Integrating Transcranial Ultrasound Stimulation with Exercise-Based Rehabilitation for Ischemic Stroke Recovery: A Preclinical Study

Ischemic stroke remains a leading cause of long-term disability worldwide, and effective neurorehabilitative strategies to enhance functional recovery are limited. This preclinical study compared noninvasive low-intensity pulsed ultrasound (LIPUS), aerobic exercise (AE) rehabilitation, and their combination in a mouse model of ischemic stroke. Transient focal cerebral ischemia was induced in male C57BL/6 mice using middle cerebral artery occlusion (MCAO). Male mice assigned to LIPUS-containing groups received LIPUS preconditioning for three consecutive days before MCAO, followed by two days of recovery and 14 days of post-stroke LIPUS, running wheel-based AE, or combined treatment. Functional recovery was assessed using open-field activity, neurological deficit scoring, and rotarod performance. Histological, immunohistochemical, TUNEL, and Western blot analyses were performed. MCAO caused marked motor and neurological impairments and brain tissue injury. LIPUS, AE, and their combination improved functional and histopathological outcomes, with the combined treatment showing greater effects for selected outcomes. All three interventions increased FNDC5 and BDNF and reduced TUNEL positivity. LIPUS and combined treatment, but not AE alone, increased SOD2 and catalase and reduced 8-OHdG immunoreactivity. These findings provide proof-of-concept in male mice to support further investigation of LIPUS combined with exercise rehabilitation for ischemic stroke recovery.