Development of a Gene and Nucleic Acid Delivery System for Skeletal Muscle Administration via Limb Perfusion Using Nanobubbles and Ultrasound.

Strategies for gene and nucleic acid delivery to skeletal muscles have been extensively explored to treat Duchenne muscular dystrophy (DMD) and other neuromuscular diseases. Of these, effective intravascular delivery of naked plasmid DNA (pDNA) and nucleic acids into muscles is an attractive approach, given the high capillary density in close contact with myofibers. We developed lipid-based nanobubbles (NBs) using polyethylene-glycol-modified liposomes and an echo-contrast gas and found that these NBs could improve tissue permeability by ultrasound (US)-induced cavitation. Herein, we delivered naked pDNA or antisense phosphorodiamidate morpholino oligomers (PMOs) into the regional hindlimb muscle via limb perfusion using NBs and US exposure. pDNA encoding the luciferase gene was injected with NBs via limb perfusion into normal mice with application of US. High luciferase activity was achieved in a wide area of the limb muscle. DMD model mice were administered PMOs, designed to skip the mutated exon 23 of the dystrophin gene, with NBs via intravenous limb perfusion, followed by US exposure. The number of dystrophin-positive fibers increased in the muscles of mdx mice. Combining NBs and US exposure, which can be widely delivered to the hind limb muscles via the limb vein, could be an effective therapeutic approach for DMD and other neuromuscular disorders.

Sonoluminescence from ultra-high temperature and pressure cavitation produced by a narrow water jet.

This work developed a small-scale processing apparatus for ultra-high temperature and ultra-high-pressure cavitation (UTPC) incorporating a small diameter (0.1 mm) water jet nozzle. This instrumentation comprised a swirl flow nozzle (SFN) installed on the water jet nozzle so as to obtain UTPC from a multifunction cavitation (MFC) setup. Multi-bubble sonoluminescence (MBSL) assessments using two types of photon counting heads were employed to assess UTPC, MFC, ultrasonic cavitation (UC), water jet cavitation (WJC) and SFN-WJC. The SL intensity was found to increase in the order of SFN-WJC, WJC, UC, MFC to UTPC. Because UTPC produced the most intense emissions, this process evidently attained the highest processing temperature. Assuming a UC bubble temperature of 4000 K, the temperatures associated with UTPC, MFC and WJC were determined to be 5400-5900, 5300 and 3200-3300 K, respectively. The energy density of a single bubble during UTPC was calculated using the Rayleigh-Plesset and Planck equations for an initial bubble radius of 100 µm together with photon measurements from many bubbles and employing Planck's law. The highest SL intensity of UPTC is thought to exist due to the high energy density of UTPC. This research demonstrates that it is possible to increase the energy density of cavitation bubbles within a small reaction area.