Cavitation-induced release of liposomal chemotherapy in orthotopic murine pancreatic cancer models: A feasibility study.

Targeted and triggered release of liposomal drug using ultrasound (US) induced cavitation represents a promising treatment modality to increase the therapeutic-toxicity ratio of encapsulated chemotherapy. OBJECTIVES: To study the feasibility and efficacy of a combination of focused US and liposomal doxorubicin (US-L-DOX) release in orthotopic murine models of pancreatic cancer. MATERIAL AND METHODS: A confocal US setup was developed to generate US inertial cavitation delivery in a controlled and reproducible manner and designed for two distinct murine orthotopic pancreatic cancer models. Controlled cavitation at 1 MHz was applied within the tumors after L-DOX injection according to a preliminary pharmacokinetic study. RESULTS: In vitro studies confirmed that L-DOX was cytostatic. In vivo pharmacokinetic study showed L-DOX peak tumor accumulation at 48h. Feasibility of L-DOX injection and US delivery was demonstrated in both murine models. In a nude mouse model, at W9 after implantation (W5 after treatment), US-L-DOX group (median [IQR] 51.43 mm3 [35.1-871.95]) exhibited significantly lower tumor volumes than the sham group (216.28 [96.12-1202.92]), the US group (359.44 [131.48-1649.25]), and the L-DOX group (255.94 [84.09-943.72]), and a trend, although not statistically significant, to a lower volume than Gemcitabine group (90.48 [42.14-367.78]). CONCLUSION: This study demonstrates that inertial cavitation can be generated to increase the therapeutic effect of drug-carrying liposomes accumulated in the tumor. This approach is potentially an important step towards a therapeutic application of cavitation-induced drug delivery in pancreatic cancer.

Therapeutic efficacy of the combination of doxorubicin-loaded liposomes with inertial cavitation generated by confocal ultrasound in AT2 Dunning rat tumour model.

The combination of liposomal doxorubicin (DXR) and confocal ultrasound (US) was investigated for the enhancement of drug delivery in a rat tumour model. The liposomes, based on the unsaturated phospholipid dierucoylphosphocholine, were designed to be stable during blood circulation in order to maximize accumulation in tumour tissue and to release drug content upon US stimulation. A confocal US setup was developed for delivering inertial cavitation to tumours in a well-controlled and reproducible manner. In vitro studies confirm drug release from liposomes as a function of inertial cavitation dose, while in vivo pharmacokinetic studies show long blood circulation times and peak tumour accumulation at 24-48 h post intravenous administration. Animals injected 6 mg kg(-1) liposomal DXR exposed to US treatment 48 h after administration show significant tumour growth delay compared to control groups. A liposomal DXR dose of 3 mg kg(-1), however, did not induce any significant therapeutic response. This study demonstrates that inertial cavitation can be generated in such a fashion as to disrupt drug carrying liposomes which have accumulated in the tumour, and thereby increase therapeutic effect with a minimum direct effect on the tissue. Such an approach is an important step towards a therapeutic application of cavitation-induced drug delivery and reduced chemotherapy toxicity.

In vivo monitoring of liposomal release in tumours following ultrasound stimulation.

Dioeleoylphosphatidylethanolamine (DOPE)-based liposomes were recently reported as a new class of liposomes for ultrasound (US)-mediated drug delivery. The liposomes showed both high stability and in vitro US-mediated drug release (sonosensitivity). In the current study, in vivo proof-of-principle of US triggered release in tumoured mice was demonstrated using optical imaging. Confocal non-thermal US was used to deliver cavitation to tumours in a well-controlled manner. To detect in vivo release, the near infrared fluorochrome Al (III) Phthalocyanine Chloride Tetrasulphonic acid (AlPcS4) was encapsulated into both DOPE-based liposomes and control liposomes based on hydrogenated soy phosphatidylcholine (HSPC). Encapsulation causes concentration dependent quenching of fluorescence that is recovered upon AlPcS4 release from the liposomes. Exposure of tumours to US resulted in a significant increase in fluorescence in mice administered with DOPE-based liposomes, but no change in the mice treated with HSPC-based liposomes. Thus, DOPE-based liposomes showed superior sonosensitivity compared to HSPC-based liposomes in vivo.

