改良型富血小板纤维蛋白与β⁃磷酸三钙复合物诱导成骨作用与机制  

Study on the role and mechanism of osteogenesis induced by advanced platelet⁃rich fibrin andβ⁃tricalcium phosphate complex

作　　者：付冬梅[1] 周婧 王浪 杨昕 兰红 李素兰 王劲 方婕 FU Dongmei;ZHOU Jing;WANG Lang;YANG Xin;LAN Hong;LI Sulan;WANG Jin;FANG Jie(Department of Stomatology,Yanjiang District People's Hospital,Ziyang City,Ziyang 641300,China;Department of Orthodontics,West China Hospital of Stomatology,Sichuan University,Chengdu 610041,China)

机构地区：[1]资阳市雁江区人民医院口腔科,四川资阳641300 [2]四川大学华西口腔医院正畸科,四川成都610041

出　　处：《口腔疾病防治》2023年第4期237-244,共8页Journal of Prevention and Treatment for Stomatological Diseases

基　　金：四川省科学技术厅项目(23ZDYF2009);四川省卫生和计划生育委员会科研课题(17ZD027);资阳市科技计划项目(Zykjjsc20⁃yyjc⁃2020⁃09)。

摘　　要：目的探讨改良型富血小板纤维蛋白(advanced platelet⁃rich fibrin,A⁃PRF)与β⁃磷酸三钙(β⁃tricalcium phosphate,β⁃TCP)构成比例对兔股骨缺损模型骨形成的作用及机制,为临床治疗骨缺损提供新思路。方法24只新西兰大白兔构建骨缺损模型并分为模型组、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6只。复合物组在骨缺损处填充复合材料,模型组不充填材料,8周后安乐法处死动物采标本。采用micro⁃CT测定股骨缺损区新生骨的骨密度(bone mineral densi⁃ty,BMD)、骨体积分数(bone volume fraction,BV/TV)、骨小梁厚度(trabecular thickness,Tb.Th)、骨小梁分离度(trabecular separation/spacing,Tb.Sp)。HE染色观察新骨及新生血管形成情况;扫描电镜观察新生骨组织的形态变化;酶联免疫法检测新生股骨组织中磷酸化丝裂原活化蛋白激酶p38(phospho⁃p38 mitogen activated pro⁃tein kinase,p⁃p38MAPK)、CCAAT/增强子结合蛋白同源蛋白(CCAAT/enhancer⁃binding protein homologous pro⁃tein,CHOP)、磷酸化含半胱氨酸的天冬氨酸蛋白水解酶3(phospho⁃cysteine aspartic protease⁃3,p⁃Caspase3)含量;实时定量PCR检测新生骨组织中骨保护素(osteoprotegerin,OPG)、骨形态发生蛋白⁃2(bone morphogenetic protein⁃2,BMP⁃2)、核因子κ⁃B配体受体致活剂(receptor activator of nuclear factor kappa⁃B ligand,RANKL)、p38MAPK的mRNA表达量;免疫组织化学法观察新生骨组织中OPG、BMP⁃2、RANKL、p⁃p38MAPK、p⁃Caspase3蛋白表达情况;荧光Tunel染色观察新生股骨组织中细胞凋亡情况。结果模型组股骨缺损区域骨形成缓慢,成骨质量差。与模型组相比,复合物组动物股骨缺损区域成骨情况良好,BMD、BV.TV、Tb.Th、Tb.N升高,Tb.Sp下降,新生股骨组织中见新生毛细血管形成;p⁃p38MAPK、CHOP、p⁃Caspase3蛋白表达降低,OPG、BMP⁃2的mRNA与蛋白表Objective To investigate the role and mechanism of bone formation caused by the ratio of advanced platelet⁃rich fibrin(A⁃PRF)andβ⁃tricalcium phosphate(β⁃TCP)in rabbit femur defect model,which provides a new idea for clinical treatment of bone defect.Methods Twenty⁃four New Zealand white rabbits 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.Femoral defect models were constructed in each group.In the com⁃posite group,the bone defect was filled with composite material,while in the model group,no material was filled.After 8 weeks,the animals were euthanized and specimens were collected.Bone mineral density(BMD),bone volume fraction(BV/TV),trabecular thickness(Tb.Th),trabecular separation(Tb.SP)and trabecular number(Tb.N)in femoral defect tis⁃sue were measured by micro⁃CT and photographed.Hematoxylin⁃eosin staining was used to detect the pathological changes of new bone tissue.The morphological changes of the new bone tissue were observed by scanning electron mi⁃croscopy.Determination of phospho⁃mitogen activated protein kinase p38(p⁃p38MAPK),CCAAT/enhancer binding pro⁃tein homologous protein(CHOP)and phospho⁃cysteine aspartic protease⁃3(p⁃Caspase3)in newborn femur by ELISA.The mRNA expressions of osteoprotegerin(OPG),bone morphogenetic protein⁃2(BMP⁃2),receptor activator of nuclear factor kappa⁃B ligand(RANKL)and p38MAPK were detected by real⁃time quantitative PCR.The expression of OPG,BMP⁃2,RANKL,p⁃p38MAPK and p⁃Caspase3 protein in the new bone tissue was observed by immunohistochemistry.Results In the model group,bone formation in the femoral defect area was slow and osteogenic quality was poor.Com⁃pared with the model group,the bone formation and neocapillaries of femoral defect area in the complex group was good,BMD,BV.TV,Tb.Th,Tb.N were increased,and Tb.Sp were decreased,the expressions of p⁃p38MAPK,CHOP and p

关 键 词：改良型富血小板纤维蛋白 β⁃磷酸三钙 骨缺损 成骨分化 丝裂原活化蛋白激酶P38 CCAAT/增强子结合蛋白同源蛋白 含半胱氨酸的天冬氨酸蛋白水解酶3 骨保护素 骨形态发生蛋白⁃2 核因子κ⁃B配体受体致活剂 

分 类 号：R78[医药卫生—口腔医学]