基于PDGFR-β/PI3K/AKT信号通路探讨富血小板纤维蛋白修复大鼠牙槽骨缺损的作用机制

Explore the mechanism of platelet-rich fibrin in repairing rats alveolar bone defects based on PDGFR-β/PI3K/AKT signaling pathway

目的:基于血小板衍生生长因子受体-β/磷脂酰肌醇3激酶/蛋白激酶B(PDGFR-β/PI3K/AKT)信号通路探讨改良型/注射型富血小板纤维蛋白(A-PRF、i-PRF)联合骨替代材料(Bio-Oss)修复大鼠牙槽骨缺损的作用机制。方法:大鼠自体取血获得A-PRF、i-PRF,制备A-PRF/Bio-Oss、A-PRF/i-PRF/Bio-Oss复合物。大鼠分为Control组、A-PRF组、A-PRF/Bio-Oss组、A-PRF/i-PRF/Bio-Oss组,构建大鼠牙槽骨缺损模型,A-PRF组、A-PRF/Bio-Oss组、A-PRF/i-PRF/Bio-Oss组缺损处分别植入A-PRF、A-PRF/Bio-Oss、A-PRF/i-PRF/Bio-Oss复合物。于1个月后处死大鼠,并进行指标检测。Micro-CT、苏木精-伊红(HE)染色检测新骨形成情况;免疫组化染色、RT-qPCR检测牙槽骨组织成骨相关指标:Runt相关转录因子2(Runx2)、骨形态发生蛋白-2(BMP-2)、骨钙素(OCN)、I型胶原(Col I)和成血管相关指标:血小板-内皮细胞粘附分子(CD31)、血管内皮生长因子A(VEGFA)mRNA和蛋白表达;Western blot检测牙槽骨组织PDGFR-β/PI3K/AKT信号通路相关蛋白表达。结果:A-PRF/i-PRF/Bio-Oss组牙槽骨新生骨量、新生血管、Runx2、BMP-2、OCN、Col I、CD31、VEGFA、PDGFR-β、p-PI3K/PI3K、p-AKT/AKT表达量明显高于A-PRF/Bio-Oss组、A-PRF组、Control组(均P<0.05)。结论:A-PRF/i-PRF/Bio-Oss复合物通过促进骨形成和血管形成,促进大鼠牙槽骨缺损的再生修复,可能与激活PDGFR-β/PI3K/AKT信号通路有关。

Objective:To explore the mechanism of advanced/injectable platelet rich fibrin(A-PRF,i-PRF)combined with bone substitute material(Bio-Oss)in repairing rats alveolar bone defect based on platelet-derived growth factor receptor-β/phosphatidylinositol 3-kinase/protein kinase B(PDGFR-β/PI3K/AKT)signaling pathway.Methods:Rats autologous blood were collected to obtain A-PRF and i-PRF,and A-PRF/Bio-Oss,A-PRF/i-PRF/Bio-Oss complexes were prepared.Rats were divided into Control group,A-PRF group,A-PRF/Bio-Oss group and A-PRF/i-PRF/Bio-Oss group,and rats were constructed alveolar bone defect model.A-PRF group,A-PRF/Bio-Oss group and A-PRF/i-PRF/Bio-Oss group were implanted respectively with A-PRF,A-PRF/Bio-Oss and A-PRF/i-PRF/Bio-Oss complexes at the defect site.After 1 month,the rats were euthanized and indicators were tested.Micro-CT and HE staining were used to detect the new bone formation.Immunohistochemical staining and RT-qPCR were used to detect the expression of osteogenesis related indicators of alveolar bone tissue:Runt related transcription factor 2(Runx2),bone morphogenesis protein-2(BMP-2),osteocalcin(OCN),collagen I(Col I)and angiogenesis related indicators:platelet endothelial cell adhesion molecule-1(CD31),Vascular endothelial growth factor A(VEGFA)mRNA and protein.Western blot was used to detect the expression of PDGFR-β/PI3K/AKT signaling pathway related proteins in alveolar bone tissue.Results:New bone formation,new angiogenesis,the expression of Runx2,BMP-2,OCN,Col I,CD31,VEGFA,PDGFR-β,p-PI3K/PI3K,p-AKT/AKT levels in alveolar bone of A-PRF/i-PRF/Bio-Oss group were significantly higher than those of A-PRF/Bio-Oss group,A-PRF group and Control group(all P<0.05).Conclusion:A-PRF/i-PRF/Bio-Oss complex promoted the regeneration and repair of alveolar bone defects in rats via promoting bone formation and angiogenesis,which may be related to the activation of PDGFR-β/PI3K/AKT signaling pathway.