fig3

Figure 3. Intraosseous tumor area, apoptosis and proliferation. Tumor burden in bone was analyzed in histological sections of the left hind leg. Apoptotic tumor cells were stained using the TUNEL method and stained cells with apoptotic morphology were counted. Proliferation index was analyzed from Ki-67 stained sections. A: Representative images of HE + Orange G stained sections showing tumors in bone at sacrifice (day 25); B: representative images of the TUNEL stained sections; C: representative images of the Ki-67 stained sections; D: intraosseus tumor area did not differ between the groups; E: apoptotic tumor cells relative to tumor area were increased in both groups receiving DOX, but not in the ZOL group; F: proliferation index did not differ between the groups. Data are expressed as mean ± SD, n = 8 animals. Scale bars 1 mm (A); 50 μm (B and C). ***Significantly different from control group (P < 0.001); NS: non-significant; T: tumor; M: marrow; Tr.B.: trabecular bone; HE: hematoxylin-eosin; TUNEL: terminal deoxynucleoitidyl transferase dUTP nick end labeling; DOX: doxorubicin; ZOL: zoledronic acid