The Effect and Mechanism of Collagen Membranes Loaded With Fibrinogen on the Healing of Femur Fractures in Rats

Yang Ping; Lei Zhijie; Yu Jianglin; Qiu Xusheng; Chen Yixin; Qi Xiaoyang

Abstract：Objective To investigate the effect and mechanism of the collagen membrane loaded with fibrinogen on healing of femoral fractures in rats.Methods The rat femur fracture models were randomly divided into three groups:control group,collagen membrane group,and fibrinogen-loaded collagen membrane group.At 8 weeks post-surgery,the affected femurs of the rats were analyzed using Micro-CT scanning to assess callus formation.Primary bone marrow mesenchymal stem cells(BMSC)were divided into osteogenic control group,fibrinogen+osteogenic induction group,fibrinogen+PNU-74654+osteogenic induction group.BCIP/NBT staining was used to detect the level of alkaline phosphatases(ALP)in BMSC.Wsetern blot was used to detect the protein expression of osteogenic markers,collagenⅠ(COL-1),ALP,Runt-related transcription factor 2(RUNX2),and β-Catenin.Results The Micro-CT analysis revealed that at 2 months post-surgery,the fibrinogen-loaded collagen membrane group exhibited significantly higher levels of callus formation compared to the control group.ALP staining and Western blot results further indicated that fibrinogen promotes osteogenesis by upregulating the expression of osteogenic markers through the activation of the Wnt/β-Catenin signaling pathway.However,when PNU-74654(a Wnt/β-Catenin inhibitor)was utilized,the expression of osteogenic markers was significantly reduced(P＜0.05).Conclusion The fibrinogen-loaded collagen membrane can accelerate femur fracture healing in rats,and fibrinogen accomplishes this by promoting osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs)via the activation of the Wnt/β-Catenin signaling pathway.