02 ± 0.21), suggesting complete recovery of tumoral activities at the later stage of treatment (Figure 6A, B). However, the RSI of BLI in group D dropped 3 days after treatment as in group C and exhibited minimal recovery until day 14 post-treatment (0.31 ± 0.20) (Figure 6A, B).The Mann–Whitney test performed for the BLI values at day 14 post-treatment revealed that the RSI of BLI in group D was significantly lower than the other

groups (all p-values < 0.05) (Figure 6C). The exact p-values obtained between groups are summarized in Table 1. No mouse exhibited signs of debilitation in any of the groups through the follow-up period. Figure 6 Bioluminescence imaging (BLI). A) Representative BLI obtained in each group by the IVIS lumina II (PerkinElmer, Waltham, MA). B) Relative signal intensity [RSI] of BLI over the follow-up selleck screening library period. C) A graph demonstrated the relative signal intensity [RSI] of BLI at 14 days after treatment (*P < 0.05, compared to group A). Histopathological findings TUNEL assay of the tumor tissues obtained at day 14 by revealed that the apoptosis/necrosis rate in group D was DNA Damage inhibitor higher (39.0 ± 13.2%) than group A (11.52 ± 3.10%), B (25.4 ± 3.36%), and C (23.0 ± 7.68%) (Figure 7). Therefore, the Resovist/doxorubicin complex showed significantly more cell death than doxorubicin or Resovist monotherapy (all p-values < 0.05).The exact p-values obtained between

groups are summarized in Table 1. Prussian blue staining of the consecutive section demonstrated Abiraterone multiple iron deposits within the tumor tissues in groups C and D (Figure 8). Figure 7 Terminal deoxynucleotidyl transferase-mediated nick end NF-��B inhibitor labeling (TUNEL) assays to measure apoptotic cell death by light microscopy. A) TUNEL-positive (brown color) cells with apoptotic morphology were observed

in all groups (x200). B) A graph demonstrating the apoptosis/necrosis rates in all groups by image J software (*P < 0.05, compared to group A). Figure 8 Histopathological analyses of the tumor tissues by light microscopy. Hematoxylin and eosin staining (left), Prussian blue staining (middle), and TUNEL staining (right) of a tumor treated with the Resovist/doxorubicin complex (x100). Doxorubicin fluorescence microscopic findings On fluorescence microscopic examination, group D exhibited higher fluorescence intensity from doxorubicin in the tumor tissues, which significantly overlapped the area with the iron particles. By contrast, group B exhibited minimal fluorescence from doxorubicin (Figure 9). This result suggests that the Resovist/doxorubicin complex could release doxorubicin into the tumor tissues in a controlled manner for a longer period than free doxorubicin and allowed persistent drug accumulation. Figure 9 Fluorescence microscopy images. Representative fluorescence images of group B (left), group C (middle), and group D (right).