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Protective Effect of Low-Intensity Pulsed Ultrasound on Memory Impairment and Brain Damage in a Rat Model of Vascular Dementia

Sin-Luo Huang, Chi-Wei Chang, Yi-Hsuan Lee, Feng-Yi Yang

Radiology 2017, 282, 113-122 · 10.1148/radiol.2016160095

rodenthealthyvascular dementiapetbehaviourhistology molecular

Abstract

Purpose To investigate the neuroprotective effects of low-intensity pulsed (LIP) ultrasound on memory impairment and central nervous system injury in a rat model of vascular dementia. Materials and Methods All animal experiments were approved by the animal care and use committee and adhered to experimental animal care guidelines. A 1.0-MHz focused ultrasound transducer was used to stimulate the brain noninvasively with 50-msec bursts at a 5% duty cycle, repetition frequency of 1 Hz, and spatial peak temporal average intensity of 528 mW/cm 2 . LIP ultrasound treatment was performed daily with triple sonications in each hemisphere. The duration of each sonicaton was 5 minutes, with a 5-minute interval between each sonication. Permanent bilateral common carotid artery occlusion (BCCAO) was used as a model of vascular dementia. After 2 weeks of LIP ultrasound, neuroprotective effects of LIP ultrasound were evaluated with behavioral analysis, including the passive avoidance task and elevated plus maze. Myelin content was detected with carbon 11 ( 11 C) Pittsburgh compound B (PIB). Brain sections were stained with hematoxylin-eosin and Luxol fast blue. Two-way analysis of variance and Student t test were used for statistical analyses, with a significance level of .05. Results Protein expressions of brain-derived neurotrophic factor (BDNF) in the BCCAO rats treated with LIP ultrasound were significantly higher than those in BCCAO rats (1.1 ± 0.0 vs 0.8 ± 0.1, P 11 C PIB accumulation in the BCCAO rats treated with LIP ultrasound was significantly (P © RSNA, 2016.

Abstract via europepmc.

Speciesrat (Sprague-Dawley)
Subjectsnot reported animals
Sessions per subject14
Randomisedyes
Blindingsingle
Sham / controlnone
Auditory controlnot reported
Readout timingoffline
Anaesthesianot reported
Readoutspet, behaviour, histology molecularWestern blot for BDNF; immunohistochemistry for BDNF and myelin; 11C-Pittsburgh compound B (PIB) micro-PET imaging; passive avoidance task; elevated plus maze; hematoxylin-eosin and Luxol fast blue staining
Direction of effectnot assessedLIP ultrasound stimulation increased BDNF protein levels and myelin content (Luxol fast blue staining) and improved learning/memory (passive avoidance, elevated plus maze) in BCCAO rats compared with untreated BCCAO rats; no direct electrophysiological measure of excitation/inhibition was made.
Adverse eventsnone observedNo lesions were detected in sham rats and healthy brains after LIP ultrasound treatment (Fig 4a).

Exposures

Exposure 1: LIP ultrasound treatment of the brain (sham/healthy and BCCAO rats)

Target: brain — “the desired region (3.0 mm posterior and 2.5 mm lateral to the bregma) of the brain
Device: Olympus / Panametrics · Panametrics · A392S

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)1,000✓✓
Pulse duration (ms)50✓✓
Pulse repetition frequency (Hz)1✓✓
Duty cycle (%)5pulse duration × PRF gives 5%✓✓
Sonication duration (s)300✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)not reported
Free-field Isppa (W/cm²)not reported
Free-field Ispta (W/cm²)0.528✓✓
In-situ estimatenot reported
In-situ pressure (kPa)not reported
In-situ Isppa (W/cm²)not reported
In-situ Ispta (W/cm²)not reported
Protocol, in the paper’s words

Each rat hemisphere was treated with LIP ultrasound daily with triple sonications. The duration of each sonication was 5 minutes, with a 5-minute interval between each sonication. Animals in the LIP ultrasound group were treated with LIP ultrasound daily for 14 days (in the BCCAO+LIP ultrasound group, starting 2 weeks after BCCAO surgery).

Flags from extraction

  • n_subjectsSample sizes vary by experiment/readout (n=3 for histology, n=4 for biochemical/PET biodistribution, n=8 for behavioural tests); no single total of ultrasound-exposed animals is stated.
  • blindingPaper states quantification of BDNF/myelin staining was performed 'in a blinded manner' (outcome assessment blinding); no statement about blinding of treatment allocation or behavioural testing.
  • anaesthesiaIsoflurane anaesthesia is described for BCCAO surgery and PET imaging, but the paper does not state whether animals were anaesthetised during the daily LIP ultrasound stimulation sessions themselves.