Objective: To determine the optimum staining condition of tea solutions on bovine incisors in vitro, by comparing the color stability of tooth surface of different concentrations of tea solutions and methods on bovine incisors in vitro.Methods:Twenty bovine incisors with color surface A1 were chosen, then randomly divided into 4 groups (n=10). Group 1: soaked with 2% tea solution continuously for 6 days; group 2: soaked with 2% tea solution for 6 days, but changed fresh tea solution everyday; group 3: soaked with 1% tea solution continuously for 6 days; group 4: soaked with 1% tea solution for 6 days but fresh tea solution changed every day. After 6 days of staining, the surface color (ΔE value)of all the samples were measured with crystal eye. After brushing 30 times with toothbrushes, the color of bovine incisors were measured again. Then the samples were soaked in artificial saliva at 37 ℃, and ΔE value was measured for 14 days.Results:After staining for 6 days, the ΔE values of the 2% tea solution groups were better than those of the 1% groups (20.21 vs. 16.44, 24.09 vs. 19.22, P<0.05);the groups with the same tea solution concentration, a better result was observed for the group soaked with daily fresh tea solution than for the group that experienced continuous staining (24.09 vs. 20.21, 19.22 vs. 16.44, P<0.05).Groups 1 and 2 were selected for subsequent brushing experiments. The color of both groups became lighter after brushing, and a better result was observed for the continuous staining group than for the group stained in daily fresh solution (3.06 vs. 9.51, P<0.05). The samples with better coloring effect soaked with 2% tea solution continuously for 6 days were put into artificial saliva for 14 days. There was not any significant change in coloring at theend of the first two days (1.51 vs. 1.51, P>0.05), and the color was visibly lighter after the third day (1.51 vs. 5.89, P<0.05), and no further significant change was observed until the 14th day (5.81 vs.5.89, P>0.05), which was darker coloring than that of the pre-staining group.Conclusion:Continuous staining on bovine incisors with 2% tea solution with subsequent soaking in artificial saliva resulted in consistent coloring from day 3 to day 14, and this method could be used as an ideal model for teeth staining in vitro.