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Frequency dependent, reversible focused ultrasound suppression of evoked potentials in the reflex arc in an anesthetized animal

R. H. El Hassan, N. B. Lawand, E. D. Al‐Chaer, M. L. Khraiche

Journal of the Peripheral Nervous System 2022, 27, 271-282 · 10.1111/jns.12512

rodenthealthyemg mephistology molecular

Abstract

Objective To investigate the potential for noninvasive low frequency, low intensity ultrasound to suppress evoked potentials in the reflex arc neural pathway in anesthetized animal. Material and methods Single unit Electromyographic recordings of gastrocnemius muscle activity were obtained in response to electrical activation of the sciatic nerve, in anesthetized animal, with and without US stimulation. Reflex related potentials were evoked via electrical stimulation and low-intensity, low-frequency ultrasound was applied to the sciatic nerve. Electromyogram (EMG) was recorded from the gastrocnemius muscle and amplitude and area under the curve were analyzed to determine the effect of ultrasound stimulation on potentials in the reflex arc. Thermal imaging was used to assess thermal effects of US stimulation and immunohistochemical was performed post-US stimulation assess potential damage to the nerve. Results Our results show a drop in electromyogram amplitudes as high as 20%, and a drop in areas under the curve as high as 23%, with greater effects at lower frequencies (200 kHz) and lower acoustic intensities. The suppression of EMG scales with the magnitude of the electrical stimuli. Also, our results demonstrated transient reversibility of US suppression and our experiments eliminated thermal effects and mechanical and thermal damage. Conclusion The non-invasiveness of US stimulation and its inhibitory and reversible effects emphasize the potential of US as a therapeutic modality and clinical tool for suppression of neural potentials in the reflex arc such as the case for pain treatments. The study lays the ground for potential applications of US stimulation in pain treatment.

Abstract via europepmc.

Speciesrat (Sprague-Dawley)
Subjects5 animals
Sessions per subject1
Randomisedyes
Blindingnot reported
Sham / controlnone
Auditory controlnot reported
Readout timingboth
Anaesthesiaanaesthetised
Readoutsemg mep, histology molecularEMG amplitude and area-under-the-curve of gastrocnemius muscle evoked potentials in response to electrical sciatic nerve stimulation; immunohistochemistry (anti-Neurofilament 200, anti-S100) of the sciatic nerve
Direction of effectinhibitoryUS suppressed EMG amplitude and AUC of muscle-evoked potentials elicited by electrical sciatic-nerve stimulation; suppression was greater at the lower US frequency (200 kHz) and generally at lower acoustic intensities, persisted for 20 s post-stimulation, and recovered by ~30 min.
Adverse eventsnone observedTemperature change upon US stimulation was less than 0.3°C; immunofluorescence showed no degradation of neurofilament or damage to axons/Schwann cells following ultrasound stimulation.

Exposures

Exposure 1: 200 kHz continuous-wave US to sciatic nerve

Target: sciatic nerve — “sciatic nerve
Device: other named manufacturer · Mana Instruments · E1012-SU

Pulse timing
Waveformcontinuous
Fundamental frequency (kHz)200✓✓
Pulse duration (ms)not applicable
Pulse repetition frequency (Hz)not applicable
Duty cycle (%)not applicable
Sonication duration (s)10✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)not reported
Free-field Isppa (W/cm²)9.34, 4.16, 2.58swept✓✓
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
Protocol, in the paper’s words

Electrical stimulation was applied to the sciatic nerve for 20 s to evoke muscle activity, then continuous-wave US was applied simultaneously with electrical stimulation for 10 s, followed by 20 s of electrical stimulation alone to assess recovery. Each of 27 US/electrical parameter combinations plus 3 electrical-only controls was repeated three times in randomized order (90 recordings total per experiment), with 1 min rest between stimulations. During transducer characterization (not necessarily identical to the in-nerve exposure protocol), the transducer was described as operating with a 'burst duration 12 ms'.

Exposure 2: 500 kHz continuous-wave US to sciatic nerve

Target: sciatic nerve — “sciatic nerve
Device: other named manufacturer · Mana Instruments · E1012-SU

Pulse timing
Waveformcontinuous
Fundamental frequency (kHz)500✓✓
Pulse duration (ms)not applicable
Pulse repetition frequency (Hz)not applicable
Duty cycle (%)not applicable
Sonication duration (s)10✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)not reported
Free-field Isppa (W/cm²)11.1, 6.06, 5.04swept✓✓
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
Protocol, in the paper’s words

Electrical stimulation was applied to the sciatic nerve for 20 s to evoke muscle activity, then continuous-wave US was applied simultaneously with electrical stimulation for 10 s, followed by 20 s of electrical stimulation alone to assess recovery. Each of 27 US/electrical parameter combinations plus 3 electrical-only controls was repeated three times in randomized order (90 recordings total per experiment), with 1 min rest between stimulations. During transducer characterization (not necessarily identical to the in-nerve exposure protocol), the transducer was described as operating with a 'burst duration 12 ms'.

Exposure 3: 700 kHz continuous-wave US to sciatic nerve

Target: sciatic nerve — “sciatic nerve
Device: other named manufacturer · Mana Instruments · E1012-SU

Pulse timing
Waveformcontinuous
Fundamental frequency (kHz)700✓✓
Pulse duration (ms)not applicable
Pulse repetition frequency (Hz)not applicable
Duty cycle (%)not applicable
Sonication duration (s)10✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)not reported
Free-field Isppa (W/cm²)17.51, 5.65, 2.83swept✓✓
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
Protocol, in the paper’s words

Electrical stimulation was applied to the sciatic nerve for 20 s to evoke muscle activity, then continuous-wave US was applied simultaneously with electrical stimulation for 10 s, followed by 20 s of electrical stimulation alone to assess recovery. Each of 27 US/electrical parameter combinations plus 3 electrical-only controls was repeated three times in randomized order (90 recordings total per experiment), with 1 min rest between stimulations. During transducer characterization (not necessarily identical to the in-nerve exposure protocol), the transducer was described as operating with a 'burst duration 12 ms'.

Flags from extraction

  • exposures[0].timing.pulse_duration_msUltrasound characterization text states 'burst duration 12 ms' at 200/500/700 kHz, while the applied nerve-stimulation protocol states continuous-wave US applied for 10 s; unclear whether the 12 ms figure describes internal pulsing of the applied stimulus or is purely a calibration/characterization parameter, so pulse_duration_ms was left null rather than guessed.
  • exposures[0].timing.duty_cycle_pctPaper states waveform is a continuous sine wave but never gives an explicit duty-cycle percentage; left null rather than assuming 100%.
  • sham_typeNo inactive-transducer or sham-US condition was used; the only control condition was electrical stimulation alone (no US), classified here as 'none'.
  • in_situOnly water-tank (free-field) hydrophone-measured intensities are reported for the applied stimuli; no separate in-situ (derated/simulated/measured-in-tissue) values are given even though US was applied directly to an exposed nerve.