Focused ultrasound on the substantia nigra enables safe neurotensin-polyplex nanoparticle-mediated gene delivery to dopaminergic neurons intranasally and by blood circulation.

Neurotensin-polyplex nanoparticles provide efficient gene transfection of nigral dopaminergic neurons when intracerebrally injected in preclinical trials of Parkinson's disease because they do not cross the blood-brain barrier (BBB). Therefore, this study aimed to open BBB with focused ultrasound (FUS) on the substantia nigra to attain systemic and intranasal transfections and evaluate its detrimental effect in rats. Systemically injected Evans Blue showed that a two-pulse FUS opened the nigral BBB. Accordingly, 35 µL of neurotensin-polyplex nanoparticles encompassing the green fluorescent protein plasmid (79.6 nm mean size and + 1.3 mV Zeta-potential) caused its expression in tyrosine hydroxylase(+) cells (dopaminergic neurons) of both substantiae nigrae upon delivery via internal carotid artery, retro-orbital venous sinus, or nasal mucosa 30 min after FUS. The intracarotid delivery yielded the highest transgene expression, followed by intranasal and venous administration. However, FUS caused neuroinflammation displayed by infiltrated lymphocytes (positive to cluster of differentiation 45), activated microglia (positive to ionized calcium-binding adaptor molecule 1), neurotoxic A1 astrocytes (positive to glial fibrillary acidic protein and complement component 3), and neurotrophic A2 astrocytes (positive to glial fibrillary acidic protein and S100 calcium-binding protein A10), that ended 15 days after FUS. Dopaminergic neurons and axonal projections decreased but recuperated basal values on day 15 after transfection, correlating with a decrease and recovery of locomotor behavior. In conclusion, FUS caused transient neuroinflammation and reversible neuronal affection but allowed systemic and intranasal transfection of dopaminergic neurons in both substantiae nigrae. Therefore, FUS could advance neurotensin-polyplex nanotechnology to clinical trials for Parkinson's disease.

Protocol to Induce the Temporary Opening of the Blood-Brain Barrier with Short-Time Focused Ultrasound in Rats.

Brain neurodegenerative diseases are central nervous system (CNS) affections typically common in older adults. A new therapeutic approach for them consists of providing specific drugs to the CNS through blood circulation; however, the Blood-Brain Barrier (BBB) prevents almost 100% of neurotherapeutics from reaching the brain. There are indications that Focused Ultrasound (FUS), temporarily placed in the BBB, can achieve a controlled increase in temperature at its focus, allowing temporary, localized, and reversible opening of this barrier, which facilitates the temporary delivery of specific drugs. This work presents a FUS-based protocol for the local, temporary, and reversible opening of the BBB in Wistar rats. The proposed protocol specifies certain power, treatment times, and duty cycle to controllably increase the temperature at the region of interest, i.e., the substantia nigra. Numerical simulations using commercial software based on the finite element method were carried out to determine the optimal size of the craniotomies for nearly full-acoustic transmission. Experiments in rats were performed with the parameters used during computational simulations to determine the adequate opening of the BBB. For this, craniotomies of different sizes were made at coordinates of the substantia nigra, and FUS was applied from the exterior. The opening of the BBB was evaluated using Evans Blue (EB) as an indicator of the crossing of the dye from the blood vessels to brain tissue. Numerical simulations demonstrated a major distance reached by the ultrasound focus with a bigger diameter. Experimental results show the local, temporary, and reversible opening of the BBB through a 10 mm diameter craniotomy, which effectively allowed placing the ultrasound focus over the substantia nigra, unlike a 6 mm diameter craniotomy in which there is a deviation of the focus through that window. Moreover, from these results, it was also determined that the disruption of the BBB was reversible, with an opening duration of 6 h after FUS application. The experimental work developed in this study resulted in a minimally invasive method for the temporary opening of the BBB.

Novel Cranial Implants of Yttria-Stabilized Zirconia as Acoustic Windows for Ultrasonic Brain Therapy.

Therapeutic ultrasound can induce changes in tissues by means of thermal and nonthermal effects. It is proposed for treatment of some brain pathologies such as Alzheimer's, Parkinson's, Huntington's diseases, and cancer. However, cranium highly absorbs ultrasound reducing transmission efficiency. There are clinical applications of transcranial focused ultrasound and implantable ultrasound transducers proposed to address this problem. In this paper, biocompatible materials are proposed for replacing part of the cranium (cranial implants) based on low porosity polycrystalline 8 mol% yttria-stabilized-zirconia (8YSZ) ceramics as acoustic windows for brain therapy. In order to assess the viability of 8YSZ implants to effectively transmit ultrasound, various 8YSZ ceramics with different porosity are tested; their acoustic properties are measured; and the results are validated using finite element models simulating wave propagation to brain tissue through 8YSZ windows. The ultrasound attenuation is found to be linearly dependent on ceramics' porosity. Results for the nearly pore-free case indicate that 8YSZ is highly effective in transmitting ultrasound, with overall maximum transmission efficiency of ≈81%, compared to near total absorption of cranial bone. These results suggest that 8YSZ polycrystals could be suitable acoustic windows for ultrasound brain therapy at 1 MHz.

