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Ultrasonic stimulation of the brain to enhance the release of dopamine – A potential novel treatment for Parkinson’s disease

Tian Xu, Xiaoxiao Lu, Danhong Peng, Gongdao Wang, Chen Chen, Wen Liu, Wei Wu, Timothy J. Mason

Ultrasonics Sonochemistry 2020 · 10.1016/j.ultsonch.2019.104955

rodentin vitro cellparkinsons diseasebehaviourhistology molecularcellular imaging

Abstract

Parkinson's disease (PD) is characterized by the decrease of dopamine (DA) production and release in the substantia nigra and striatum regions of the brain. Transcranial ultrasound has been exploited recently for neuromodulation of the brain in a number of fields. We have stimulated DA release in PC12 cells using low-intensity continuous ultrasound (0.1 W/cm 2 - 0.3 W/cm 2 , 1 MHz), 12 h after exposure at 0.2 W/cm 2 , 40 s, the amount of DA content eventually increased 78.5% (p = 0.004). After 10-day ultrasonic treatment (0.3 W/cm 2 , 5 min/d), the DA content in the striatum of PD mice model restored to 81.07% of the control (vs 43.42% in the untreated PD mice model). In addition to this the locomotion activity was restored to the normal level after treatment. We suggest that the low intensity ultrasound-induced DA release can be attributed to a combination of neuron regeneration and improved membrane permeability produced by the mechanical force of ultrasound. Our study indicates that the application of transcranial ultrasound applied below FDA limits, could provide a candidate for relatively safe and noninvasive PD therapy through an amplification of DA levels and the stimulation of dopaminergic neuron regeneration without contrast agents.

Abstract via europepmc.

Speciesmouse (C57BL/6J, MPTP model); rat PC12 cell line (in vitro)
Subjects12, 12, 12, 12, 12, 12, 12, 12, 12swept animals
Sessions per subject1, 5, 10swept
Randomisednot reported
Blindingnot reported
Sham / controlno treatment control
Auditory controlnot reported
Readout timingoffline
Anaesthesiaanaesthetised
Readoutsbehaviour, histology molecular, cellular imagingopen-field test; pole test; striatal dopamine content by HPLC-ECD; extracellular dopamine ELISA; tyrosine hydroxylase (TH) immunohistochemistry; H&E histology of skin/skull/brain; calcein/PI uptake by flow cytometry and confocal microscopy
Direction of effectexcitatoryLow-intensity ultrasound increased extracellular dopamine release from differentiated PC12 cells in vitro, and 10-day transcranial ultrasound treatment restored striatal dopamine content, TH-positive neuron numbers, and locomotor behaviour in MPTP-induced Parkinsonian mice toward control levels.
Adverse eventsobservedProlonged 0.3 W/cm2 exposure over 10 days produced slight superficial keratinization of the epidermis/hair follicles from probe friction; no pathological change was observed in HE staining of skull or whole brain in treated animals.

Exposures

Exposure 1: Continuous-wave ultrasound applied to differentiated PC12 cells in dish

Target: other — “differentiated PC12 cells (dish culture)
Device: other named manufacturer · Jiangsu Hanmei Science and Technology Co., Ltd. · HM-1

Pulse timing
Waveformcontinuous
Fundamental frequency (kHz)1,000✓✓
Pulse duration (ms)not applicable
Pulse repetition frequency (Hz)not applicable
Duty cycle (%)not applicable
Sonication duration (s)10, 20, 30, 40, 50, 60swept✓✓
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 applicable
In-situ pressure (kPa)not applicable
In-situ Isppa (W/cm²)not applicable
In-situ Ispta (W/cm²)not applicable
Ispta, domain unspecified (W/cm²)0.1, 0.2, 0.3swept✓✓
Protocol, in the paper’s words

Cells were irradiated over various periods of time (10, 20, 30, 40, 50 and 60 s) at intensities of 0.1, 0.2 and 0.3 W/cm2; control samples were run in parallel without sonication. The headline dopamine-release result used 0.2 W/cm2 for 40 s, assessed 12 h after exposure.

Exposure 2: Transcranial ultrasound to head of MPTP-induced PD mice (non-focused)

Target: whole brain or unfocused — “shaved scalp/head (low intensity non-focused ultrasound aimed at brain, striatum/SNc dopamine assessed)
Device: other named manufacturer · Jiangsu Hanmei Science and Technology Co., Ltd. · HM-1

Pulse timing
Waveformcontinuous
Fundamental frequency (kHz)1,000✓✓
Pulse duration (ms)not applicable
Pulse repetition frequency (Hz)not applicable
Duty cycle (%)not applicable
Sonication duration (s)300, 600, 900swept✓✓
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
Ispta, domain unspecified (W/cm²)0.1, 0.2, 0.3swept✓✓
Protocol, in the paper’s words

An L9(3^3) orthogonal design varied intensity I (0.1/0.2/0.3 W/cm2), daily exposure time t (5/10/15 min) and treatment period T (1/5/10 days) across 9 groups of 12 mice each. The headline restorative result used 0.3 W/cm2, 5 min/day, for 10 days, applied against the shaved scalp with ultrasound coupling gel.

Flags from extraction

  • n_subjectsPaper states group size (12 mice) and an L9(3^3) orthogonal design (9 groups) but never gives a single combined total exposed to ultrasound; group sizes listed as a 9-element list rather than summed.
  • exposures[0].targetPC12 is an immortalised rat pheochromocytoma cell line differentiated toward a neuron-like phenotype; no vocabulary target term matches a dish cell-line preparation, so 'other' was used.
  • exposures[1].targetThe transducer position on the head is not described relative to a specific brain region; the paper repeatedly calls the treatment 'low intensity non-focused ultrasound' aimed generally at the brain/striatum, so whole_brain_or_unfocused was used.
  • exposures[0].unspecified_domain.ispta_w_cm2The paper reports only 'intensity' without specifying ISPTA vs ISPPA or free-field vs in-situ; assigned to ispta_w_cm2/unspecified_domain because continuous-wave ISPPA=ISPTA and the FDA comparison discussed elsewhere in the paper is framed in terms of ISPTA limits.