负载纤维蛋白原的胶原膜对大鼠股骨骨折愈合的作用及机制研究
The Effect and Mechanism of Collagen Membranes Loaded With Fibrinogen on the Healing of Femur Fractures in Rats
杨平 1雷智杰 2于江林 2邱旭升 2陈一心 3戚晓阳3
作者信息
- 1. 南京中医药大学鼓楼临床医学院骨科,江苏南京 210000
- 2. 江苏大学鼓楼临床医学院骨科,江苏南京 210000
- 3. 南京中医药大学鼓楼临床医学院骨科,江苏南京 210000;南京大学医学院附属鼓楼医院骨科,江苏南京 210000
- 折叠
摘要
目的 研究负载纤维蛋白原的胶原膜对大鼠股骨骨折愈合的影响及机制.方法 将大鼠股骨骨折模型随机分为三组,分别为对照组、胶原膜组、负载纤维蛋白原胶原膜组.术后第8周取患侧股骨行Micro-CT分析骨痂形成情况.大鼠原代骨髓间充质干细胞(bone marrow mesenchymal stem cell,BMSC)分为成骨对照组、纤维蛋白原+成骨诱导组、纤维蛋白原+PNU-74654+成骨诱导组.BCIP/NBT染色法检测BMSC的碱性磷酸酶(alkaline phosphatases,ALP)水平,Wsetern blot分别检测成骨指标Ⅰ型胶原蛋白(collagen Ⅰ,COL-1)、ALP、Runt相关转录因子 2(Runt-related transcription factor 2,RUNX2)、β-Catenin 的蛋白表达.结果 Micro-CT 显示术后 2 个月大鼠股骨骨折端的骨痂形成量,负载纤维蛋白原胶原膜组显著多于胶原膜组和对照组.ALP染色和Western blot结果显示纤维蛋白原能够促进成骨表,且纤维蛋白原通过激活Wnt/β-Catenin信号通路上调成骨指标的表达,而使用PNU-74654(Wnt/β-Catenin通路抑制剂)后成骨指标表达显著下降(P＜0.05).结论 负载纤维蛋白原的胶原膜能加速骨折愈合.纤维蛋白原通过激活Wnt/β-Catenin信号通路能促进BMSC成骨分化.
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.
关键词
纤维蛋白原/胶原膜/骨折愈合/成骨分化Key words
fibrinogen/collagen membrane/fracture healing/osteogenic differentiation引用本文复制引用
出版年
2024
NETL
NSTL
万方数据