Fig. 4
Semiquantitative analysis of calcification. Group 1 (control): untreated tissues; group 2: tissues fixed with glutaraldehyde (GA) for 3 min; group 3: tissues fixed with GA for 3 min and post-treated with ethanol for 10 min; group 4: tissues fixed with GA for 3 min and post-treated with ethanol for 20 min; group 5: tissues fixed with GA for 3 min and post-treated with ethanol for 30 min. Semiquantitative analysis of calcification revealed the calcified areas in the explants of groups 1, 2, 3, 4, and 5 to be 0.02 ± 0.04, 0.22 ± 0.15, 0.10 ± 0.07, 0.07 ± 0.03, and 0.08 ± 0.03 mm2, respectively. Calcification of explants from groups 1 (P = 0.0014), 4 (P = 0.0014), and 5 (P = 0.0031) was significantly lower than that of explants from group 2. Group 3 (P = 0.0503) showed slightly decreased calcification although the decrease was not statistically significant. Calcified areas are displayed as a bow-and-whiskers plot with median (thick line), 25th to 75th percentiles (box), 10th to 90th percentiles (whiskers), and outliers (circles). P-values were calculated using the Kruskal–Wallis test