Integrating Transcranial Ultrasound Stimulation with Exercise-Based Rehabilitation for Ischemic Stroke Recovery: A Preclinical Study
Chang-Mu Chen, Ting-Yu Chang, Lin-Hwa Long, Cheng-Tien Wu, Shing-Hwa Liu
International Journal of Molecular Sciences 2026 · 10.3390/ijms27198498
Abstract
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.
Abstract via europepmc.
Exposures
Exposure 1: LIPUS 1 MHz to lateral motor cortex, pre- and post-MCAO
Target: motor cortex — “lateral motor cortex (M1/M2 region)”
Device: Mettler · Mettler Electronics · ME740 ✓
| Waveform | pulsed | |
|---|---|---|
| Fundamental frequency (kHz) | 1,000 | ✓✓✓ |
| Pulse duration (ms) | not reportedimplied by duty cycle ÷ PRF: 5 ms (not stated by the paper) | ⚑ |
| Pulse repetition frequency (Hz) | 100 | ✓✓✓ |
| Duty cycle (%) | 50 | ✓✓✓⚑ |
| Sonication duration (s) | 1,200 | ✓✓✓⚑ |
| Free-field pressure (kPa) | not reported | |
|---|---|---|
| Free-field Isppa (W/cm²) | not reported | |
| Free-field Ispta (W/cm²) | not reported | |
| In-situ estimate | not reported | |
| In-situ pressure (kPa) | not reported | |
| In-situ Isppa (W/cm²) | not reported | |
| In-situ Ispta (W/cm²) | not reported | |
| Ispta, domain unspecified (W/cm²) | 0.1 | ✓✓✓⚑ |
20 min per day for three consecutive days before MCAO, then daily for 14 days starting on postoperative day 3; non-focused collimated beam, ERA 1 cm2; output power 50%.
Flags from extraction
n_subjects— Seven groups of 9 mice; only the three LIPUS-containing groups were sonicated (listed as group sizes); total not stated.exposures[0].timing.duty_cycle_pct— Duty cycle 50% from Results text; Methods list 'output power: 50%'.exposures[0].timing.sonication_duration_s— 20 min exposure converted to 1200 s per daily session.exposures[0].timing.pulse_duration_ms— Pulse duration not stated.exposures[0].unspecified_domain.ispta_w_cm2— 100 mW/cm2 ISPTA converted to 0.1 W/cm2; domain not stated.anaesthesia— Anaesthesia stated for MCAO surgery only; not stated for LIPUS delivery.blinding— Behavioural and histology assessors blinded; those delivering interventions were not.exposures[0].target.label— Results text describes the target as the 'target cerebrovascular region'; Methods give lateral motor cortex coordinates.sham_type— Sham-operated mice and untreated MCAO group received no LIPUS.