Scalable production of siRNA-encapsulated extracellular vesicles for the inhibition of KRAS-mutant cancer using acoustic shock waves.

Extracellular vesicles (EVs) have emerged as a potential delivery vehicle for nucleic-acid-based therapeutics, but challenges related to their large-scale production and cargo-loading efficiency have limited their therapeutic potential. To address these issues, we developed a novel "shock wave extracellular vesicles engineering technology" (SWEET) as a non-genetic, scalable manufacturing strategy that uses shock waves (SWs) to encapsulate siRNAs in EVs. Here, we describe the use of the SWEET platform to load large quantities of KRASG12C-targeting siRNA into small bovine-milk-derived EVs (sBMEVs), with high efficiency. The siRNA-loaded sBMEVs effectively silenced oncogenic KRASG12C expression in cancer cells; they inhibited tumour growth when administered intravenously in a non-small cell lung cancer xenograft mouse model. Our study demonstrates the potential for the SWEET platform to serve as a novel method that allows large-scale production of cargo-loaded EVs for use in a wide range of therapeutic applications.

High-resolution photoacoustic/ultrasound imaging of the porcine stomach wall: an ex vivo feasibility study.

Photoacoustic (PA) imaging has become invaluable in preclinical and clinical research. Endoscopic PA imaging in particular has been explored as a noninvasive imaging modality to view vasculature and diagnose cancers in the digestive system. However, these feasibility studies are still limited to rodents or rabbits. Here, we develop a fully synchronized simultaneous ultrasound and photoacoustic microscopy system using two spectral bands (i.e., the visible and near-infrared) in both optical- and acoustic-resolution modes. We investigate the feasibility of imaging gastric vasculature in an ex vivo porcine model. The entire gastric wall, including the mucosa, submucosa, muscularis propria, and serosa, was excised from fresh porcine stomachs immediately followed by ultrasound and PA imaging being performed within a few hours of sacrifice. PA images of the mucosal vasculature were obtained at depths of 1.90 mm, which is a clinically significant accomplishment considering that the average thickness of the human mucosa is 1.26 mm. The layer structure of the stomach wall could be clearly distinguished in the overlaid PA and US images. Because gastric cancer starts from the mucosal surface and infiltrates into the submucosa, PA imaging can cover a clinically relevant depth in early gastric cancer diagnosis. We were able to detect mucosal vasculature in the entire mucosal layer, suggesting the potential utility of combined PA/US imaging in gastroenterology.

The role of the acoustic radiation force in color Doppler twinkling artifacts.

PURPOSE: The aim of this experimental study was to evaluate whether the acoustic radiation force (ARF) is a potential source of twinkling artifacts in color Doppler images. METHODS: Color Doppler images were obtained using a clinical ultrasonic scanner (Voluson e, GE Healthcare) for a high contrast (+15 dB) circular scattering phantom at pulse repetition frequencies (PRFs) ranging from 0.1 to 13 kHz. Ultrasound transmissions resulting in ARF were measured using a hydrophone at the various PRFs considered. The influence of ARF on the appearance of twinkling colors was examined via the common parameter PRF. This methodology is based on the fact that alternating positive and negative Doppler shifts induced by the ARF are centered at a PRF twice the maximum Doppler frequency on the color scale bar, whereas the twinkling color aliasing is expected to remain similar regardless of PRF. RESULTS: Color twinkling artifacts were observed to be most conspicuous at the lowest PRF of 0.1 kHz. The extent of twinkling rapidly decreased as the PRF increased, eventually disappearing when the PRF ≥0.6 kHz. The measured ultrasound transmissions, however, were found to be insensitive to the PRF, and therefore it can be inferred that the PRF was insensitive to the ARF. CONCLUSION: Based on our experimental observations, the ARF may not be a source of color Doppler twinkling artifacts.