Precision mRNA Delivery via Ultrasound‐Controlled Release Perfluorocarbon Emulsions: An Innovative Ultrasound Theranostic Strategy with ¹⁹ F MRI Feasibility

Messenger RNA (mRNA) therapeutics provide promising opportunities in cardiovascular diseases. However, effective vascular mRNA delivery requires precise delivery, controlled release, and efficient transfection. To address these challenges, the study utilizes phase-change perfluorocarbon nanodroplets (PFC NDs) as a unique theranostic platform, integrating multimodal imaging with ultrasound-triggered mRNA release for enhanced vascular transfection. Lipid-coated PFC NDs are engineered using perfluoro-crown-ether (PFCE), perfluorohexane (PFH), and perfluoropentane (PFP) as core materials. These nanodroplets (200-300 nm) are optimized for mRNA loading and cellular uptake, exhibiting strong ultrasound contrast in tissue-mimicking phantoms and in vivo. Each PFC generated distinct fluorine-19 magnetic resonance imaging signals, allowing tri-spectral imaging capabilities. In vitro, PFH and PFP NDs increased enhanced green fluorescent protein mRNA transfection in CHO cells (p < 0.0001), with ultrasound stimulation further improving efficiency compared to unstimulated controls (p < 0.05). In vivo ultrasound-guided activation of PFH NDs resulted in higher mCherry protein expression in murine carotid arteries (p < 0.05), demonstrating site-specific gene therapy. This study establishes PFH NDs as an advanced theranostic platform for vascular mRNA delivery, integrating diagnostics and therapeutics into a single system. By leveraging ultrasound-responsive activation, these nanodroplets overcome existing delivery limitations, offering a new avenue for precision medicine in cardiovascular disease.