Two-way ANOVA with Bonferroni
post-test analysis was applied to address the following three questions: i) does age have the same effect at all values of treatment (interaction), ii) does age affect the result, and iii) does treatment affect the result. Where this type of analysis indicated significant differences, additional comparisons were made employing unpaired t-test. Additionally, 2-way correlations between spectroscopically determined pyd/divalent collagen cross-link ratio, structural and mechanical properties were explored using Spearman’s test. Significance was assigned to p < 0.05. The animals did not show any effect of the diet during the treatment period. There was no statistical difference in weight between the control and β-APN-treated animal groups at either time point although both groups gained weight over the CHIR-99021 in vitro two week period (data not shown). Two-way ANOVA of qBEI measurements also showed no statistically significant differences between the animal groups at either time point in any of
the four outcomes monitored (Table 1), with the exception of CaPeak which was dependent on animal age but not treatment. Biochemical analysis indicated that changes in DHLNL were dependent on both animal age and treatment, while PYD and DPD were dependent only on treatment. The calculated PYD/divalent cross-link ratio was dependent on both animal age and treatment (Table 2). Further comparisons using unpaired t-tests showed significant differences in DHLNL, between control Cobimetinib datasheet and β-APN-treated animals at the 4 week time point, and a time-dependent difference in the β-APN treated animals at 2 and 4 weeks (Fig. 1a).
The concentration of the trivalent cross-link PYD was significantly different between the control and β-APN treated animals at the 4-week time point (Fig. 1b). The other trivalent collagen cross-link, DPD, was significantly different between control and treated animals at both time points (Fig. 1c). The calculated ratio between PYD/DHLNL was increased in both groups as a function of time, and was elevated in the 4 week click here treated animal group compared to the 2 week treated and the 4 week control groups (Fig. 1d). Two-way ANOVA analysis (Table 3) of structural parameters determined by μ-CT analysis of vertebral bone revealed no interaction between factor age and treatment. Trabecular BV/TV and TRI-SMI were influenced only by treatment, trabecular thickness by age and treatment, and trabecular DIM-Z by age only. Additionally, cortical thickness was influenced by both age and treatment. Further statistical analysis employing unpaired t-tests a significantly lower BV/TV in the treated animals at 4 weeks compared to the corresponding controls (Fig. 2a). Differences in Structural Model Index (TRI-SMI) were also observed with age and in treated animals for 4 weeks compared to corresponding controls (Fig. 2b).