The effect of dual‐frequency ultrasound waves on B16F10 melanoma cells: Sonodynamic therapy using nanoliposomes containing methylene blue

Abstract

Background We investigated the effect of dual‐frequency sonication on the viability of B16F10 melanoma cells in the presence of methylene blue (MB) encapsulated in nanoliposomes. Methods Treatment protocols were studied: sonication groups (40 kHz, 1 MHz and dual‐frequency), the same sonication groups with nanoliposomes containing MB, MB free and nanoliposomes containing MB groups, and so sham and control groups. The nanoliposomes were prepared by the lipid film hydration method. The cell viability of the different treatment groups was evaluated by the MTT assay. Results The dual‐frequency protocols caused higher viability losses compared to the kHz and MHz sonications (P < .05). In presence of the nanoliposomes containing MB, dual frequency led to 6% and 3% viability for 600 and 1200 seconds, respectively, while the corresponding values were 10% and 4% for the 40 kHz protocols and 22% and 9% for the 1 MHz, as compared to the control group (100%). The result of KI dosimetry showed that the cavitation activity of the dual‐frequency protocol was about 1.23, as compared to sonication at 40 kHz and 1 MHz. Conclusion Enhancement of inertial cavitation induction by dual‐frequency sonication may be the primary effective mechanism, which causes increased sonochemical processes and drug release from nanocarriers.

Multi-frequency therapeutic ultrasound: A review

“…Different implementations of multi-frequency ultrasound are also summarized, including single-element transducer, double transducers, triple transducers and phased array transducers, each with its own specialty and favored for particular tasks. Benefited from lesion size control, cavitation enhancement, low power needs and multi-foci treatment, this technique offers a wide range of applications, including: sonothrombolysis, tissue (tumor) ablation, sonodynamic treatment, sonochemical treatment, temperature rise, cell differentiation, BBB opening, transdermal drug delivery, sub-harmonic imaging enhancement and split-focus treatment [2] , [6] , [7] , [11] , [12] , [13] , [14] , [15] , [16] , [17] , [18] , [22] , [23] , [25] , [28] , [30] , [32] , [33] , [36] , [50] , [51] , [53] , [54] , [55] , [60] , [68] , [69] , [70] , [71] , [72] , [73] , [74] , [75] , [76] , [77] , [78] , [79] , [80] , [81] , [82] , [83] , [84] , [85] , [86] , [23] , [75] . Moreover, this review discusses about the decisive parameters for multi-frequency treatment and suggests guidelines for parameter selection.…”

“…Over the years of development, multi-frequency therapeutic ultrasound is widely used in a series of medical applications, including sonothrombolysis [14] , [36] , tissue (tumor) ablation [53] , [68] , sonodynamic treatment [69] , [70] , sonochemical treatment [71] , [72] , temperature rise [2] , [73] , cell differentiation [16] , [51] , BBB opening [66] , [67] , transdermal drug delivery [15] , [74] and sub-harmonic imaging [50] , etc. Moreover, as a side function, the multi-transducers and phased-array transducers further allow multi-task treatments [23] , [75] .…”

“…The growth was reduced by 46 % compared to the control group. Several other sonodynamic treatment enhancement were investigated using dual-frequency sonication, and most of which focused on tumor treatment [69] , [70] , [80] . In this type of application, the focused ultrasound plays a role as a triggering tool for medical agents instead of direct influence.…”

Synergistic Piezo-Catalytic Inactivation of Bacteria by Dual-Frequency Ultrasound (120 + 1700 kHz) Using Persulfate and ZnO Nano- and Microparticles

“…The log reductions in single-frequency mode were obtained for the same exposure periods, which were needed for achieving a 5-log reduction in DFUS mode (Table 1). A fixed time for evaluating a synergy of DFUS was also previously used by Adelnia et al (2020) [43]. Given that approach, the synergistic coefficient was determined relative to the 5-log reduction times for the DFUS process (1):…”

Combined use of microbubbles of various sizes and single‐transducer dual‐frequency ultrasound for safe and efficient inner ear drug delivery

“…A dual‐frequency transdermal delivery system has also been proposed, with two US frequencies (20 kHz and 1 MHz) generated separately from various transducers to enhance transdermal delivery efficiency, and it has outperformed a single‐frequency system using 20 kHz 18 . Different inventions, including intravascular sonothrombolysis, 19 sonodynamic therapy on melanoma cells, 20 and the induction of cancer stem cells for differentiation therapy for hepatocellular carcinoma, 21 have capitalized on the benefits of using dual‐frequency USMBs cavitation instead of single‐frequency US. The critical experimental observation is that multifrequency US can lower the inertial cavitation threshold, thereby improving the power efficiency 22 .…”