Image analysis of microarchitectural changes to scaffolds with 6% MDI crosslinking in the villus layer and 2% MDI in the base layer during in vitro degradation. (A) SEM of the scaffolds at each time point in media alone, seeded with enteroids, and in digestive media (scale bars = 250 μm). The overall dimensions and villus architecture are maintained. (B–E) The quantification of the image analysis. For thickness, there was a statistically significant variation in time (p = 0.0012), but on post-hoc comparison of each time to baseline within each group, there were no statistical differences. There were no differences for villus height. There were significant differences in villus tip in both the time and group variables (p < 0.0001 and p = 0.0113, respectively). On multiple comparisons, there was a difference in the media group between 0 and 6 weeks (p = 0.0007), in the enteroid group between baseline and 2, 4, and 6 weeks (p = 0.0056, 0.0318, and <0.0001, respectively), and in the digest group, there were differences for each time point (2, 4, and 6 weeks) compared to baseline (p = 0.0004, 0.004, and <0.0001, respectively). Regardless of the statistical differences, these changes in size were small and in many cases, there were increases in dimension, which more likely represents either measurement error or some other artifact. @, #, and * indicate statistical significance. @ represents differences between time points for enteroids, # represents differences between time points for digest group, and * indicates difference in time points for media group. Color images are available online.