Kuang Zhiping, Dai Guangming, Wan Ruijie, Zhang Dongli, Zhao Chen, Chen Cheng, Li Jidong, Gu Hongchen, Huang Wei
Department of Orthopedic Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, PR China.
Department of Orthopaedic Surgery, Chongqing Traditional Chinese Medicine Hospital, Chongqing, 400011, PR China.
Genes Dis. 2019 Oct 5;8(2):193-202. doi: 10.1016/j.gendis.2019.09.014. eCollection 2021 Mar.
Lev/MSNs/n-HA/PU has been proved to be a novel scaffold material to treat bone defect caused by chronic osteomyelitis. We have previously identified that this material can effectively treat chronic osteomyelitis caused by . However, the potential mechanisms of antibacterial and osteogenic induction properties remain unclear. Thus, for osteogenesis property, immunohistochemistry, PCR, and Western blot were performed to detect the expression of osteogenic markers. Furthermore, flow cytometry and TUNEL were applied to analyze MC3T3-E1 proliferation and apoptosis. For antibacterial property, the material was co-cultivated with bacteria, bacterial colony forming units was counted and the release time of the effective levofloxacin was assayed by agar disc-diffusion test. Moreover, scanning electron microscope was applied to observe adhesion of bacteria. In terms of osteogenic induction, we found BMSCs adherently grew more prominently on Lev/MSNs/n-HA/PU. Lev/MSNs/n-HA/PU also enhanced the expression of osteogenic markers including OCN and COL1α1, as well as effectively promoted the transition from G1 phase to G2 phase. Furthermore, Lev/MSNs/n-HA/PU could reduce apoptosis of MC3T3-E1. Besides, both Lev/MSNs/n-HA/PU and n-HA/PU materials could inhibit bacterial colonies, while Lev/MSNs/n-HA/PU possessed a stronger antibacterial activities, and lower bacterial adhesion than n-HA/PU. These results illustrated that Lev/MSNs/n-HA/PU composite scaffold possess favorable compatibility , which induce osteogenic differentiation of MSCs, promote proliferation and differentiation of MC3T3-E1, and inhibit apoptosis. Moreover, clear antibacterial effect of Lev/MSNs/n-HA/PU was also observed. In summary, this study replenishes the potential of Lev/MSNs/n-HA/PU composite scaffold possess dual functions of anti-infection and enhanced osteogenesis for future clinical application.
左旋咪唑/介孔二氧化硅纳米粒子/纳米羟基磷灰石/聚氨酯已被证明是一种治疗慢性骨髓炎所致骨缺损的新型支架材料。我们之前已经确定这种材料可以有效治疗由……引起的慢性骨髓炎。然而，其抗菌和成骨诱导特性的潜在机制仍不清楚。因此，对于成骨特性，进行了免疫组织化学、聚合酶链反应和蛋白质免疫印迹法来检测成骨标志物的表达。此外，应用流式细胞术和末端脱氧核苷酸转移酶介导的缺口末端标记法来分析MC3T3-E1细胞的增殖和凋亡。对于抗菌特性，将该材料与细菌共培养，计数细菌菌落形成单位，并通过琼脂扩散试验测定有效左氧氟沙星的释放时间。此外，应用扫描电子显微镜观察细菌的黏附情况。在成骨诱导方面，我们发现骨髓间充质干细胞在左旋咪唑/介孔二氧化硅纳米粒子/纳米羟基磷灰石/聚氨酯上的贴壁生长更为显著。左旋咪唑/介孔二氧化硅纳米粒子/纳米羟基磷灰石/聚氨酯还增强了包括骨钙素和I型胶原蛋白α1在内的成骨标志物的表达，并有效促进了细胞从G1期向G2期的转变。此外，左旋咪唑/介孔二氧化硅纳米粒子/纳米羟基磷灰石/聚氨酯可以减少MC3T3-E1细胞的凋亡。此外，左旋咪唑/介孔二氧化硅纳米粒子/纳米羟基磷灰石/聚氨酯和纳米羟基磷灰石/聚氨酯材料都可以抑制细菌菌落，而左旋咪唑/介孔二氧化硅纳米粒子/纳米羟基磷灰石/聚氨酯具有更强的抗菌活性，且细菌黏附比纳米羟基磷灰石/聚氨酯更低。这些结果表明，左旋咪唑/介孔二氧化硅纳米粒子/纳米羟基磷灰石/聚氨酯复合支架具有良好 的生物相容性，可诱导间充质干细胞的成骨分化，促进MC3T3-E1细胞的增殖和分化，并抑制细胞凋亡。此外，还观察到左旋咪唑/介孔二氧化硅纳米粒子/纳米羟基磷灰石/聚氨酯具有明显的抗菌效果。总之，本研究补充了左旋咪唑/介孔二氧化硅纳米粒子/纳米羟基磷灰石/聚氨酯复合支架具有抗感染和增强成骨双重功能的潜力，以供未来临床应用。
