RESUMO
Ultrasound localization microscopy (ULM) enables the creation of super-resolved images and velocity maps by localizing and tracking microbubble contrast agents through a vascular network over thousands of frames of ultrafast plane wave images. However, a significant challenge lies in developing ultrasound-compatible microvasculature phantoms to investigate microbubble flow and distribution in controlled environments. In this study, we introduce a new class of gelatin-based microfluidic-inspired phantoms uniquely tailored for ULM studies. These devices allow for the creation of complex and reproducible microvascular networks featuring channel diameters as small as 100 µm. Our experiments focused on microbubble behavior under ULM conditions within bifurcating and converging vessel phantoms. We evaluated the impact of bifurcation angles (25, 45, and 55°) and flow rates (0.01, 0.02, and 0.03 mL/min) on the acquisition time of branching channels. Additionally, we explored the saturation time effect of narrow channels branching off larger ones. Significantly longer acquisition times were observed for the narrow vessels, with an average increase of 72% when a 100 µm channel branched off from a 300 µm channel and an average increase of 90% for a 200 µm channel branching off from a 500 µm channel. The robustness of our fabrication method is demonstrated through the creation of two trifurcating microfluidic phantoms, including one that converges back into a single channel, a configuration that cannot be achieved through traditional methods. This new class of ULM phantoms serves as a versatile platform for noninvasively studying complex flow patterns using ultrasound imaging, unlocking new possibilities for in vitro microvasculature research and imaging optimization.
RESUMO
Scaling down the size of microbubble contrast agents to the nanometer level holds the promise for noninvasive cancer therapy. However, the small size of nanobubbles limits the obtained bioeffects as a result of ultrasound cavitation, when operating near the nanobubble resonance frequency. Here we show that coupled with low energy insonation at a frequency of 80 kHz, well below the resonance frequency of these agents, nanobubbles serve as noninvasive therapeutic warheads that trigger potent mechanical effects in tumors following a systemic injection. We demonstrate these capabilities in tissue mimicking phantoms, where a comparison of the acoustic response of micro- and nano-bubbles after insonation at a frequency of 250 or 80 kHz revealed that higher pressures were needed to implode the nanobubbles compared to microbubbles. Complete nanobubble destruction was achieved at a mechanical index of 2.6 for the 250 kHz insonation vs. 1.2 for the 80 kHz frequency. Thus, the 80 kHz insonation complies with safety regulations that recommend operation below a mechanical index of 1.9. In vitro in breast cancer tumor cells, the cell viability was reduced to 17.3 ± 1.7% of live cells. In vivo, in a breast cancer tumor mouse model, nanobubble tumor distribution and accumulation were evaluated by high frequency ultrasound imaging. Finally, nanobubble-mediated low frequency insonation of breast cancer tumors resulted in effective mechanical tumor ablation and tumor tissue fractionation. This approach provides a unique theranostic platform for safe, noninvasive and low energy tumor mechanotherapy.