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.

Water-jet dissector for endoscopic submucosal dissection in an animal study: outcomes of the continuous and pulsed modes.

BACKGROUND: Endoscopic submucosal dissection (ESD) allows en bloc resection of early neoplastic lesions of gastrointestinal tract. Lesions are lifted by submucosal fluid injection before circumferential incision and dissection. High-pressure fluid injection using water jet (WJ) technology is already used for lifting and dissection in surgery. The study was designed to assess WJ for ESD submucosal lifting and dissection. METHODS: An experimental, randomized comparative, "in vivo" nonsurvival animal study on 12 pigs was designed. Stomach mucosal areas were delineated and resected using three ESD techniques: technique A-syringe injection and IT knife dissection; technique B-WJ continuous injection and IT knife dissection; technique C-WJ injection and WJ pulsed dissection. Injection and dissection speeds and complications rates were assessed. RESULTS: Water jet continuous injection is faster than syringe injection (B faster than A, p = 0.001 and B nonsignificantly faster than C, p = 0.06). IT knife dissection is significantly faster after WJ continuous injection (B faster than A, p = 0.003). WJ pulsed dissection is significantly slower than IT knife dissection (C slower than A and B, both p < 0.001). The overall procedure speed was significantly higher and the immediate bleedings rate was significantly lower for technique B than A and C (overall procedure speed p = 0.001, immediate bleedings p = 0.032 and 0.038 respectively). There were no perforations with any technique. CONCLUSIONS: Water jet fluid continuous injection speeds up ESD, whereas pulsed WJ dissection does not.

Feasibility study of cavitation-induced liposomal doxorubicin release in an AT2 Dunning rat tumor model.

BACKGROUND: Targeted and triggered release of liposomal drug using heat or ultrasound represents a promising treatment modality able to increase the therapeutic-totoxicity ratio of encapsulated drugs. PURPOSE: To study the ability for high-intensity focused ultrasound to induce liposomal drug release mainly by focused inertial cavitation in vitro and in an animal model. METHODS: A 1 MHz ultrasound setup has been developed for in vitro and in vivo drug release from a specific liposomal doxorubicin formulation at a target cavitation dose. RESULTS: Controlled cavitation at 1 MHz was applied within the tumors 48 hours after liposome injection according to preliminary pharmacokinetic study. A small non-significant therapeutic effect of US-liposomal treatment was observed compared to liposomes alone suggesting no beneficial effect of ultrasound in the current setup. CONCLUSION: The in vitro study provided a suitable ultrasound setup for delivering a cavitation dose appropriate for safe liposomal drug release. However, when converting to an in vivo model, no therapeutic benefit was observed. This may be due to a number of reasons, one of which may be the difficulty in converting in vitro findings to an in vivo model. In light of these findings, we discuss important design features for future studies.

Thermal ablation by high-intensity-focused ultrasound using a toroid transducer increases the coagulated volume. Results of animal experiments.

Surgical resection is the only treatment of colorectal liver metastases that can ensure long-term survival and cure in some patients. However, only 20% of patients are suitable for surgery. As a result, many nonresectional modalities of treatment have been assessed to provide an alternative to liver resection. Several limitations have been observed when using these techniques and available evidence is limited. Here, we report that a new design of high intensity focused ultrasound transducer can significantly enlarge the coagulated volume over short periods of time and that treatment in the liver can be guided in real-time using an integrated ultrasound imaging probe. Our long-term objective is to develop a device that can be used during surgery for eventual clinical use in conjunction with resection. Eight ultrasound emitters, divided into 256 elements, were created by sectioning a single toroid piezocomposite transducer. The focal zone was conical in shape and located 70 mm from the transducer; enabling the treatment of deep-seated tumors. A single thermal lesion was created when the eight emitters performed alternative and consecutive 5-s ultrasound exposures. This article presents in vivo evidence that the coagulated volume obtained from a 40 s total exposure in the liver was 7.0 +/- 2.5 cm(3) (minimum 1.5 - maximum 20.0 cm(3)) with an average diameter of 17.5 +/- 3.8 mm (minimum 10.0 - maximum 29.0 mm). All lesions were visible with high contrast on sonograms. The correlation between the diameter of lesions observed on sonograms and during gross examination was 92%. This method also allowed the user to easily enlarge the coagulated volume by juxtaposing single lesions. This approach may have a role in treating unresectable colorectal liver metastases and may also be used in conjunction with resection to extend its limits.

