目的:本研究涉及低强度脉冲超声(LIPUS)和铁(Fe3)离子处理的成骨细胞的增殖和分化,分别。同时评价了LIPUS和Fe3+离子对成骨细胞增殖和分化的生物学效应。
方法:用培养基将MC3T3-E1细胞接种在6孔板中,其中含有不同浓度的Fe3(分别为0、100、200、300、400、500、600和700μgL-1)。LIPUS处理以每天80mWcm-2的强度在板的底部持续20分钟。
结果:活力结果显示,400μgL-1Fe3+离子的剂量在促进细胞培养成骨增殖方面效果最好。碱性磷酸酶染色和矿化结果表明,LIPUS和Fe3离子促进了成骨细胞的分化。荧光染色结果显示,LIPUS中的细胞核数量,Fe3+和LIPUS-Fe基团增加37.20%,55.81%和89.76%,分别。迁移数据表明,LIPUS和Fe3+增加了迁移和增殖率,蛋白表达结果表明,LIPUS和Fe3+可以增加Wnt的表达,β-连环蛋白,和Runx2,从而促进正常的骨再生和发育。
结论:LIPUS(1.5MHz,80mWcm-2)和Fe3+与单因素处理(LIPUS和Fe3+离子刺激,分别)。本研究为LIPUS响应性生物材料在骨组织修复和再生中的应用奠定了基础。
OBJECTIVE: This study involved the proliferation and differentiation of osteoblasts treated with low-intensity pulsed ultrasound (LIPUS) and iron (Fe3+) ions, respectively. The biological effects of LIPUS and Fe3+ ions on the proliferation and differentiation of osteoblasts were also evaluated.
METHODS: MC3T3-E1 cells were seeded in six-well plates with the medium, which contained different concentrations of Fe3+ (0, 100, 200, 300, 400, 500, 600 and 700 μg L-1, respectively). LIPUS treatment was directed at the bottom of the plate for 20 min at an intensity of 80 mW cm-2 every day.
RESULTS: Viability results showed that a dose of 400 μg L-1 Fe3+ ions had the best effect at promoting osteogenic proliferation in cell culture. The results of alkaline phosphatase staining and mineralization indicated that the differentiation of osteoblasts was promoted by LIPUS and Fe3+ ions. Fluorescence staining results showed that the number of cell nuclei in the LIPUS, Fe3+ and LIPUS-Fe groups increased by 37.20%, 55.81% and 89.76%, respectively. Migration data indicated that migration and proliferation rates were increased by LIPUS and Fe3+, and the results of protein expression indicated that LIPUS and Fe3+ may increase the expression of Wnt, β-catenin, and Runx2, hence promoting normal bone regeneration and development.
CONCLUSIONS: The combination of LIPUS (1.5 MHz, 80 mW cm-2) and Fe3+ accelerates the proliferation and differentiation of osteoblasts significantly compared with single-factor treatment (stimulated by LIPUS and Fe3+ ions, respectively). This study could establish a foundation for LIPUS-responsive biomaterials in the repair and regeneration of bone tissues.