Ultrasound-Mediated Blood-Brain Barrier Opening Improves Whole Brain Gene Delivery in Mice.

Gene therapy represents a powerful therapeutic tool to treat diseased tissues and provide a durable and effective correction. The central nervous system (CNS) is the target of many gene therapy protocols, but its high complexity makes it one of the most difficult organs to reach, in part due to the blood-brain barrier that protects it from external threats. Focused ultrasound (FUS) coupled with microbubbles appears as a technological breakthrough to deliver therapeutic agents into the CNS. While most studies focus on a specific targeted area of the brain, the present work proposes to permeabilize the entire brain for gene therapy in several pathologies. Our results show that, after i.v. administration and FUS sonication in a raster scan manner, a self-complementary AAV9-CMV-GFP vector strongly and safely infected the whole brain of mice. An increase in vector DNA (19.8 times), GFP mRNA (16.4 times), and GFP protein levels (17.4 times) was measured in whole brain extracts of FUS-treated GFP injected mice compared to non-FUS GFP injected mice. In addition to this increase in GFP levels, on average, a 7.3-fold increase of infected cells in the cortex, hippocampus, and striatum was observed. No side effects were detected in the brain of treated mice. The combining of FUS and AAV-based gene delivery represents a significant improvement in the treatment of neurological genetic diseases.

Development of a multi-functional preclinical device for the treatment of glioblastoma.

Glioblastoma multiforme (GBM) is one of the most common and aggressive malignant primary brain tumors in adults. The treatment of GBM is limited by the blood-brain barrier (BBB), which limits the diffusion of appropriate concentrations of therapeutic agents at the tumor site. Among experimental therapies, photo-thermal therapy (PTT) mediated by nanoparticles is a promising strategy. To propose a preclinical versatile research instrument for the development of new PTT for GBM, a multipurpose integrated preclinical device was developed. The setup is able to perform: i) BBB permeabilization by focused ultrasound sonication (FUS); ii) PTT with continuous wave laser; iii) in situ temperature monitoring with photo-acoustic (PA) measurements. In vivo preliminary subcutaneous and transcranial experiments were conducted on healthy or tumor-bearing mice. Transcranial FUS-induced BBB permeabilization was validated using single photon emission computed tomography (SPECT) imaging. PTT capacities were monitored by PA thermometry, and are illustrated through subcutaneous and transcranial in vivo experiments. The results show the therapeutic possibilities and ergonomy of such integrated device as a tool for the validation of future treatments.

Laser Ablation-Assisted Synthesis of Plasmonic Si@Au Core-Satellite Nanocomposites for Biomedical Applications.

Owing to strong plasmonic absorption and excellent biocompatibility, gold nanostructures are among best candidates for photoacoustic bioimaging and photothermal therapy, but such applications require ultrapure Au-based nanoformulations of complex geometry (core-shells, nanorods) in order to shift the absorption band toward the region of relative tissue transparency (650-1000 nm). Here, we present a methodology for the fabrication of Si@Au core-satellite nanostructures, comprising of a Si core covered with small Au nanoparticles (NP), based on laser ablative synthesis of Si and Au NPs in water/ethanol solutions, followed by a chemical modification of the Si NPs by 3-aminopropyltrimethoxysilane (APTMS) and their subsequent decoration by the Au NPs. We show that the formed core-satellites have a red-shifted plasmonic absorption feature compared to that of pure Au NPs (520 nm), with the position of the peak depending on APTMS amount, water-ethanol solvent percentage and Si-Au volume ratio. As an example, even relatively small 40-nm core-satellites (34 nm Si core + 4 nm Au shell) provided a much red shifted peak centered around 610 nm and having a large tail over 700 nm. The generation of the plasmonic peak is confirmed by modeling of Si@Au core-shells of relevant parameters via Mie theory. Being relatively small and exempt of any toxic impurity due to ultraclean laser synthesis, the Si@Au core-satellites promise a major advancement of imaging and phototherapy modalities based on plasmonic properties of nanomaterials.

Acoustical characterisation and monitoring of microbubble clouds.

