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Transcranial Focused Ultrasound Stimulation Improves Neurorehabilitation after Middle Cerebral Artery Occlusion in Mice

Jixian Wang, Guofeng Li, Lidong Deng, Muyassar Mamtilahun, Lu Jiang, Weibao Qiu, Hairong Zheng, Junfeng Sun, Qing Xie, Guo-Yuan Yang

Aging and disease 2021, 12, 50 · 10.14336/ad.2020.0623

rodentstrokebehaviourhistology molecularother

Abstract

Transcranial focused ultrasound stimulation (tFUS) regulates neural activity in different brain regions in humans and animals. However, the role of ultrasound stimulation in modulating neural activity and promoting neurorehabilitation in the ischemic brain is largely unknown. In the present study, we explored the effect of tFUS on neurological rehabilitation and the underlying mechanism. Adult male ICR mice (n=42) underwent transient middle cerebral artery occlusion. One week after brain ischemia, low frequency (0.5 MHz) tFUS was applied to stimulate the ischemic hemisphere of mice for 7 consecutive days (10 minutes daily). Brain infarct volume, neurobehavioral tests, microglia activation, IL-10 and IL-10R levels were further assessed for up to 14 days. We found that the brain infarct volume was significantly reduced in the tFUS treated mice compared to that in the non-treated mice ( p p p <0.05). We concluded that tFUS served as a unique technique to promote neurorehabilitation after brain ischemia by promoting microglia polarization and further regulating IL-10 signaling in the ischemic brain.

Abstract via europepmc.

Speciesmouse (ICR)
Subjects18 animals
Sessions per subject7
Randomisednot reported
Blindingsingle
Sham / controlnone
Auditory controlnot reported
Readout timingoffline
Anaesthesiaanaesthetised
Readoutsbehaviour, histology molecular, otherNeurological severity score (NSS), elevated body swing test (EBST), corner test; cresyl violet infarct/atrophy volume; IBA1/CD16-32/arginase/IL-10R immunostaining; flow cytometry (FACS) for M1/M2 microglia; Western blot for IL-10R; RT-PCR for IL-10 and IL-10R mRNA
Direction of effectnot assessedStudy assessed therapeutic/behavioural and anti-inflammatory outcomes (reduced infarct volume, improved NSS/EBST/corner test, increased M2 microglia and IL-10/IL-10R) rather than excitatory/inhibitory direction of neural activity.
Adverse eventsnot reported

Exposures

Exposure 1: tFUS to ischemic hemisphere after MCAO

Target: cerebral cortex — “ischemic hemisphere (selected region of the cortex)
Device: custom-built · Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)500✓✓
Pulse duration (ms)0.5✓✓
Pulse repetition frequency (Hz)1,000✓✓
Duty cycle (%)not reportedpulse duration × PRF gives 50%
Sonication duration (s)0.3✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)not reported
Free-field Isppa (W/cm²)not reported
Free-field Ispta (W/cm²)not reported
In-situ estimatenot reported
In-situ pressure (kPa)not reported
In-situ Isppa (W/cm²)not reported
In-situ Ispta (W/cm²)not reported
Isppa, domain unspecified (W/cm²)0.12✓✓
Protocol, in the paper’s words

tFUS was delivered with a portable ultrasound stimulation system to the selected cortical region of the ischemic hemisphere; the stimulation sequence used a toneburst-duration (TBD), pulse repetition frequency (PRF), sonication duration (SD) and 2.7 s inter-stimuli interval (ISI) as illustrated in Fig. 2D. Beginning 7 days after MCAO, ultrasound stimulation was performed for 10 minutes per day over 7 consecutive days, totaling 360 trials of ultrasound stimulation.

Flags from extraction

  • exposures[0].unspecified_domain.isppa_w_cm2Paper does not state whether the reported Isppa (120 mW/cm2) is a free-field or in-situ value; placed in unspecified_domain.
  • exposures[0].timing.duty_cycle_pctDuty cycle is not stated explicitly; arithmetic from pulse duration (0.5 ms) and PRF (1000 Hz) would suggest 50% but this is not computed per instructions.
  • sham_typeOnly an untreated MCAO-alone control group is described; no sham ultrasound (e.g. inactive transducer) arm was used.