Reflection and transmission at normal incidence onto air-saturated porous materials and direct measurements based on parametric demodulated ultrasonic waves.

The present work is related to the characterization of air-saturated porous media by using parametric demodulated ultrasonic waves. One uses two different powerful ultrasonic emitters working either at 47 kHz or at 162 kHz which are electronically amplitude modulated over the 200 Hz-4 kHz or 2 kHz-40 kHz bandwidths respectively. The demodulation process takes place in air, due to its nonlinearity enabling to generate audio range acoustical waves or alternatively low frequency ultrasonic waves which can be used to characterize porous materials in the reflection configuration at normal incidence. Some appropriate theoretical calculations are introduced for three configurations of interest, i.e. a porous slab, a porous layer mounted onto a rigid plate, and a porous half space, in the case of the equivalent-fluid model. Comparisons between theoretical modeling and experimental data are provided and prospective industrial applications are discussed.

Ultrasonic cavitation in thin liquid layers.

The generation of ultrasonic cavitation in a thin liquid layer trapped between a large radiating surface and a hard reflector and bounded laterally by a gas-liquid interface is investigated. The theoretical analysis predicts that a large amplification of the acoustical pressure is obtained with this configuration. Experiments are conducted by driving the layer with horn-type transducers having a large emitting surface. Ultrasonic cavitation is obtained in a broad frequency range at low input intensity due to the amplification effect. Erosion tests on metallic foils demonstrate the existence of a region of intense cavitation activity which can be localised by controlling the input intensity.

Experimental and theoretical investigation of the mean acoustic pressure in the cavitation field.

The cavitation field radiated by a 20 kHz sonotrode-type transducer is experimentally and theoretically analyzed. Special interest is paid to the origin of the strong fluid streaming appearing in low frequency sonoreactors. A new experimental procedure is proposed to evaluate the mean acoustic pressure inside the fluid. This parameter has been quantified for different points and amplitudes. The velocity of the radiating surface is controlled by a laser interferometer and is always sinusoidal. Train wave excitation is used. The pressure wave and amplitude are measured in the tank with a calibrated hydrophone. The acoustic mean pressure is estimated from the total pressure value at the end of the pulse after an adequate filtering. An analytical nonlinear second order model based on the coupling of the equations of the fluid mechanics with the Rayleigh-Plesset equation is developed in order to relate the measured acoustic parameters to the cavitation state of the fluid. The distributions of the fundamental amplitude and mean pressure are calculated as a function of bubble density and bubble size. A qualitative theoretical description of the experimental data is presented. Quantitative differences and model limitations are commented.

Cone-like bubble formation in ultrasonic cavitation field.

A new phenomenon in ultrasonic cavitation field is reported. Cavitation bubbles are observed to self-arrange in a cone-like macrostructure in the vicinity of transducer radiating surface. The cone-like macrostructure is stable while its branch-like pattern microstructure changes rapidly. The structure is constituted by moving bubbles which undergo attractive and repulsive Bjerknes forces caused by high acoustic pressure gradients and strongly nonlinear oscillations of cavitation bubbles. The cone-like bubble structure is a chemically active formation. Its remarkably high activity is confirmed by chemiluminescence experiments.

Self-induced hysteresis for nonlinear acoustic waves in cracked material.

A new phenomenon of self-induced hysteresis has been observed in the interaction of bulk acoustic waves with a cracked solid. It consists in a hysteretic behavior of material nonlinearity as a function of the incident pump wave amplitude. Hysteresis manifests itself in the self-action of the monochromatic pump wave and in the excitation of its superharmonics and of its subharmonics. The proposed theoretical models attribute the phenomenon to hysteresis in transition of the acoustically forced oscillation of cracks from a nonclapping regime to a regime of clapping contacts.