High-intensity focused ultrasound ablation for the treatment of colorectal liver metastases during an open procedure: study on the pig.

OBJECTIVE: To demonstrate in a porcine model that high-intensity focused ultrasound (HIFU) with toroid-shaped emitters may have a role in treating unresectable colorectal liver metastases. SUMMARY BACKGROUND DATA: Surgical resection is the only curative option for colorectal hepatic metastases. Only 20% of patients are suitable for surgery. Many ablative techniques have been assessed but several limitations have been documented: traumatic puncture of the parenchyma, limited size of lesions, and inability to monitor the treatment in real time. METHODS: A HIFU device with 256 toroid-shaped emitters and integrated ultrasound imaging probe was used. Single lesions, induced in 40 seconds, and juxtaposition of 6 single lesions were created under ultrasound guidance on 13 pigs. The lesions were studied on sonograms, macroscopically and microscopically up to 30 days after the treatment. RESULTS: Ninety percent of the HIFU lesions were immediately hypoechoic on ultrasound imaging. The average coagulated volume obtained from a 40 seconds total exposure in the liver was 7.0 +/- 2.5 cm (1.5-20.0), average diameter: 19.5 +/- 3.8 mm (10.0-29.0). Using the real-time visualization of the treated region, single lesions were easily juxtaposed to produce larger lesions up to 6 cm in diameter without any major complication. CONCLUSIONS: This toroid HIFU device allows short treatment times, noninvasiveness regarding the liver and real time ultrasound guidance. It seems to be simpler and more reliable to use than current ablative methods. Additionally, lesions through large vessels (up to 5 mm) being feasible, treatment of some juxta-vascular metastases should be possible.

Percutaneous sonographically guided interstitial US ablation: experimentation in an in vivo pig liver model.

PURPOSE: To test the feasibility and efficacy of a percutaneous sonographically guided high-intensity interstitial ultrasound (US) ablation applicator to create "macrolesions" of confluent coagulation in an in vivo pig liver model. MATERIALS AND METHODS: Eight pigs and an interstitial US ablation applicator were used for this study. Elementary lesions and macrolesions created by the confluence of several elementary lesions were successively analyzed. The first phase of the study was performed by varying the acoustic intensity (AI) to create elementary lesions. The second phase of the study aimed at creating macrolesions in three target zones previously defined in the liver. Mean (+/- SD) maximum and minimum diameters of the macrolesions were calculated. RESULTS: The elementary lesions created with AIs of 30, 40, and 55 W/cm(2) measured 14.9 mm +/- 3, 19.8 mm +/- 5, and 13.2 mm +/- 3, respectively. The AI retained for the second experimental phase was 40 W/cm(2). Because of the flamelike aspect of the elementary lesions, the macrolesion appeared as a cylinder with crenelated contours. On macroscopic examination, mean maximum diameters of the macrolesions were 28.7 mm +/- 3, 34.1 mm +/- 2, and 27.8 mm +/- 5 and mean minimum diameters of the macrolesions were 14 mm +/- 3, 18.7 mm +/- 2, and 14 mm +/- 3 for the three target zones, respectively. A single major complication, puncture of the gallbladder, occurred in one animal. CONCLUSIONS: A percutaneous sonographically guided interstitial ablation applicator is able to create macrolesions of coagulation in pig liver.

Utility of a tumor-mimic model for the evaluation of the accuracy of HIFU treatments. results of in vitro experiments in the liver.

