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Modulating the Inflammatory Reflex in Rats Using Low-Intensity Focused Ultrasound Stimulation of the Vagus Nerve

Kelsey M. Wasilczuk, Kelsey C. Bayer, Jesse P. Somann, Gabriel O. Albors, Jennifer Sturgis, L. Tiffany Lyle, J. Paul Robinson, Pedro P. Irazoqui

Ultrasound in Medicine & Biology 2019, 45, 481-489 · 10.1016/j.ultrasmedbio.2018.09.005

rodentsystemic inflammationhistology molecularautonomic physiology

Abstract

Tumor necrosis factor α (TNF-α) is linked to several chronic inflammatory diseases. Electrical vagus nerve stimulation reduces serum TNF-α levels but may cause chronic nerve damage and requires surgery. Alternatively, we proposed focused ultrasound stimulation of the vagus nerve (uVNS), which can be applied non-invasively. In this study, we induced an inflammatory response in rats using lipopolysaccharides (LPS) and collected blood to analyze the effects of uVNS on cytokine concentrations. We applied one or three 5-min pulsed focused ultrasound stimulation treatments to the vagus nerve (250 kHz, I SPPA = 3 W/cm 2 ). Animals receiving a single ultrasound application had an average reduction in TNF-α levels of 19%, similar to the 16% reduction observed in electrically stimulated animals. With multiple applications, uVNS therapy statistically reduced serum TNF-α levels by 73% compared with control animals without any observed damage to the nerve. These findings suggest that uVNS is a suitable way to attenuate TNF-α levels.

Abstract via europepmc.

Speciesrat (Sprague-Dawley)
Subjects11, 8, 4swept animals
Sessions per subject1, 3swept
Randomisednot reported
Blindingsingle
Sham / controlnone
Auditory controlnot reported
Readout timingoffline
Anaesthesiaanaesthetised
Readoutshistology molecular, autonomic physiologyserum cytokine panel (LEGENDplex 13-plex; TNF-alpha primary outcome); vagus nerve histology (H&E)
Direction of effectinhibitoryuVNS reduced serum TNF-alpha levels (single application ~19% reduction, triple application ~73% reduction vs. no-stimulation controls), interpreted as activation of the cholinergic anti-inflammatory reflex; direct neural activation (action potentials) was not recorded, and the effect was abolished by distal efferent vagotomy.
Adverse eventsnone observedHistology of 8 vagus nerve specimens showed no significant lesions except mild, clinically insignificant multifocal neutrophil infiltration of the perineurium in one animal; temperature increased on average to a maximum of 2C over the 5-min stimulation.

Exposures

Exposure 1: Focused ultrasound vagus nerve stimulation (uVNS)

Target: vagus nerve — “left cervical vagus nerve (surgically exposed)
Device: Sonic Concepts · Sonic Concepts · H-115

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)250✓✓
Pulse duration (ms)4✓✓
Pulse repetition frequency (Hz)10✓✓
Duty cycle (%)not reportedpulse duration × PRF gives 4%
Sonication duration (s)300✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)not reported
Free-field Isppa (W/cm²)3✓✓
Free-field Ispta (W/cm²)1.2✓✓
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

Focused ultrasound was delivered for 5 min per stimulation session, applied to the surgically exposed left vagus nerve via a coupling cone and ultrasound gel. Animals received either a single stimulation (u1VNS) or three stimulations (u3VNS, u3Vx) at t=30, 90 and 150 min relative to LPS injection.

Flags from extraction

  • n_subjectsPaper never states a combined total for ultrasound-exposed animals; group sizes for the three uVNS groups (u1VNS=11, u3VNS=8, u3Vx=4) are listed instead of summed.
  • exposures[0].free_field.isppa_w_cm2Intensity was measured with a hydrophone in water at the focus but described in the text as the intensity 'applied to the vagus nerve'; since the nerve was surgically exposed and coupled with gel (no skull path), this was treated as a free-field measurement, but the paper does not explicitly state the domain.
  • randomisedPaper does not state whether animals were randomly allocated to groups.