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Effects of external low intensity focused ultrasound on inflammatory markers in neuropathic pain

Abigail Hellman, Alicia Clum, Teresa Maietta, Adithya Srikanthan, Vraj Patel, Drishti Panse, Olivia Zimmerman, Paul Neubauer, Julia Nalwalk, Emery Williams, Goutam Ghoshal, Clif Burdette, Julie G. Pilitsis

Neuroscience Letters 2021, 757, 135977 · 10.1016/j.neulet.2021.135977

rodentchronic painbehaviourhistology molecular

Abstract

Background Changes in inflammatory cytokine levels contribute to the induction and maintenance of neuropathic pain. We have shown that external low intensity focused ultrasound (liFUS) reduces allodynia in a common peroneal nerve injury (CPNI). Here, we investigate an underlying mechanism of action for this treatment and measure the effect of liFUS on inflammatory markers. Methods Male rats were divided into four groups: CPNI/liFUS, CPNI/shamliFUS, shamCPNI/liFUS, and shamCPNI/shamliFUS. Mechanical nociceptive thresholds were measured using Von Frey filaments (VFF) to confirm the absence/presence of allodynia at baseline, after CPNI, and after liFUS. Commercial microarray and ELISA assays were used to assess cytokine expression in the treated L5 dorsal root ganglion (DRG) and dorsal horn (DH) tissue 24 and 72 h after liFUS. Results VFF thresholds were significantly reduced following CPNI in both groups that received the injury (p Conclusion We have demonstrated that liFUS modifies inflammatory cytokines in both DRG and DH in CPNI rats. These data provide evidence that liFUS, reverses the allodynic phenotype, in part, by altering inflammatory cytokine pathways.

Abstract via europepmc.

Speciesrat (Sprague-Dawley)
Subjects16, 16, 16, 16swept animals
Sessions per subject1
Randomisednot reported
Blindingnot reported
Sham / controlundescribed
Auditory controlnot reported
Readout timingoffline
Anaesthesiaanaesthetised
Readoutsbehaviour, histology molecularVon Frey filament mechanical nociceptive threshold testing; cytokine protein array (Proteome Profiler Rat Cytokine Array); IL6 and TNFa ELISA
Direction of effectinhibitoryliFUS to the L5 DRG significantly increased mechanical (Von Frey) withdrawal thresholds (reduced allodynia) only in CPNI animals, and decreased pro-inflammatory cytokines (TNFa, IL6, IL1b, CNTF, TIMP1) in DRG and dorsal horn while increasing anti-inflammatory IL10.
Adverse eventsnot reported

Exposures

Exposure 1: External liFUS to L5 dorsal root ganglion (DRG)

Target: dorsal root ganglion — “left L5 dorsal root ganglion (DRG)
Device: other named manufacturer · Acoustic MedSystems Inc

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)not reported
Pulse duration (ms)13✓✓
Pulse repetition frequency (Hz)38✓✓
Duty cycle (%)not reportedpulse duration × PRF gives 49.4%
Sonication duration (s)180✓✓
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
Protocol, in the paper’s words

A single external liFUS treatment (3 min at 8 W) was applied through the skin over the L5 DRG using an external landmark (posterior superior iliac spine), positioned so the beam travelled through soft tissue to the DRG without bony reflection; dose determined from a previously conducted dose-response curve. liFUS reverses allodynic responses in CPNI rats for approximately 3 days.

Flags from extraction

  • exposures[0].fundamental_frequency_khzPaper states 'a frequency of 38 Hz, pulse width of 13 ms, and period of 90 ns' - these values are internally inconsistent (a 90 ns period implies an ~11 MHz carrier, incompatible with a 13 ms pulse width), and 38 Hz is implausible as an ultrasound carrier frequency. 38 Hz and 13 ms were recorded as PRF and pulse duration respectively per the burst rule reasoning, but the fundamental (carrier) frequency is not clearly stated anywhere in the paper and is left not_reported.
  • exposures[0].timing.pulse_repetition_frequency_hzThe text labels 38 Hz as 'a frequency' rather than explicitly as PRF; interpreted as PRF because 38 Hz is not a plausible ultrasound carrier frequency, but this is an inference, not a direct statement.
  • sham_typeThe mechanism of sham liFUS delivery is not described in this paper (only named as 'shamliFUS' groups), so sham_type is set to 'other' per the rule for undescribed sham mechanisms.
  • n_subjectsVFF behavioral n=16/group is reported for each of the two liFUS-exposed groups (CPNI/liFUS, shamCPNI/liFUS) in each of the two separate cohorts (24h, 72h); no single total across cohorts is stated, so the four group sizes are listed.
  • exposures[0]No pressure, ISPPA, or ISPTA value is reported anywhere in the paper for the liFUS device; only total acoustic power (8 W) is given.