Argon microbubbles will exist in the primary sodium of the next generation of sodium-cooled fast reactors (SFR). Due to its opacity, acoustic methods will be used for the in-service inspection in these reactors, but the presence of such bubbles will greatly affect ultrasonic wave propagation. Moreover, these bubbles can lead to the formation of gas pockets in the reactor and impact cavitation and boiling phenomena. It is therefore necessary to characterise what is called the 'microbubble cloud' by providing the volume fraction and the bubble size distribution. Safety requirements in this field call for robust inspection methods based on very few assumptions about the bubble populations. The objective of this study is to assess the performance of spectroscopic methods in the presence of bubbles with high polydispersity and to monitor an evolving cloud of microbubbles. The histogram and void fractions were estimated according to the regularised inversion of the complex wave number's integral equation. To reduce the need for prior information on the bubble cloud, a specific procedure was used to estimate the maximum radius of the population. The results are presented on the basis of the experimental data obtained and then compared with optical measurements.

Bubbly flow velocity measurement in multiple scattering regime.

We propose a technique to measure the velocity of a bubble cloud based on the coda correlation. The method is founded on successive recordings of multiple scattered waves from a bubble cloud. Our model predicts the dependence between the correlation coefficient of these coda waves and the velocity of the bubble cloud under diffusion approximation. The Acoustic experiments are validated by simultaneous optical measurements in a water tank, with a good agreement between the acoustical and the optical methods (relative difference smaller than 7%). This technique can be transposed to any particle flow velocity problems involving multiple scattering effects in acoustics.

Laser- synthesized TiN nanoparticles as promising plasmonic alternative for biomedical applications.

Exhibiting a red-shifted absorption/scattering feature compared to conventional plasmonic metals, titanium nitride nanoparticles (TiN NPs) look as very promising candidates for biomedical applications, but these applications are still underexplored despite the presence of extensive data for conventional plasmonic counterparts. Here, we report the fabrication of ultrapure, size-tunable TiN NPs by methods of femtosecond laser ablation in liquids and their biological testing. We show that TiN NPs demonstrate strong and broad plasmonic peak around 640-700 nm with a significant tail over 800 nm even for small NPs sizes (<7 nm). In vitro tests of laser-synthesized TiN NPs on cellular models evidence their low cytotoxicity and excellent cell uptake. We finally demonstrate a strong photothermal therapy effect on U87-MG cancer cell cultures using TiN NPs as sensitizers of local hyperthermia under near-infrared laser excitation. Based on absorption band in the region of relative tissue transparency and acceptable biocompatibility, laser-synthesized TiN NPs promise the advancement of biomedical modalities employing plasmonic effects, including absorption/scattering contrast imaging, photothermal therapy, photoacoustic imaging and SERS.

Weld inspection by focused adjoint method.

This paper reports a methodology for the non-destructive ultrasonic evaluation of welds, based on probing, residue back-focusing and topological energy calculation using an enhanced (focused) adjoint method. The proposed method combines the advantages of time reversal to compensate for the cumulative distorsions experienced by a wave propagating in a heterogeneous medium, and topological imaging to highlight the defect location. The synergistic effect of this combined approach makes it possible to detect anomalies in the most efficient way. The method paves the way towards a matched-insonification imaging of anomalies in anisotropic media.

Topological imaging in bounded elastic media.

Detecting, imaging and sizing defects in a bounded elastic medium is still a difficult task, especially when access is complex. Adjoint methods simplify the task as they take advantage of prior information such as the geometry and material properties. However, they still reveal a number of important limitations. Artifacts observed on the conventional topological energy image result from wave interactions with the boundaries of the inspected medium. The paper describes a method for addressing these artifacts, which involves forward and adjoint fields specified in terms of the boundary conditions. Modified topological energies are then defined according to the type of analyzed flaw (open slit or inclusion). Comparison of the numerical results with the experimental data confirms the relevance of the approach.

Taking advantage of acoustic inhomogeneities in photoacoustic measurements.

