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Low-intensity (400 mW/cm 2 , 500 kHz) pulsed transcranial ultrasound preconditioning may mitigate focal cerebral ischemia in rats

Hangdao Li, Junfeng Sun, Daqu Zhang, Daryl Omire-Mayor, Peter A. Lewin, Shanbao Tong

Brain Stimulation 2017, 10, 695-702 · 10.1016/j.brs.2017.02.008

rodentstrokecerebral haemodynamicshistology molecular

Abstract

Background Preconditioning methods, which could increase tolerance of brain to subsequent ischemic injuries with a small dose of non-injury stimuli, have gained attention. Capitalizing on noninvasiveness and safety of ultrasound modality, the pulsed transcranial ultrasound stimulation (pTUS) approach may provide a novel treatment for patients with high risk of stroke. Objective This study's goal was to investigate whether the risk of stroke could be minimized or eliminated by prior exposure to low-intensity, pulsed transcranial ultrasound stimulation (pTUS). Methods Rats were randomly assigned to control (n = 12) and pTUS preconditioning (pTUS-PC) groups (n = 14). The animals in pTUS-PC group were exposed to transcranial ultrasound stimulation before the induction of photothrombotic stroke, whereas control animals were handled identically but without the ultrasound stimulation. Cerebral blood flow was monitored using laser speckle imaging in both groups during stroke induction, as well as 24 and 48 h after stroke, respectively. Also, infarct volumes and edema were measured at 48 h after stroke. Results pTUS-PC rats had smaller ischemic areas during stroke induction, and 24 and 48 h after the stroke, and smaller infarct volume (1.770 ± 0.169%) than the controls (3.215 ± 0.401%) (p Conclusion These results support the hypothesis that transcranial ultrasound stimulation applied before photothrombosis could provide neuroprotection by increasing the brain's tolerance to subsequently induced focal ischemic injury.

Abstract via europepmc.

Speciesrat (Sprague-Dawley)
Subjects14 animals
Sessions per subject1
Randomisedyes
Blindingnot reported
Sham / controlinactive transducer
Auditory controlnot reported
Readout timingboth
Anaesthesiaanaesthetised
Readoutscerebral haemodynamics, histology molecularLaser speckle imaging (LSI) of cerebral blood flow; TTC-stained brain sections for infarct volume and brain edema
Direction of effectinhibitoryThe pTUS parameters used were selected as 'suppressive' based on prior experimental/theoretical work intended to down-regulate brain metabolism/neural activity. pTUS preconditioning before photothrombotic stroke reduced ischemic areas, delayed hemodynamic compromise, and reduced infarct volume and brain edema at 48 h compared with controls.
Adverse eventsnot reportedSix rats died within 48 h after stroke induction (2/14 pTUS-PC, 4/12 control); the paper does not attribute these deaths to the ultrasound exposure, and mortality was numerically higher in the non-ultrasound control group.

Exposures

Exposure 1: pTUS preconditioning of primary somatosensory cortex before photothrombotic stroke

Target: primary somatosensory cortex — “the primary somatosensory cortex of the rat brain (right primary hind-limb somatosensory cortex)
Device: Olympus / Panametrics · Olympus NDT · V301-SU

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)500✓✓
Pulse duration (ms)0.5✓✓
Pulse repetition frequency (Hz)100✓✓
Duty cycle (%)not reportedpulse duration × PRF gives 5%
Sonication duration (s)10✓✓
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
Pressure, domain unspecified (kPa)589?
Ispta, domain unspecified (W/cm²)0.402?
Protocol, in the paper’s words

The ultrasound exposure was software-controlled: a 10 s single sequence (i.e., 1000 tone bursts) was repeated 180 times at a 10 s interval and delivered to the rat cortex before the induction of photothrombotic stroke. The pTUS-PC group was exposed to ultrasound treatment for 60 min total (chosen for comparison with a prior study). The control group was handled identically, including anaesthesia, but without the ultrasound stimulation.

Flags from extraction

  • n_subjects14 rats were enrolled in the pTUS-PC (ultrasound-exposed) group; 2 of these died before the 48 h endpoint and were excluded from analysis (enrolled n=14, analysed n=12). n_subjects reflects enrolled/exposed animals.
  • exposures[0].unspecified_domainThe paper states the peak pressure (589 kPa) and corresponding Ispta (402 mW/cm2) as 'delivered to the rat cortex' but does not explicitly state whether this was measured in water/free field, through the thinned skull, or simulated; classified as unspecified domain.
  • sham_typeThe paper states the control group was 'handled identically to the pTUS-PC group, with the exception of the ultrasound stimulation' but does not explicitly describe whether a transducer was placed without power (inactive_transducer) or simply omitted.