International Journal of Nanomedicine (Jun 2025)
Multi-Focused Acoustic Radiation Force Impulse Modulation of Murine Hepatic Xenografts Enhances Nanoscale DOX@Lip Delivery and Therapeutic Effect
Abstract
Size Wu, Chengfang Wang Department of Ultrasound, The First Affiliated Hospital of Hainan Medical University, Haikou, Hainan, People’s Republic of ChinaCorrespondence: Size Wu, Department of Ultrasound, The First Affiliated Hospital of Hainan Medical University, No. 31, Longhua Road, Haikou, Hainan, 570102, People’s Republic of China, Email [email protected]: To investigate whether multi-focused acoustic radiation force impulse (MF-ARFI) applied to murine xenograft liver tumors prior to intravenous administration of doxorubicin-loaded PEGylated liposomes (DOX@Lip) can enhance drug delivery efficiency through modulating the enhanced permeation and retention effect of the tumor, reduce side effects, and improve antitumor effect.Materials and Methods: DOX@Lip and tumor-mimetic matrices were synthesized and characterized. Huh-7 cells and DOX@Lip were exposed to MF-ARFI and observed. MF-ARFI was applied to both tumor-mimetic matrices and saline with DOX@Lip to assess displacement effects. Subsequently, murine xenograft models were established and underwent MF-ARFI preconditioning before DOX@Lip injection. Tumor volume dynamics and body weight changes were longitudinally monitored. Terminal assessments included histopathology (H&E), apoptosis (TUNEL), and molecular profiling (BCL-2 and BAX by Western blot) of tumors and major organs.Results: There was no significant difference in the live/dead cell staining results between the Huh7 cells with and without MF-ARFI. There was no significant difference in cell apoptosis rates of Huh7 cells between DOX@Lip and DOX@Lip+MF-ARFI. MF-ARFI exposure induced measurable displacement of DOX@Lip in both tumor-mimetic matrices and saline. Mice receiving combined DOX@Lip and MF-ARFI treatment exhibited significantly attenuated tumor growth (p< 0.05) and slight weight loss, which were significantly different from DOX and DOX+MF-ARFI treatments. Cardiac histopathology revealed no significant differences in myocardial toxicity between DOX@Lip and DOX@Lip+MF-ARFI groups relative to PBS controls. Conversely, tumors from the DOX@Lip+MF-ARFI group demonstrated distinct histopathological alterations compared to other groups. TUNEL staining results indicated a relatively higher level of cell apoptosis in mice treated with DOX@Lip+MF-ARFI. Molecular analyses showed MF-ARFI pretreatment significantly reduced BCL-2 expression (p< 0.05) while elevating the BAX/BCL-2 ratio versus DOX@Lip monotherapy.Conclusion: Preconditioning xenograft tumors with MF-ARFI prior to DOX@Lip administration faciliates DOX@Lip delivery and significantly enhances antitumor effect while reducing cardiotoxicity. This combinatorial strategy demonstrates translational potential for optimizing liposomal chemotherapeutic delivery and effect.Plain language Summary: Doxorubicin (DOX) can be used for the therapy of some liver cancers ineligible for surgery. However, DOX has severe side effects in high concentration. Using liposomes to encapsulate DOX to form nanoscale DOX@Lip can control the release of DOX in the blood circulation and reduce side effects. DOX@Lip can pass the tumor capillary and accumulate in the tumor stroma and release sustainably. However, its release efficiency can be impacted by the tumor stroma status. If the tumor stroma becomes stasis due to necrotic tumor and other cellular debris after previous treatments, the subsequently administrated drug will be difficult to pass the capillaries to accumulate in the tumor stroma. Multi-focused acoustic radiation force impulse (MF-ARFI) is a kind of low-intensity ultrasound, with it irradiation of liver tumor, the structures of tumor stroma can occur push, pull, and displacement, facilitating the drug leak out the capillaries, promoting the waste substances in the stroma moving away. Therefore, using MF-ARFI before nanoscale drug administration can facilitate the drug delivery and distribution in the tumor. The results of this study showed that using MF-ARFI irradiated xenograft liver tumor of mice followed by intravenous infusion of DOX@Lip significantly enhanced anti-cancer effect and reduce side effects.Keywords: liver cancer, doxorubicin, liposomes, drug delivery, xenograft tumor, acoustic radiation force impulse, enhanced permeation and retention effect