Performance analysis of ultrasono-therapy transducer with contact detection.

The performance of ultrasono-therapy transducer with contact detection by using the impedance phase change is described. Usually a therapy transducer is designed with a lambda/2 frontal plate glued to a PZT-4 piezoceramic. This plate ensures a good mechanical protection of the piezoceramic with a corresponding high-transmission energy. Normally this transducer is operated at the minimum at the frequency of the impedance module of its input electric impedance, but this operation point is affected by the shift caused by the expected temperature increase. This shift could be higher than the narrow bandwidth presented. As a result we obtain a decrease in the power level for medical treatment. Usually it is designed electronic drivers with automatic control that follow the frequency change, but the relatively narrow bandwidth introduces difficulty in the design. Another frequency operation point is presented here and analyzed using the criteria of the maximum of the impedance phase with a wider bandwidth than in the previous case. Simulation with mechanical losses are presented with experimental results that show the convenience of this criteria for practical application.

Acoustic field modeling for physiotherapy ultrasound applicators by using approximated functions of measured non-uniform radiation distributions.

The strongest therapeutic effects in ultrasonic physiotherapy are mainly produced at the first centimeters, i.e. close to the applicator surface and, in general, only in the near-field zone. The acoustic field produced in practice by this type of transducers differs from the classical models because the vibration distribution on the real transducer surfaces is non-uniform. However, neither models using uniform distribution, nor those using typical non-uniform distribution patterns for the source accurately represent the radiation of this kind of transducers. Although this therapy is widely used and many efforts have been made in experimentally studying the patterns of ultrasound radiation produced during physiotherapy applications (IEC-61689, 1998), additional modeling researches still would be needed in order to achieve improved models giving field patterns closer to the measured ultrasonic results. In this paper, acoustic patterns produced from two source radiation functions are proposed and evaluated for field modeling of physiotherapy applicators. Both the functions are approximations to the pressure distribution measured close to the emitting surface and they are based on the modulation of the classical simply-supported function using either sinusoidal or Bessel-type distributions. The simply-supported function is accounted for the radiator-fixing condition and the modulation function simulates the complex vibration distribution of this kind of transducer. The modulator Bessel function is based on reports about Bessel-type vibration distributions found in piezoelectric disk resonators. The use of a selected sinusoidal segment represents another analytical option for obtaining an approximated behavior of the measured data in a real applicator. Both the field models are implemented using the finite element method (FEM) to obtain the numerical solution of wave equation at each point in the radiated space. The solution is reached by considering axisymmetric radiation in attenuation-free media. The results indicate the viability of applying an adequate model for acoustic field calculation by simulating the radiating distribution on the emitting surface as either sinusoidal or Bessel-modulated functions. Models using both the functions describe reasonably real behaviors, but those based on Bessel functions are better correlated with the measurements. The results for three commercial applicators indicate the possibility of representing, with adequate verisimilitude, the acoustic field radiated by physiotherapy ultrasound transducers using linear combinations of Bessel profiles describing the radiation source.

Active upper limb prosthesis based on natural movement trajectories.

The motion of the current prostheses is sequential and does not allow natural movements. In this work, complex natural motion patterns from a healthy upper limb were characterized in order to be emulated for a trans-humeral prosthesis with three degrees of freedom at the elbow. Firstly, it was necessary to define the prosthesis workspace, which means to establish a relationship using an artificial neural network (ANN), between the arm-forearm (3-D) angles allowed by the prosthesis, and its actuators length. The 3-D angles were measured between the forearm and each axis of the reference system attached at the elbow. Secondly, five activities of daily living (ADLs) were analyzed by means of the elbow flexion (EF), the forearm prono-supination (FPS) and the 3-D angles, from healthy subjects, by using a video-based motion analysis system. The 3-D angles were fed to the prosthesis model (ANN) in order to analyze which ADLs could be emulated by the prosthesis. As a result, a prosthesis kinematics approximation was obtained. In conclusion, in spite of the innovative mechanical configuration of the actuators, it was possible to carry out only three of the five ADLs considered. Future work will include improvement of the mechanical configuration of the prosthesis to have greater range of motion.