light microscopic was used to observe the newborn alveolar bone in bone defect area,and the expression of OPG and bFGF in canine periodontal tissues was detected by immunohistochemistry.Results The expression of OPG and bFGF on the level of gene and protein in infected cells were significantly higher than those of the normal control groups.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 that of the material control group and blank control group.Conclusion The recombinant adenovirus co-expressing OPG and bFGF may improve the expression of OPG and bFGF in GFs and PDLCs,which can promote the repair of alveolar bone defects.