循环轴向压缩应力增强基质依赖型组织工程骨骨再生能力的研究  

Cyclic axial compression stress promotes the bone regeneration ability of matrix-based tissue engineering bone

作　　者：梁万元 胡文辉 丁海滨 李建美 董世武 Liang Wanyuan;Hu Wenhui;Ding Haibin;Li Jianmei;Dong Shiwu(Department of Biomedical Materials Science,National and Regional Engineering Laboratory of Tissue Engineering,School of Biomedical Engineering and Imaging Medicine,Army Medical University(Third Military Medical University),Chongqing,400038,China)

机构地区：[1]陆军军医大学生物医学工程与影像医学系生物医学材料学教研室,组织工程发改委国家-地方联合工程实验室,重庆400038

出　　处：《生物骨科材料与临床研究》2022年第5期1-8,共8页Orthopaedic Biomechanics Materials and Clinical Study

基　　金：全军后勤保障研究计划重点项目(AWS17J004-02-06);陆军军医大学军事医学预研基金(2018XYY05)。

摘　　要：目的研究循环轴向压缩应力(CACS)对基质依赖型组织工程骨(M-TEB)骨再生能力的影响。方法首先,在构建静态M-TEB过程中加载CACS,得到动态M-TEB;然后,对各组M-TEB进行表征,并构建动物模型来评价其骨再生能力;最后,通过转录组测序探究CACS对骨髓间充质干细胞(BMSCs)基因表达的影响,并研究M-TEB来源条件培养基对内皮祖细胞(EPCs)迁移、增殖和对BMSCs成骨分化的影响。结果①动态M-TEB组在骨缺损区有更多新骨生成,且骨体积分数和骨密度均显著优于假手术组和静态M-TEB组(P均<0.01);②前30个有显著差异(q<0.01)的基因本体论(GO)术语主要涉及MAPK通路、细胞凋亡和血管生成等,且差异基因VEGFA在这些GO术语中出现频次最高(28次);③相比静态M-TEB来源条件培养基,动态M-TEB来源条件培养基具有更强的促进EPCs迁移、增殖和BMSCs成骨分化的作用(P均<0.05),阻断实验证实VEGFA在其中发挥重要功能。结论CACS促进BMSCs表达和分泌VEGFA,提高动态M-TEB中VEGFA浓度,最终增强其在大鼠股骨缺损模型中的骨再生能力。Objective To investigate the effect of cyclic axial compression stress(CACS) on the bone regeneration ability of matrix-based tissue engineering bone(M-TEB). Methods The construction process of M-TEB switched from static to dynamic after CACS loading. M-TEB was characterized by SEM and DNA quantification, and its bone regeneration ability was evaluated in the rat femoral defect model. RNA-seq was conducted to investigate differential gene expression of CACS-loaded BMSCs, and we explored the effects of conditioned medium(CM) extracting from M-TEB on migration and proliferation of endothelial progenitor cells(EPCs) and osteogenic differentiation of BMSCs. Results The dynamic M-TEB group had a greater amount of new bone formation in the bone defect area, and bone volume fraction and bone mineral density were significantly higher than those in the sham and static M-TEB groups(all P<0.01). GO enrichment analysis demonstrated that the top 30 terms mainly included MAPK pathway, apoptosis and angiogenesis(q<0.01), while VEGFA appeared the most frequently(28 times) among these terms. The CM from the dynamic M-TEB group promoted the migration and proliferation of EPCs, and the osteogenic differentiation of BMSCs(all P<0.05), compared with the static group. Furthermore, the blocking experiment confirmed the important existence of VEGFA in the CM. Conclusion CACS promoted the secretion of VEGFA from BMSCs into the extracellular matrix, increased the concentration of VEGFA in the dynamic M-TEB, and finally enhanced its bone regeneration ability in the femoral defect of rat model.

关 键 词：基质依赖型组织工程骨 循环轴向压缩应力 骨髓间充质干细胞 血管内皮生长因子A 

分 类 号：R318[医药卫生—生物医学工程]