目的探讨不同浓度钛合金颗粒对体外培养的小鼠成骨细胞的细胞外信号调控激酶1/2(ERK1/2)蛋白磷酸化水平的影响。方法①实验设空白对照组(control)和钛合金颗粒干预组(0.1g/L),每组又分为0、0.5、1、3和6h5个亚组。分别在培养第0、0.5、1、3和6h5个时点,采用WesternBlot法检测不同时间点ERK1/2磷酸化水平。②实验设空白对照组(control),钛合金颗粒干预A组(0.01g/L)、B组(0.1g/L)和C组(1g/L)。在培养第0.5h,采用WesternBlot法检测不同钛合金颗粒浓度作用下成骨细胞ERK1/2磷酸化水平。结果在第0.5、1、6h时点,钛合金颗粒干预组(0.1g/L)ERK1/2磷酸化水平均较对照组高,差异有统计学意义(P〈0.01),其中0.5h亚组成骨细胞ERK1/2蛋白磷酸化水平最高;但在第3h时点,2组ERK1/2磷酸化水平的差异无统计学意义(P〉0.05)。不同浓度钛合金颗粒共培养的结果显示,不同浓度钛合金颗粒组成骨细胞的ERK1/2磷酸化水平均高于对照组,差异有统计学意义(P〈0.05或P〈0.01)。结论不同浓度钛合金颗粒可促进成骨细胞ERK蛋白磷酸化水平,激活ERK信号通路。
Objective To investigate the effects of different concentrations of titanium alloy particles on the phos- phorylated ERK1/2 level in mouse osteoblasts in vitro. Methods The osteoblasts were divided into a control group and an experimental group treated with 0.1 g/L titanium alloy particles. According to the culture time (0, 0.5, 1, 3 and 6 h) , each group was divided into five sub - groups. The level of phosphorylated ERK - 1/2 was detected by Western blot- ting at various time points. Meanwhile, the osteoblasts were divided into four groups: a control group, and three experi- mental groups (0. 01 g/L, 0.1 g/L and 1 g/L). The level of phosphorylated ERK -1/2 was detected by Western blot- ting after treatment with different concentrations of titanium alloy particles. Results The level of phosphorylated ERK1/ 2 was significantly higher in the experimental group (0.1 g/L) than that in the control group at 0.5, 1, and 6 h ( P 〈 0. 01 ), which reached the peak at 0.5 h. But no statistical significance was found between the two groups at 3 h ( P 〉 0.05 ). Further studies demonstrated that a higher level of phosphorylated ERK1/2 in each experimental group using dif- ferent concentrations of titanium alloy particle groups, compared with the control group (P 〈 0.05 or P 〈 0.01 ). Con- clusions Different concentrations of titanium alloy particles can stimulate the phosphorylation of ERK1/2, and activate the ERK signal pathways.