Fig 1.
Synthesis of microbubble (MB), doxorubicin binding human serum albumin nanoparticle (DOX-NPs) and DOX-NPs conjugated microbubble complex (DOX-MAC).
Fig 2.
Experimental design for the animal study, showing the timeline of each group for N1-S1 cell inoculation, doxorubicin-loaded microbubble-albumin nanoparticle complex (DOX-MAC) with/without sorafenib and doxorubicin (DOX) therapy, microbubble (MB) CEUS imaging, and animal sacrifice for tumor harvest as described in the “Materials and Methods” section.
Fig 3.
Serial changes of the relative tumor growth and rat weight.
(A) The tumor volume on a specific day divided by the baseline tumor volume showed the most delayed tumor growth at two weeks in the G1 group (P = 0.008). (B) The rat weight on a specific day divided by the baseline weight showed the slow increasing tendency of the rat two weeks in the G1 group. However, there was no significant difference between the control group and the G1 group (P > 0.05).
Table 1.
Serial changes of the tumor volume.
Table 2.
Serial changes of the rat weight.
Fig 4.
Serial changes of the perfusion parameters.
(A) The normalized peak enhancement (nPE) difference between the G1 group and the G5 group increased from baseline to two weeks (P = 0.010) (B) The normalized wash-in area under the curve (nWiAUC) difference between the G1 group and the G5 group increased from baseline to two weeks (P = 0.022). (C) There was no difference in the normalized wash-out area under the curve (nWoAUC) value between the G1 and the G5 group (G1 vs. G5, P = 0.05).
Table 3.
Serial changes of the normalized contrast-enhanced perfusion parameters.
Fig 5.
Representative cases of the G1 and G4 groups.
(A) The contrast-enhanced ultrasound showed the hyper-vascular tumor in the left lobe of the liver (upper, arrows). The mass showed a decrease of vascularity after combination treatment of sorafenib (SOR) and doxorubicin-loaded microbubble-albumin nanoparticle complex (DOX-MAC) (lower, arrows). (B) One of the G5 group showed the small hyper-vascular tumor in the left lobe of the liver (upper, arrows). The mass showed an increase of tumor volume with hyper-vascularity without any treatment (lower, arrows). (C) The number of brown-colored stained apoptotic cells on TUNEL staining (×400) was a subtle increase in the G1 group (upper, arrowheads) than the G5 group (lower, arrowheads) without statistical significance. (D) The number of brown-colored stained endothelial cells on CD31 staining (×200) decreased in the G1 group (upper, arrowheads) than the G5 group (lower, arrowheads).
Table 4.
Histologic parameters.