This paper proposes a method for improving the localization and the quantification of the optical parameters in photoacoustic (PA) tomography of biological tissues that are intrinsically heterogeneous in both optical and acoustic properties. It is based on the exploitation of both the PA signal, generated by the heterogeneous optical structures, and the secondary acoustic echoes due to the interaction between a primary PA wave generated near the tissue surface and the heterogeneous acoustic structures. These secondary echoes can also be collected through proper measurements of the PA signals. The experimental procedure is presented along with the method to filter the signal and the reconstruction algorithm that includes the account of the acoustic information.

Microbubble dynamics monitoring using a dual modulation method.

An experimental method for characterizing microbubbles' oscillations is presented. With a Dual Frequency ultrasound excitation method, both relative and absolute microbubble size variations can be measured. Using the same experimental setup, a simple signal processing step applied to both the amplitude and the frequency modulations yields a two-fold picture of microbubbles' dynamics. In addition, assuming the occurrence of small radial oscillations, the equilibrium radius of the microbubbles can be accurately estimated.

The benefits of compression methods in acoustic coherence tomography.

Pulse compression methods improve the quality of ultrasonic medical images. In comparison with standard broadband pulse techniques, these methods enhance the contrast-to-noise ratio (CNR) and increase the probing depth without any perceptible loss of spatial resolution. The Golay compression technique is analyzed here in the context of ultrasonic computed tomography, first on a one-dimensional target and second on a very low-contrast phantom probed using a half-ring array tomograph. The imaging performances were assessed based on the image CNR. The improvement obtained (up to 40%) depends, however, on the number of coherently associated diffraction projections. Beyond a certain number, few advantages were observed. Advances in ultrasound computed tomography suggest that pulse compression methods should provide a useful means of optimizing the trade-off between the image quality and the probing sampling density.

Conformal ultrasound imaging system for anatomical breast inspection.

Ultrasound tomography has considerable potential as a means of breast cancer detection because it reduces the operator-dependency observed in echography. A half-ring transducer array was designed based on breast anatomy, to obtain reflectivity images of the ductolobular structures using tomographic reconstruction procedures. The 3-MHz transducer array comprises 1024 elements set in a 190-degree circular arc with a radius of 100 mm. The front-end electronics incorporate 32 independent parallel transmit/receive channels and a 32-to-1024 multiplexer unit. The transmit and receive circuitries have a variable sampling frequency of up to 80 MHz and 12-bit precision. Arbitrary waveforms are synthesized to improve the signal-to-noise ratio and to increase the spatial resolution when working with low-contrast objects. The setup was calibrated with academic objects and a needle hydrophone to develop the data correction tools and specify the properties of the system. The backscattering field was recorded using a restricted aperture, and tomographic acquisitions were performed with a pair of 0.08-mm-diameter steel wires, a low-contrast 2-D breast phantom, and a breast-shaped phantom containing inclusions. Data were processed with dedicated correction tools and a pulse compression technique. Objects were reconstructed using the elliptical back-projection algorithm.

An optimization method for quantitative impedance tomography.

A near-field ultrasonic tomography method providing high resolution imaging for soft tissue in the reflection mode is reported. When the Born approximation is valid, the main limitation of this method is that it requires an incident pulse with infinite bandwidth, whereas the incident pulses used in practice have a limited bandwidth, which makes quantitative reconstruction impossible. The reconstructed image is qualitative in the sense that it is a band-pass filtered reconstruction of the impedance distribution. An optimization method based on the use of the geometrical information provided by the tomographic reconstruction is developed to obtain the quantitative information required. The object was approximated locally by an equivalent canonical body, on the basis of the previous global estimation. The inversion procedure is then carried out using the minimization of a cost function, which is the average over frequency of the difference between the measured field scattered by the object and the estimated field scattered by the equivalent canonical body. Assuming the object to be homogeneous by regions, the last step consists of assigning the estimated local impedance value to the region of interest. When the geometry of the real body is almost canonical, the optimization method yields accurate impedance assessments.

Near-field ultrasound tomography.

