The development of mechanically formed stable nanobubbles intended for sonoporation-mediated gene transfection

Abstract

In this study, stable nano-sized bubbles (nanobubbles [NBs]) were produced using the mechanical agitation method in the presence of perfluorocarbon gases. NBs made with perfluoropropane had a smaller size (around 400 nm) compared to that of those made with perfluorobutane or nitrogen gas. The lipid concentration in NBs affected both their initial size and post-formulation stability. NBs formed with a final lipid concentration of 0.5 mg/ml tended to be more stable, having a uniform size distribution for 24 h at room temperature and 50 h at 4 C. In vitro gene expression revealed that NBs/pDNA in combination with ultrasound (US) irradiation had significantly higher transfection efficacy in colon C26 cells. Moreover, for in vivo gene transfection in mice left limb muscles, there was notable local transfection activity by NBs/pDNA when combined with US irradiation. In addition, the aged NBs kept at room temperature or 4 C were still functional at enhancing gene transfection in mice. We succeeded in preparing stable NBs for efficient in vivo gene transfection, using the mechanical agitation method.

Cavitation-threshold Determination and Rheological-parameters Estimation of Albumin-stabilized Nanobubbles

“…There are numerous applications for which NBs provide interesting results, notably drug delivery to various targets: tumor 50 – 58 , nerve 59 , retina 60 , vascular tissues 61 , brain blood barrier 62 . They can also be used to perform gene transfection 63 – 68 . Finally, they showed interesting features for imaging applications 69 – 73 and theragnostic modalities 74 – 77 .…”

Advanced physical techniques for gene delivery based on membrane perforation

“…Too high or too low concentration of microbubbles can reduce the transfection efficiency of sonoporation. Recently, nanobubbles as an effective contrast agent are also used for sonoporation-mediated gene transfection by some scholar (Nishimura et al., 2017 ; Abdalkader et al., 2017 ). In addition, other nanocarriers, such as polymeric nanoparticles, polymeric micelles, liposomes or nanoemulsions, are also combined with sonoporation to enhance the transfection efficiency (Husseini & Pitt, 2008 ).…”

Evaluation of the targeted delivery of 5-fluorouracil and ascorbic acid into the brain with ultrasound-responsive nanobubbles

“…L-Ascorbic acid was purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan) and dissolved in water at 10 mg/mL. BLs composed of 1,2-Distearoyl-sn-glycero-3-phosphocholine (DSPC) (Avanti Polar Lipids, Inc., Alabaster, Alabama, USA) and 1,2-distearoyl-sn-glycero-3-phosphatidylethanolaminemethoxy-polyethyleneglycol (DSPE-PEG (2k)-OMe) (NOF Corporation, Tokyo, Japan) (94:6 (m/m)) were prepared according to previous reports at 1 mg/mL in terms of their lipids [16,17,20,21]. To encapsulate the perfluoropropane gas into BLs, vials were vortexed in a sonicated bath (BRANSONIC B1200, Emerson Electric Co., St. Louis, MO, USA).…”