PRF复合组织工程骨修复牙周骨缺损中基因的表达及意义

Gene expression and clinical significance of PRF composite tissue-engineered bone in the repair of periodontal bone defects

目的探讨组织工程煅烧骨复合富血小板纤维蛋白(PRF)修复牙周骨缺损中BMP-2、骨保护素(OPG)和核因子B受体活化因子配体(RANKL)的表达及意义。方法将36只新西兰大白兔随机均分为4组,制备单侧牙周骨缺损模型,其中3组于骨缺损处分别植入煅烧骨、Bio-oss骨和煅烧骨/PRF复合物,以未植入任何材料者作为空白对照。术后4,8,12周处死动物,获取完整标本,进行大体、Masson染色及免疫组化观察。结果 Masson染色:煅烧骨/PRF组术后8周时即见部分鲜红色成熟骨形成,骨成熟速度明显优于其他3组。免疫组化观察:术后4周时煅烧骨/PRF组BMP-2、OPG、RANKL强阳性表达,数目高于其他3组,差异具有统计学意义(P<0.05)。RANKL阳性细胞的表达在骨改建过程中有不同程度的降低,煅烧骨/PRF组中组中RANKL下降最为明显,差异有统计学意义(P<0.05)。结论煅烧骨/PRF组修复牙周骨缺损会增加BMP-2、OPG、RANKL的表达,通过促进成骨细胞分化成熟而刺激OPG表达升高,抑制破骨细胞,有助于加快牙周骨改建,缩短骨愈合过程。

Objective To observe expression and significance of bone morphogenetic protein-2（ BMP-2）,osteoprotegerin（ OPG） and receptor activator of NF kappa B ligand（ RANKL） protein in periodontal bone defects repair with calcined bone compounded with platelet-rich fibrin（ PRF） in rabbits. Methods A total of 36 healthy rabbits were randomly divided into four groups,and unilateral periodontal tissue defect model was established in each group. These four groups were respectively treated with calcined bone,Bio-oss bone and calcined bone/PRF,and nothing（ blank control group）. After 4,8 and 12 weeks,animals were sacrificed respectively for a complete specimen used in gross,Masson staining and Immunohistochemical observation. Results Masson staining： In calcined bone/PRF group after 8 weeks,visible partial bright red mature bone formed,and bone maturation rate was better than the other three groups. Immunohistochemical observation： After 4 weeks in calcined bone/PRF group,BMP-2,OPG,RANKL were strongly positive,and the number was higher than the other three groups with statistically significant differences（ P0. 05）. Expression of RANKL-positive cells declined in bone remodeling process in varying degrees,while in calcined bone/PRF group,RANKL declined most obviously with statistically significant differences（ P0. 05）. Conclusion Repair of periodontal bone defects of calcined bone/PRF group will increase the expression of BMP-2,OPG,RANKL. Through the promotion of osteoblast differentiation,OPG expression can be stimulated,osteoclast can be inhibited,which help accelerate the periodontal bone remodeling and shorten the healing process.