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Abstract：
BACKGROUND: Repairation of bone defects remains a major clinical problem. Constructing bone tissue engineering containing growth factors, stem cells, and material scaffolds to repair bone defects has recently become a hot research topic. Nerve growth factor (NGF) can promote osteogenesis of bone marrow mesenchymal stem cells (BMSCs), but the low survival rate of the BMSCs during transplantation remains an unresolved issue. In this study, we investigated the therapeutic effect of BMSCs overexpression of NGF on bone defect by inhibiting pyroptosis.
METHODS: The relationship between the low survival rate and pyroptosis of BMSCs overexpressing NGF in localized inflammation of fractures was explored by detecting pyroptosis protein levels. Then, the NGF+/BMSCs-NSA-Sca bone tissue engineering was constructed by seeding BMSCs overexpressing NGF on the allograft bone scaffold and adding the pyroptosis inhibitor necrosulfonamide(NSA). The femoral condylar defect model in the Sprague-Dawley (SD) rat was studied by micro-CT, histological, WB and PCR analyses in vitro and in vivo to evaluate the regenerative effect of bone repair.
RESULTS: The pyroptosis that occurs in BMSCs overexpressing NGF is associated with the nerve growth factor receptor (P75NTR) during osteogenic differentiation. Furthermore, NSA can block pyroptosis in BMSCs overexpression NGF. Notably, the analyses using the critical-size femoral condylar defect model indicated that the NGF+/BMSCs-NSA-Sca group inhibited pyroptosis significantly and had higher osteogenesis in defects.
CONCLUSIONS: NGF+/BMSCs-NSA had strong osteogenic properties in repairing bone defects. Moreover, NGF+/BMSCs-NSA-Sca mixture developed in this study opens new horizons for developing novel tissue engineering constructs.
METHODS: The relationship between the low survival rate and pyroptosis of BMSCs overexpressing NGF in localized inflammation of fractures was explored by detecting pyroptosis protein levels. Then, the NGF+/BMSCs-NSA-Sca bone tissue engineering was constructed by seeding BMSCs overexpressing NGF on the allograft bone scaffold and adding the pyroptosis inhibitor necrosulfonamide(NSA). The femoral condylar defect model in the Sprague-Dawley (SD) rat was studied by micro-CT, histological, WB and PCR analyses in vitro and in vivo to evaluate the regenerative effect of bone repair.
RESULTS: The pyroptosis that occurs in BMSCs overexpressing NGF is associated with the nerve growth factor receptor (P75NTR) during osteogenic differentiation. Furthermore, NSA can block pyroptosis in BMSCs overexpression NGF. Notably, the analyses using the critical-size femoral condylar defect model indicated that the NGF+/BMSCs-NSA-Sca group inhibited pyroptosis significantly and had higher osteogenesis in defects.
CONCLUSIONS: NGF+/BMSCs-NSA had strong osteogenic properties in repairing bone defects. Moreover, NGF+/BMSCs-NSA-Sca mixture developed in this study opens new horizons for developing novel tissue engineering constructs.
摘要：
背景：修复骨缺损仍然是一个主要的临床问题。构建含生长因子的骨组织工程,干细胞,支架材料修复骨缺损已成为近期研究的热点。神经生长因子(NGF)促进骨髓间充质干细胞(BMSCs)成骨,但是BMSCs在移植过程中的低存活率仍然是一个未解决的问题。在这项研究中,我们研究了过表达NGF的BMSCs通过抑制焦凋亡对骨缺损的治疗作用。
方法：通过检测焦亡蛋白水平，探讨过表达NGF的BMSCs在骨折局部炎症中的低存活率与焦亡的关系。然后，NGF+/BMSCs-NSA-Sca骨组织工程是通过将过表达NGF的BMSCs接种在同种异体骨支架上，并添加焦亡抑制剂坏死磺酰胺（NSA）来构建的。通过显微CT研究了Sprague-Dawley（SD）大鼠的股骨髁缺损模型，组织学,体外和体内WB和PCR分析以评估骨修复的再生作用。
结果：过表达NGF的BMSCs在成骨分化过程中发生的焦亡与神经生长因子受体（P75NTR）有关。此外，NSA可以阻断过表达NGF的BMSCs的焦亡。值得注意的是,使用临界大小股骨髁部缺损模型进行的分析表明,NGF+/BMSCs-NSA-Sca组显著抑制了骨功能恢复,并在缺损中具有更高的成骨作用.
结论：NGF+/BMSCs-NSA在骨缺损修复中具有较强的成骨特性。此外，在这项研究中开发的NGF/BMSCs-NSA-Sca混合物为开发新的组织工程构建体开辟了新的视野。
方法：通过检测焦亡蛋白水平，探讨过表达NGF的BMSCs在骨折局部炎症中的低存活率与焦亡的关系。然后，NGF+/BMSCs-NSA-Sca骨组织工程是通过将过表达NGF的BMSCs接种在同种异体骨支架上，并添加焦亡抑制剂坏死磺酰胺（NSA）来构建的。通过显微CT研究了Sprague-Dawley（SD）大鼠的股骨髁缺损模型，组织学,体外和体内WB和PCR分析以评估骨修复的再生作用。
结果：过表达NGF的BMSCs在成骨分化过程中发生的焦亡与神经生长因子受体（P75NTR）有关。此外，NSA可以阻断过表达NGF的BMSCs的焦亡。值得注意的是,使用临界大小股骨髁部缺损模型进行的分析表明,NGF+/BMSCs-NSA-Sca组显著抑制了骨功能恢复,并在缺损中具有更高的成骨作用.
结论：NGF+/BMSCs-NSA在骨缺损修复中具有较强的成骨特性。此外，在这项研究中开发的NGF/BMSCs-NSA-Sca混合物为开发新的组织工程构建体开辟了新的视野。