Presented in this article is a tumor-mimic model that allows the evaluation, before clinical trials, of the targeting accuracy of a high intensity focused ultrasound (HIFU) device for the treatment of the liver. The tumor-mimic models are made by injecting a warm solution that polymerizes in hepatic tissue and forms a 1 cm discrete lesion that is detectable by ultrasound imaging and gross pathology. First, the acoustical characteristics of the tumor-mimics model were measured in order to determine if this model could be used as a target for the evaluation of the accuracy of HIFU treatments without modifying HIFU lesions in terms of size, shape and homogeneity. On average (n = 10), the attenuation was 0.39 +/- 0.05 dB.cm(-1) at 1 MHz, the ultrasound propagation velocity was 1523 +/- 1 m.s(-1) and the acoustic impedance was 1.84 +/- 0.00 MRayls. Next, the tumor-mimic models were used in vitro in order to verify, at a preclinical stage, that lesions created by HIFU devices guided by ultrasound imaging are properly positioned in tissues. The HIFU device used in this study is a 256-element phased-array toroid transducer working at a frequency of 3 MHz with an integrated ultrasound imaging probe working at a frequency of 7.5 MHz. An initial series of in vitro experiments has shown that there is no significant difference in the dimensions of the HIFU lesions created in the liver with or without tumor-mimic models (p = 0.3049 and p = 0.8796 for the diameter and depth, respectively). A second in vitro study showed that HIFU treatments performed on five tumor-mimics with safety margins of at least 1 mm were properly positioned. The margins obtained were on average 9.3 +/- 2.7 mm (min. 3.0 - max. 20.0 mm). This article presents in vitro evidence that these tumor-mimics are identifiable by ultrasound imaging, they do not modify the geometry of HIFU lesions and, thus, they constitute a viable model of tumor-mimics indicated for HIFU therapy.

Delivery by shock waves of active principle embedded in gelatin-based capsules.

PURPOSE: Delivering a drug close to the targeted cells improves its benefit versus risk ratio. A possible method for local drug delivery is to encapsulate the drug into solid microscopic carriers and to release it by ultrasound. The objective of this work was to use shock waves for delivering a molecule loaded in polymeric microcapsules. MATERIAL AND METHODS: Ethyl benzoate (EBZ) was encapsulated in spherical gelatin shells by complex coacervation. A piezocomposite shock wave generator (120 mm in diameter, focused at 97 mm, pulse length 1.4 micros) was used for sonicating the capsules and delivering the molecule. Shock parameters (acoustic pressure, number of shocks and shock repetition frequency) were varied in order to measure their influence on EBZ release. A cavitation-inhibitor liquid (Ablasonic) was then used to evaluate the role of cavitation in the capsule disruption. RESULTS: The measurements showed that the mean quantity of released EBZ was proportional to the acoustic pressure of the shock wave (r2 > 0.99), and increased with the number of applied shocks. Up to 88% of encapsulated EBZ could be released within 4 min only (240 shocks, 1 Hz). However, the quantity of released EBZ dropped at high shock rates (above 2Hz). Ultrasound imaging sequences showed that cavitation clouds might form, at high shock rates, along the acoustic axis making the exposure inefficient. Measurements done in Ablasonic showed that cavitation plays a major role in microcapsules disruption. CONCLUSIONS: In this study, we designed polymeric capsules that can be disrupted by shock waves. This type of microcapsule is theoretically a suitable vehicle for carrying hydrophobic drugs. Following these positive results, encapsulation of drugs is considered for further medical applications.

Synergistic inhibitory effect of high-intensity focused ultrasound combined with chemotherapy on Dunning adenocarcinoma.

OBJECTIVE: To evaluate the therapeutic effect of high-intensity focused ultrasound (HIFU) combined with chemotherapy (paclitaxel + estramustine) on AT2 Dunning adenocarcinoma, as no satisfactory treatment for localized prostate cancer is available for patients with a poor prognosis, e.g. stage T3, a high Gleason score, or a prostate-specific antigen level of >15 ng/mL. MATERIALS AND METHODS: Forty-one Dunning AT2 tumour-bearing Copenhagen rats were divided into four groups, i.e. control, chemotherapy, HIFU, and chemotherapy + HIFU (the last three treated for 1 week). The growth in tumour volume was recorded for 3 weeks, the point at which tumour volume was considered to have doubled (doubling time). The growth curves of each group were plotted and evaluated statistically. RESULTS: At 30 days of follow-up the distributions of tumour volume with treatment group were significantly different (P < 0.001); volumes were significantly greater in the control than in the chemotherapy-only or in the HIFU-only group (both P = 0.006). The greatest difference was between the chemotherapy + HIFU and the control group. The tumour doubling times were 13.2 days for HIFU-only, 31.2 days for chemotherapy + HIFU and 7.7 days for the controls. CONCLUSION: These results suggest that this combined therapy could be useful for treating patients with high-risk prostate cancer.

