SignificanceNeuromodulation devices are rapidly evolving for the treatment of neurological diseases and conditions. Injury from implantation or long-term use without obvious functional losses is often only detectable through terminal histology. New technologies are needed that assess the peripheral nervous system (PNS) under normal and diseased or injured conditions.AimWe aim to demonstrate an imaging and stimulation platform that can elucidate the biological mechanisms and impacts of neurostimulation in the PNS and apply it to the sciatic nerve to extract imaging metrics indicating electrical overstimulation.ApproachA sciatic nerve injury model in a 15-rat cohort was observed using a newly developed imaging and stimulation platform that can detect electrical overstimulation effects with polarization-sensitive optical coherence tomography. The sciatic nerve was electrically stimulated using a custom-developed nerve holder with embedded electrodes for 1 h, followed by a 1-h recovery period, delivered at above-threshold Shannon model k-values in experimental groups: sham control (SC, n = 5, 0.0 mA / 0 Hz), stimulation level 1 (SL1, n = 5, 3.4 mA / 50 Hz, and k = 2.57), and stimulation level 2 (SL2, n = 5, 6.8 mA / 100 Hz, and k = 3.17).ResultsThe stimulation and imaging system successfully captured study data across the cohort. When compared to a SC after a 1-week recovery, the fascicle closest to the stimulation lead showed an average change of +4 % / − 309 % (SL1/SL2) in phase retardation and −79 % / − 148 % in optical attenuation relative to SC. Analysis of immunohistochemistry (IHC) shows a +1 % / − 36 % difference in myelin pixel counts and −13 % / + 29 % difference in axon pixel counts, and an overall increase in cell nuclei pixel count of +20 % / + 35 % . These metrics were consistent with IHC and hematoxylin/eosin tissue section analysis.ConclusionsThe poststimulation changes observed in our study are manifestations of nerve injury and repair, specifically degeneration and angiogenesis. Optical imaging metrics quantify these processes and may help evaluate the safety and efficacy of neuromodulation devices.
SignificanceModern optical volumetric imaging modalities, such as optical coherence tomography (OCT), provide enormous information about the structure, function, and physiology of living tissue. Although optical imaging achieves lateral resolution on the order of the wavelength of light used, and OCT achieves axial resolution on a similar micron scale, tissue optical properties, particularly high scattering and absorption, limit light penetration to only a few millimeters. In addition, in vivo imaging modalities are susceptible to significant motion artifacts due to cardiac and respiratory function. These effects limit access to artifact-free optical measurements during peripheral neurosurgery to only a portion of the exposed nerve without further modification to the procedure.AimWe aim to improve in vivo OCT imaging during peripheral neurosurgery in small and large animals by increasing the amount of visualized nerve volume as well as suppressing motion of the imaged area.ApproachWe designed a nerve holder with embedded mirror prisms for peripheral nerve volumetric imaging as well as a specific beam steering strategy to acquire prism and direct view volumes in one session with minimal motion artifacts.ResultsThe axially imaged volumes from mirror prisms increased the OCT signal intensity by >22 dB over a 1.25-mm imaging depth in tissue-mimicking phantoms. We then demonstrated the new imaging capabilities in visualizing peripheral nerves from direct and side views in living rats and minipigs using a polarization-sensitive OCT system. Prism views have shown nerve fascicles and vasculature from the bottom half of the imaged nerve which was not visible in direct view.ConclusionsWe demonstrated improved OCT imaging during neurosurgery in small and large animals by combining the use of a prism nerve holder with a specifically designed beam scanning protocol. Our strategy can be applied to existing OCT imaging systems with minimal hardware modification, increasing the nerve tissue volume visualized. Enhanced imaging depth techniques may lead to a greater adoption of structural and functional optical biomarkers in preclinical and clinical medicine.
We have developed a novel nerve holder, imaging protocol and post-processing routine which enhance OCT imaging by accessing peripheral nerve from 3 sides. Two mirror prisms provide imaging access from sides of a peripheral nerve in addition to conventional volume acquisition from the top. Prism glass compensates for changed focusing and optical pathlength during nerve imaging from its sides Acquired prism sub-volumes and top-bottom imaged nerve sub-volume are merged in two-stage semi-automated postprocessing routine. We demonstrate successful application of this approach on tissue-mimicking phantoms, as well as in vivo rat sciatic nerve imaging.
Peripheral nerves connect and relay information between the central nervous system and its target organs. Small arteries traverse the epineurium and are responsible for supplying blood to the axons and cells within the nerves. Constriction or damage to these vessels can reduce perfusion leading to ischemic insults. Peripheral nerve electrostimulation has been approved for the treatment of epilepsy, depression and migraines, and is also being studied for the treatment of rheumatoid arthritis, Crohn’s disease, polycystic ovary syndrome, and type II diabetes. While the safety and efficacy of currently approved medical devices is well established, next generation devices may require novel stimulation parameters that pose additional risks. Therefore it is important to develop new methods to assess stimulation-induced nerve injury. To that end, we have begun to explore optical imaging based biomarkers, including optical coherence tomography angiography (OCT-A) to quantify changes in vascular morphology and blood flow during stimulation. We imaged the rat sciatic nerve in vivo with a 1300 nm OCT-A system. A 3-D printed nerve stabilizer with embedded platinum disc electrode was used to align the nerve for imaging during electrostimulation. Electrostimulation at either 40 or 400 µC/cm2 was applied for 1 hour. Images were acquired before, during and after stimulation. With higher electrostimulation parameters, blood vessels close to electrode site showed constriction. Immunohistochemical assessment was performed to correlate nerve injury to observed vascular changes. Optical imaging biomarkers have the potential to help assess the safety of novel electrodes and electrostimulation paradigms.
The peripheral nervous system (PNS) carries bidirectional information between the central nervous system and
distal organs. PNS stimulation has been widely used in medical devices for therapeutic indications, such as bladder
control and seizure cessation. Investigational uses of PNS stimulation include providing sensory feedback for
improved control of prosthetic limbs. While nerve safety has been well documented for stimulation parameters used
in marketed devices, novel PNS stimulation devices may require alternative stimulation paradigms to achieve
maximum therapeutic benefit. Improved testing paradigms to assess the safety of stimulation will expedite the
development process for novel PNS stimulation devices. The objective of this research is to assess peripheral nerve
vascular changes in real-time with optical coherence angiography (OCA). A 1300-nm OCA system was used to
image vasculature changes in the rat sciatic nerve in the region around a surface contacting single electrode. Nerves
and vasculature were imaged without stimulation for 180 minutes to quantify resting blood vessel diameter. Walking
track analysis was used to assess motor function before and 6 days following experiments. There was no significant
change in vessel diameter between baseline and other time points in all animals. Motor function tests indicated the
experiments did not impair functionality. We also evaluated the capabilities to image the nerve during electrical
stimulation in a pilot study. Combining OCA with established nerve assessment methods can be used to study the
effects of electrical stimulation safety on neural and vascular tissue in the periphery.
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