Donghak Byun

Fabrication of a flexible penetrating microelectrode array for use on curved surfaces of neural tissues

Conventionally, invasive neural microelectrodes for recording neuronal signals or stimulating the nervous system have been fabricated based on silicon substrate mainly due to well-established manufacturing processes. However, these silicon-based microelectrode devices have an issue of mechanical stability caused by the absence of flexibility when implanted onto curved surfaces of tissues. In this paper, a flexible and penetrating microelectrode array, a hybrid structure composed of silicon and elastomer, was devised and fabricated by bulk micromachining technologies. The structure uses individual silicon needles as independent electrodes in a square array and polydimethysiloxane (PDMS) as a base to support the needles. The dimensions of the electrode array and the needles are adjustable, depending on the number of needles, the pitch between the needles and the targeted penetration depth of the neural tissue. For mechanical characterization, the adhesion between PDMS and silicon was evaluated and the flexibility and integrity of the fabricated structure were investigated through flexural test and insertion test. Also, the electrochemical impedance spectroscopy of the electrodes was measured. The results suggest that the proposed microelectrode array is promising for use in neuronal recording and stimulation over curved surfaces such as cortical surface and peripheral nerves with larger curvatures.

Zebrafish needle EMG: a new tool for high-throughput drug screens

Zebrafish models have recently been highlighted as a valuable tool in studying the molecular basis of neuromuscular diseases and developing new pharmacological treatments. Needle electromyography (EMG) is needed not only for validating transgenic zebrafish models with muscular dystrophies (MD), but also for assessing the efficacy of therapeutics. However, performing needle EMG on larval zebrafish has not been feasible due to the lack of proper EMG sensors and systems for such small animals. We introduce a new type of EMG needle electrode to measure intramuscular activities of larval zebrafish, together with a method to hold the animal in position during EMG, without anesthetization. The silicon-based needle electrode was found to be sufficiently strong and sharp to penetrate the skin and muscles of zebrafish larvae, and its shape and performance did not change after multiple insertions. With the use of the proposed needle electrode and measurement system, EMG was successfully performed on zebrafish at 30 days postfertilization (dpf) and at 5 dpf. Burst patterns and spike morphology of the recorded EMG signals were analyzed. The measured single spikes were triphasic with an initial positive deflection, which is typical for motor unit action potentials, with durations of ∼10 ms, whereas the muscle activity was silent during the anesthetized condition. These findings confirmed the capability of this system of detecting EMG signals from very small animals such as 5 dpf zebrafish. The developed EMG sensor and system are expected to become a helpful tool in validating zebrafish MD models and further developing therapeutics.

Effect of Bone Cement Volume and Stiffness on Occurrences of Adjacent Vertebral Fractures after Vertebroplasty

Objective The purpose of this study is to find the optimal stiffness and volume of bone cement and their biomechanical effects on the adjacent vertebrae to determine a better strategy for conducting vertebroplasty. Methods A three-dimensional finite-element model of a functional spinal unit was developed using computed tomography scans of a normal motion segment, comprising the T11, T12 and L1 vertebrae. Volumes of bone cement, with appropriate mechanical properties, were inserted into the trabecular core of the T12 vertebra. Parametric studies were done by varying the volume and stiffness of the bone cement. Results When the bone cement filling volume reached 30% of the volume of a vertebral body, the level of stiffness was restored to that of normal bone, and when higher bone cement exceeded 30% of the volume, the result was stiffness in excess of that of normal bone. When the bone cement volume was varied, local stress in the bony structures (cortical shell, trabecular bone and endplate) of each vertebra monotonically increased. Low-modulus bone cement has the effect of reducing strain in the augmented body, but only in cases of relatively high volumes of bone cement (>50%). Furthermore, varying the stiffness of bone cement has a negligible effect on the stress distribution of vertebral bodies. Conclusion The volume of cement was considered to be the most important determinant in endplate fracture. Changing the stiffness of bone cement has a negligible effect on the stress distribution of vertebral bodies.

Zebrafish as an animal model in epilepsy studies with multichannel EEG recordings

Despite recent interest in using zebrafish in human disease studies, sparked by their economics, fecundity, easy handling, and homologies to humans, the electrophysiological tools or methods for zebrafish are still inaccessible. Although zebrafish exhibit more significant larval–adult duality than any other animal, most electrophysiological studies using zebrafish are biased by using larvae these days. The results of larval studies not only differ from those conducted with adults but also are unable to delicately manage electroencephalographic montages due to their small size. Hence, we enabled non-invasive long-term multichannel electroencephalographic recording on adult zebrafish using custom-designed electrodes and perfusion system. First, we exploited demonstration of long-term recording on pentylenetetrazole-induced seizure models, and the results were quantified. Second, we studied skin–electrode impedance, which is crucial to the quality of signals. Then, seizure propagations and gender differences in adult zebrafish were exhibited for the first time. Our results provide a new pathway for future neuroscience research using zebrafish by overcoming the challenges for aquatic organisms such as precision, serviceability, and continuous water seepage.

