摘要
目的:基于c-Jun氨基末端蛋白激酶(JNK)通路研究改良型富血小板纤维蛋白(A-PRF)与β-三磷酸钙(β-TCP)复合物对兔股骨缺损区域的诱导成骨效果,为临床治疗骨缺损提供实验基础。方法:选取24只雄性新西兰大白兔构建股骨缺损模型,动物分为模型组、1:1复合物组(A-PRF:β-TCP=1:1)、2:1复合物组(A-PRF:β-TCP=2:1)与4:1复合物组(A-PRF:β-TCP=4:1),每组各6只。模型组对骨缺损区域不作填充处理,复合物组填充对应的材料,8周后处死动物收集指标。肉眼观察骨缺损区域的修复情况,经苏木精-伊红(HE)染色观察新生骨组织修复情况,以酶联免疫吸附法(Elisa)检测血清中碱性磷酸酶(ALP)和骨钙素(OCN)的含量,以蛋白质免疫印迹法(WesternBlot)检测骨组织中骨形成蛋白9(BMP-9)、JNK与p-JNK蛋白的表达情况。结果:与模型组相比,复合物组填充后骨缺损区域新生血管增多,成骨效果明显,炎症反应少,血清中ALP与OCN含量显著升高,骨组织中BMP-9与p-JNK蛋白表达水平显著升高(P<0.05)。结论:A-PRF与β-TCP复合物能通过激活JNK通路、诱导BNP-9蛋白表达,促进骨缺损后诱导成骨,当复合物比例为2:1时效果最佳。
Objective:To study the osteogenic effect of modified platelet-rich fibrin(A-PRF)andβ-tricalcium phosphate(β-TCP)complex on rabbit femoral defect based on c-Jun amino terminal protein kinase(JNK)pathway,and to provide experimental basis for clinical treatment of bone defect.Methods:Twenty-four male New Zealand white rabbits were selected to construct the femur defect model.The animals were divided into model group,1:1 complex group(A-PRF:β-TCP=1:1),2:1 complex group(A-PRF:β-TCP=2:1)and 4:1 complex group(A-PRF:β-TCP=4:1),with 6 rabbits in each group.The bone defect area was not filled in the model group,and the composite group was filled with the corresponding material.8 weeks later,the animals were killed to collect indicators.The repair of the bone defect area was observed by naked eye,the repair of the new bone tissue was observed by hematoxylin-eosin(HE)staining,and the contents of alkaline phosphatase(ALP)and osteocalcin(OCN)in serum were detected by enzym-linked immunosorbent assay(Elisa).The expressions of bone morphogenetic protein 9(BMP-9),JNK and p-JNK in bone tissue were detected by Western blotting(WesternBlot).Results:Compared with the model group,the complex group was filled with more neovascularization in the bone defect area,the osteogenesis effect was obvious,the inflammatory reaction was less,the serum ALP and OCN contents were significantly increased,and the protein expression levels of BMP-9 and P-JNK in bone tissue were significantly increased(P<0.05).Conclusion:A-PRF andβ-TCP complex can promote osteogenesis after bone defect by activating JNK pathway and inducing BNP-9 protein expression,and the best effect is achieved when the ratio of c omplex is 2:1.
作者
付冬梅
王浪
周婧
王劲
杨昕
李素兰
兰红
FU Dongmei;WANG Lang;ZHOU Jing;WANG Jin;YANG Xin;LI Sulan;LAN Hong(Department of Stomatology,Ziyang Yanjiang District People's Hospital,Ziyang 641300)
基金
富血小板纤维蛋白联合骨替代材料对成骨细胞生物学行为及成骨影响的体外对比研究,项目编号:Zykjjsc20-yyjc-2020-09
改良型富血小板纤维蛋白与β三磷酸钙混合物在骨缺损中的研究,项目编号:17ZD027。