Impedance changes in liver tissue exposed in vitro to high-energy ultrasound.

The present study reports the impedance changes observed in bovine liver samples exposed in vitro to high-intensity ultrasound. The measurement frequency ranged from 80 kHz to 2 MHz. The treatment resulted in the average increase of 20% in impedance magnitude at low frequency and the average decrease of 30% at high frequency. The phase angle increased significantly by more than 15 degrees at all measurement frequencies. The slope of the log-modulus of impedance against log-frequency increased in treated tissue at frequencies above 500 kHz. This change was attributed to the alteration of the capacitive response of the tissue. The experimental observations are consistent with the known changes induced by high-energy ultrasound in liver tissue. This study confirmed that ultrasound energy produces measurable changes in a tissue's impedance and that indices can be derived to distinguish between original and treated tissues. The results obtained in liver tissue need confirmation in organs treatable with therapeutic ultrasound, such as breast and prostate.

[Potentiation of focal ultrasound treatment of prostate adenocarcinoma by concomitant chemotherapy with estramustine phosphate and paclitaxel]. / Potentialisation du traitement par ultrasons focalisés de l'adénocarcinome prostatique par une chimiothérapie concomitante associant estramustine phosphate et paclitaxel.

STUDY OBJECTIVES: To demonstrate a synergistic action between high intensity focal ultrasound (HIFU) and combination chemotherapy with paclitaxel and estramustine phosphate (EMP) on a prostate cancer model. MATERIAL AND METHODS: The animal model used in this study was the Copenhagen rat and the tumour model is a Dunning R 3327-AT 2 hormone-independent prostatic adenocarcinoma cell line. Chemotherapy was administered once a week for 4 weeks according to 2 modalities: low-dose with paclitaxel 2 mg/kg/day, 1 day per week and EMP 50 mg/kg/day, 3 days per week; or high-dose with paclitaxel 3 mg/kg/day, 1 day per week and EMP 75 mg/kg/day, 3 days per week. Treatment with HIFU was performed at the second week and only 55% of the tumour volume was treated. The study was conducted on 42 rats divided into 6 arms: Control, HIFU, low-dose paclitaxel-EMP, high-dose paclitaxel-EMP, HIFU + low-dose paclitaxel-EMP and HIFU + high-dose paclitaxel-EMP. Study endpoints were the course of tumour volume and animal survival. RESULTS: After two weeks of treatment, a statistically significant difference for tumour volume was observed between the various arms of the study (p < 0.0001). The HIFU-chemotherapy arm and, to a lesser degree, the chemotherapy only arm, presented the lowest tumour progression. CONCLUSION: The combination of HIFU + paclitaxel-EMP is more effective than treatment with HIFU alone or paclitaxel-EMP alone on growth of the Dunning tumour, right from the first weeks of treatment.

Feasibility of a transurethral ultrasound applicator for coagulation in prostate.

Transurethral resection of the prostate (TURP) is the surgical method routinely used in clinics to treat benign prostate hyperplasia (BPH). The purpose of this work is to demonstrate the feasibility of a transurethral ultrasound (US) applicator based on a miniature US flat transducer to coagulate prostatic tissues. Rabbit liver was found to comply well with human prostate. A significant fall in Doppler signal amplitude immediately after treatment demonstrated the applicator's ability to achieve haemostasis. The therapeutic depth extended from 6 to 10 mm, depending on conditions of exposure, and the coagulation rate ranged between 51% and 99%. The coagulated zone pinpointed on histological examination could be easily correlated to a permanent hypoechoic zone observed on B-scans of treated zones. This observation is most likely due to temperature-related changes in the acoustic attenuation of liver and, unfortunately, may not be visible in the prostate.