In this paper, a near-field tomographic solution is introduced to solve the imaging problem of fluid objects assumed to be weakly heterogeneous (Born approximation) and excited by spherical waves. The solution to the forward problem is based on the Huygens-Fresnel principle which describes the scattered field as the result of the interference scheme of all the secondary spherical waves. From the derivation of the scattered field, a new Fourier transform that has been called the elliptical Fourier transform is defined: It differs from the standard Fourier transform in that instead of a plane wave decomposition, a harmonic ellipsoidal wave decomposition is obtained. Based on this spectral analysis, a near-field Radon transform is designed that complements the "far-field tools" published in diffraction tomography literature. Then, assuming that the measuring distance is greater than one wavelength, the feasibility of reconstructing either the impedance or the velocity maps of an acoustical (perfect fluid) model is demonstrated. Numerical simulations were performed which confirmed the validity of the theory presented here; a theory which has many potential applications in future wave theory research.

A 2-D anatomic breast ductal computer phantom for ultrasonic imaging.

Most breast cancers (85%) originate from the epithelium and develop first in the ductolobular structures. In screening procedures, the mammary epithelium should therefore be investigated first by the performing of an anatomically guided examination. For this purpose (mass screening, surgical guidance), we developed a two-dimensional anatomic phantom corresponding to an axial cross section of the ductolobular structures, which makes it possible to better understand the interactions between the breast composition and ultrasound. The various constitutive tissues were modeled as a random inhomogeneous continuum with density and sound speed fluctuations. Ultrasonic pulse propagation through the breast computer phantom was simulated using a finite element time domain method (the phantom can be used with other propagation codes). The simulated ductal echographic image is compared with the ductal tomographic (DT) reconstruction. The preliminary results obtained show that the DT method is more satisfactory in terms of both the contrast and the resolution.

Soft tissue absorption tomography with correction for scattering aberrations.

Among the many factors involved in ultrasound attenuation phenomena, scattering effects play a major role, even in the unexpected case of soft tissues. It is proposed in this study to quantitatively evaluate the scattering affecting the measurements before reconstructing the absorption parameter alone. The reconstruction procedure involves three steps: (1) Estimating the sound speed map using a transmission tomography algorithm. This estimation procedure provides a numerical phantom of the organ probed, cleared of all dissipative components. This absorption free phantom mimics the (viscoacoustic) tissues imaged except for the density and absorption characteristics: the density a priori equals 1000 kg/m3 and the absorption is not taken into account. The impedance fluctuations in the object are therefore approximated on the basis of the sound speed contrast. (2) Synthesing the field scattered by the absorption free phantom; the attenuation observed here results solely from the scattering phenomenon. The synthesis is carried out using a finite-element time domain code simulating the ultrasonic propagation through the phantom. It provides the scattering distortion reference introduced into the log spectral absorption estimator. (3) Reducing the scattering distortions affecting the integrated absorption measured along the ray paths using a log spectral procedure. The corrected integrated absorption is then processed using a tomographic reconstruction procedure that provides an estimate of the absorption distribution. Simple numerical simulations show the improvement obtained in the absorption estimates with this approach.

Weakly inhomogeneous media tomography.

Our objective is to develop an ultrasonic scanner for breast imaging. High resolution is obtained by using wide-band spherical waves transmitted and measured in the near field zone (i.e., close to the skin) all around the organ. The tomographic approach that we adopt allows us to use low central frequency waves (3-7 MHz) that are suitable for good penetration while maintaining high resolution and contrast. The procedure is thus suitable for early detection of tumors and increases the chances of total recovery. The novelty of the present reconstruction procedure is that it associates the signals acquired in transmission to the data measured in reflection over a large aperture. This enables us to correct the phase aberration induced by weak inhomogeneities whose sizes might be several wavelengths. Numerical tests based on Finite Difference Time Domain (FDTD) simulations demonstrate the greater fidelity of the reconstruction.

Near-field diffraction tomography.

We propose an algorithm that deals with broadband ultrasonic signals acquired in the near-field domain using probes close to the skin. This technique is designed for diffraction tomography of compact support objects interrogated by spherical waves (small transducers). It is an approximate inversion procedure in the Born approximation based on elliptical backprojection. Near-field imaging is enhanced by reducing the geometrical distortion observed on standard tomography. Numerical tests based on finite difference time domain (FDTD) simulations of data scattered by a tissue-like phantom are given.