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Noninvasive Ultrasound Retinal Stimulation for Vision Restoration at High Spatiotemporal Resolution

Xuejun Qian, Gengxi Lu, Biju B. Thomas, Runze Li, Xiaoyang Chen, K. Kirk Shung, Mark Humayun, Qifa Zhou

BME Frontiers 2022, 2022 · 10.34133/2022/9829316

rodenthealthyretinal degenerationinvasive electrophysiologyhistology molecular

Abstract

Objective . Retinal degeneration involving progressive deterioration and loss of function of photoreceptors is a major cause of permanent vision loss worldwide. Strategies to treat these incurable conditions incorporate retinal prostheses via electrically stimulating surviving retinal neurons with implanted devices in the eye, optogenetic therapy, and sonogenetic therapy. Existing challenges of these strategies include invasive manner, complex implantation surgeries, and risky gene therapy. Methods and Results . Here, we show that direct ultrasound stimulation on the retina can evoke neuron activities from the visual centers including the superior colliculus and the primary visual cortex (V1), in either normal-sighted or retinal degenerated blind rats in vivo . The neuron activities induced by the customized spherically focused 3.1 MHz ultrasound transducer have shown both good spatial resolution of 250 μ m and temporal resolution of 5 Hz in the rat visual centers. An additional customized 4.4 MHz helical transducer was further implemented to generate a static stimulation pattern of letter forms. Conclusion . Our findings demonstrate that ultrasound stimulation of the retina in vivo is a safe and effective approach with high spatiotemporal resolution, indicating a promising future of ultrasound stimulation as a novel and noninvasive visual prosthesis for translational applications in blind patients.

Abstract via europepmc.

Speciesrat (Long-Evans and RCS strains)
Subjects26 animals
Sessions per subjectnot reported
Randomisednot reported
Blindingnot reported
Sham / controlinactive transducer
Auditory controlother
Readout timingonline
Anaesthesiaanaesthetised
Readoutsinvasive electrophysiology, histology molecularmultiunit neural recordings (32- and 56-channel MEA) from superior colliculus and visual cortex; retinal histology (H&E) for safety assessment
Direction of effectexcitatoryUltrasound stimulation of the retina evoked spike activity in the superior colliculus and visual cortex comparable to light-evoked responses in normal-sighted rats, and restored evoked responses in blind RCS rats where light stimulation failed; response amplitude and duration increased with US intensity and duration.
Adverse eventsnone observedHistology of stimulated retinas from normal-sighted and retinal degenerative rats showed the retina structure remained intact with no obvious changes in retinal architecture even after 2 hours of US stimulation.

Exposures

Exposure 1: Retinal US stimulation, 3.1 MHz spherically focused transducer (parametric study)

Target: retina — “retina
Device: custom-built · custom (USC Dept. of Biomedical Engineering) · custom 3.1 MHz spherically focused single-element transducer (DL-47 piezo, Del-Piezo Specialties)

Pulse timing
Waveformcontinuous, pulsed
Fundamental frequency (kHz)3,100✓✓
Pulse duration (ms)not reported
Pulse repetition frequency (Hz)not reported
Duty cycle (%)100, 30, 50, 70swept✓✓
Sonication duration (s)0.001, 0.2swept✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)1,290, 3,370swept✓✓
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

The default US sequence was a continuous wave at 100% duty cycle with a 6-second frame interval between stimulations to allow full neuronal recovery. Driving voltage (hence free-field NPP, 1.29-3.37 MPa) and US duration (stated as 1-200 ms in Results, 50-200 ms in Methods) were each swept to assess their effect on evoked SC/VC responses; pulsed-mode sequences with 30%, 50%, or 70% duty cycle were also compared against the continuous mode at matched time-averaged energy. Temporal resolution was assessed with 10-ms, 2.83 MPa NPP stimuli delivered at frame rates of 1, 2, 5, and 10 Hz over 20-second trains. Prior to each stimulation series, the beam was directed away from the eye as an off-target control.

Exposure 2: Static letter-pattern generation, 4.4 MHz helical transducer

Target: retina — “retina (letter-pattern generation)
Device: custom-built · custom (USC Dept. of Biomedical Engineering) · custom 4.4 MHz helical transducer (1-3 composite DL-48, Del-Piezo Specialties)

Pulse timing
Waveformnot reported
Fundamental frequency (kHz)4,400✓✓
Pulse duration (ms)not reported
Pulse repetition frequency (Hz)not reported
Duty cycle (%)100✓✓
Sonication duration (s)not reported
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-element helical transducer (4.4 MHz, 4 mm outer/2 mm inner diameter, 10 mm focal length, 1.2 mm vertical separation) was used to generate a static acoustic beam pattern in the shape of the letter 'C' on the retina, matched to the hydrophone-measured water-condition field pattern, and MEA recordings in SC confirmed a corresponding response pattern.

Flags from extraction

  • exposures[0].timing.sonication_duration_sResults text states US duration was explored 'from 1 ms to 200 ms', while the Methods section states 'a test ranging from 50 ms to 200 ms with an interval of 50 ms'; the Results range was recorded here and the discrepancy is flagged.
  • exposures[0].timing.waveformDefault sequence was continuous wave (100% duty cycle), but the study also compared pulsed sequences at 30/50/70% duty cycle without stating pulse duration or PRF for these; recorded as a list of waveform types.
  • exposures[0].timing.pulse_repetition_frequency_hzPRF was not stated numerically for the pulsed-mode duty-cycle comparison conditions.
  • n_subjects26 rats were investigated in total across all sub-experiments; not all were necessarily exposed to identical parameters (see Supplementary Table 1 for the breakdown, not available in the extracted text).