Holographic focused ultrasound for spatially precise multi-target therapy in ischemic stroke
Noninvasive delivery of mesenchymal stromal cells (MSCs) and extracellular vesicles (EVs) to the peri-infarct zone is highly desired but challenging in ischemic stroke, due to the difficulties in achieving precise spatial targeting across diverse infarct topographies and circumventing the blood-brain barrier (BBB). Here, we introduce a rodent hologram-assisted multi-target BBB opening (HaMBO) platform using a 3D-printed hologram coupled to a single-element ultrasound transducer. The designed holograms accurately tailored multiple foci to the arc-shaped infarct border, as validated by numerical simulations and hydrophone measurements. HaMBO enabled precise tracer delivery and did not cause detectable behavioral, histological, or transcriptomic abnormalities. A single two-minute HaMBO achieved efficient peri-infarct delivery of MSCs and EVs injected intravenously, significantly reducing brain atrophy (~70%), promoting angiogenesis (~2 folds) and neurogenesis (~4 folds), and improving long-term behavioral recovery in a transient focal cerebral ischemia mouse model. Moreover, 3D simulations in a human skull model confirmed the ability of acoustic holography to generate arbitrarily shaped ultrasound fields for targeting diverse ischemic stroke patterns. These findings establish HaMBO as a safe, efficient, low-cost, and customizable platform for personalized therapeutic delivery in ischemic stroke, with broad translational potential for other neurological disorders requiring multi-target intervention.