Figure 1.
Characterization of the DWC model.
(A) Filling of tumor blood vessels after i.o. injection of FD and increase of fluorescence intensity in the tumor tissue in control mice. Mean fluorescence intensities were expressed as a percentage of the maximum mean fluorescence intensity (Imax) reached in the observation period. The first image in the series was acquired ∼10 s after the i.o. injection of FD. (B) The timeline of the protocol used in the experiments.
Figure 2.
Imaging of the leakage of FD from tumor blood vessels into the tumor tissue.
Tumor blood vessels were visualized by fluorescence microscopy at 80×magnification. Control – mice without treatment, Bleomycin – mice treated with bleomycin only, EP – mice treated with EP, ECT – mice treated with ECT. Tumors are marked with a dashed line. Scale bar is 500 µm.
Figure 3.
Quantification of FD leakage kinetics from tumor blood vessels.
Relative mean fluorescence intensity changes as a function of time in the tumor tissue, outside the tumor blood vessels. Control – mice without treatment, Bleomycin – mice treated with bleomycin only, EP – mice treated with EP, ECT – mice treated with ECT. n = 3–5. **p<0.05 compared to all other groups, *p<0.05 compared to control and EP groups. Error bars indicate SEM.
Figure 4.
Illustration of a “vascular lock” and re-perfusion of tumor blood vessels after EP and ECT.
Tumor blood vessels were visualized by fluorescence microscopy at 80×magnification. Control – mice without treatment, Bleomycin – mice treated with bleomycin only, EP – mice treated with EP, ECT – mice treated with ECT. (A) FD was injected i.o. at different times after the therapy (1, 10, 30, 60 min), and images were taken at designated times. Scale bar is 500 µm.
Figure 5.
Illustration of a “vascular lock” and re-perfusion of tumor blood vessels in the first hour and 4–24 h after EP and ECT.
Tumor blood vessels were visualized by fluorescence microscopy. Control – mice without treatment, EP – mice treated with EP, ECT – mice treated with ECT. (A) FD was injected i.o. 1 min after EP, and images were acquired at 20×magnification at designated times. Arrows indicate the position of the tumor. Scale bar is 2 mm. (B) RhD was injected i.o. at 4, 8 and 24 h after the therapy, and images were acquired 5 min after the injection at 80×magnification. The images are representative of different tumors. Tumors are marked with a dashed line. Scale bar is 500 µm.
Figure 6.
Timeline of the decrease in FVD and increase in DV after EP and ECT.
Control – mice without treatment, Bleomycin – mice treated with bleomycin only, EP – mice treated with EP, ECT – mice treated with ECT. (A) The changes in FVD within the tumors are presented as a function of time. (B) The changes of DV within the tumors are presented as a function of time. n = 3–6. **p<0.05 compared to all other groups, *p<0.05 compared to control and bleomycin groups. Error bars indicate SEM.
Figure 7.
Illustration of FD leakage from blood vessels surrounding the tumor and their constriction after EP and ECT.
Images were acquired at 20×magnification. Control – mice without treatment, Bleomycin – mice treated with bleomycin only, EP – mice treated with EP, ECT – mice treated with ECT. (A) Illustration of FD leakage from blood vessels surrounding the tumor, when FD was injected i.o. before the therapy. Arrows indicate the position of the tumor. Scale bar is 2 mm. (B) Illustration of the constriction of blood vessels surrounding the tumor visualized by FD. The enlarged sections of images were taken at 20×magnification. Scale bar is 500 µm.