Recording nerve signals in canine sciatic nerves with a flexible penetrating microelectrode array

OBJECTIVE Previously, we presented the fabrication and characterization of a flexible penetrating microelectrode array (FPMA) as a neural interface device. In the present study, we aim to prove the feasibility of the developed FPMA as a chronic intrafascicular recording tool for peripheral applications. APPROACH For recording from the peripheral nerves of medium-sized animals, the FPMA was integrated with an interconnection cable and other parts that were designed to fit canine sciatic nerves. The uniformity of tip exposure and in vitro electrochemical properties of the electrodes were characterized. The capability of the device to acquire in vivo electrophysiological signals was evaluated by implanting the FPMA assembly in canine sciatic nerves acutely as well as chronically for 4 weeks. We also examined the histology of implanted tissues to evaluate the damage caused by the device. MAIN RESULTS Throughout recording sessions, we observed successful multi-channel recordings (up to 73% of viable electrode channels) of evoked afferent and spontaneous nerve unit spikes with high signal quality (SNR  >  4.9). Also, minor influences of the device implantation on the morphology of nerve tissues were found. SIGNIFICANCE The presented results demonstrate the viability of the developed FPMA device in the peripheral nerves of medium-sized animals, thereby bringing us a step closer to human applications. Furthermore, the obtained data provide a driving force toward a further study for device improvements to be used as a bidirectional neural interface in humans.

Development of integrated flexible penetrating microelectrode array with interconnection cable for use in various nervous systems

We developed a flexible and penetrating microelecrode arrays (FPMA) and interconnection cables for neuronal signal recording. For successful implantation of FPMAs into various locations of the nervous system, FPMAs in different dimensions were fabricated using established fabrication process protocols in previous studies. Also, the interconnection cables based on parylene-C were designed and fabricated for implantations in cortex and peripheral nerves. In order to investigate the feasibility of the proposed integrated FPMA with the interconnection cable as a neural recording device, we implanted it onto the cortex of rats and the sciatic nerve of a dog for acute signal recording. Using the developed FPMA integrated with interconnection cable, acute neural signals were successfully recorded from both cortex and sciatic nerves.

Finite element analysis of the biomechanical effect of coflex™ on the lumbar spine.

Objective The biomechanical properties of the Coflex™ (Paradigm Spine, NY, USA), a device designed to provide dynamic stabilization without lumbar fusion, have not been clearly defined. The purpose of this study was to determine the efficacy and biomechanical effect of Coflex™ using finite element model (FEM). Methods A 3D geometric model of the L3-L5 was created by integrating computerized tomography (CT) images. Based on the geometric model, a 3D FEM was created and the Coflex™ model was incorporated into the base model. Mechanical load dependent on the postural changes and boundary conditions, were imposed to simulate various 3D physiological states. The simulation analysis included stress and strain distributions, intervertebral disc deformation, and the range of motion of the facet joint and lumbar spinous process. Results Coflex™ significantly restrained displacement in extension, lateral bending and compression of joint between the L4-5 as one in the experimental group was observed -1.3% of flexion, -24.5% of extension, -44.5% of lateral bending and -37.2%. The average intradiscal pressure of the L4-5 decreased by 63% and the average facet contract force of the L4-5 decreased markedly by 34% in the experimental group. A load of 120 MPa from extension was observed at the base of spinous process in the experimental group. Conclusion The Coflex™ can be safely used for achieving functional dynamic stabilization of the lumbar vertebral column while preserving the intactness of the other components. However, the fatigue fracture of the L4 spinous process should be carefully monitored.

Simplified adaptor for stereotactic surgery in non-human primates

BACKGROUND It is challenging for researchers performing stereotactic procedures to transition from small animals to non-human primate (NHP) experiments. The NHP stereotactic atlas is based on ear-bar zero (EBZ), which is an anatomical reference frame that is not visible during surgery. Most current NHP stereotactic systems require high-cost MRI or CT imaging and complex computer processing to determine the stereotactic coordinates, limiting the procedure to those with significant expertise. NEW METHOD We have designed a simplified adaptor consisting of a circular arc for coronal tilt, a carrier for electrodes or cannulas, and an anchor to attach the adaptor to a conventional stereotactic frame. Our adaptor allows easy identification of the EBZ with the help of an anchor notch, and provides digital distance sensors without the need for imaging data or computer processing. Our system enables the use of trajectories that avoid injury to important structures and vessels. RESULTS We tested the accuracy of our system using simulated targeting with phantoms, and demonstrated sub-millimeter accuracy. Infusion of methylene blue also showed satisfactory staining in target structures deep in the brain. COMPARISON WITH EXISTING METHODS This system does not require high-cost imaging and extra training to determine EBZ. Once EBZ is set automatically by the system itself, targeting is similar to that in small animal stereotactic procedure. CONCLUSION Our simple adaptor will aid researchers who plan to conduct experiments involving stereotactic surgery in NHPs.

A 3D-Printed Sensor for Monitoring Biosignals in Small Animals

Although additive manufacturing technologies, also known as 3D printing, were first introduced in the 1980s, they have recently gained remarkable popularity owing to decreased costs. 3D printing has already emerged as a viable technology in many industries; in particular, it is a good replacement for microfabrication technology. Microfabrication technology usually requires expensive clean room equipment and skilled engineers; however, 3D printing can reduce both cost and time dramatically. Although 3D printing technology has started to emerge into microfabrication manufacturing and medical applications, it is typically limited to creating mechanical structures such as hip prosthesis or dental implants. There have been increased interests in wearable devices and the critical part of such wearable devices is the sensing part to detect biosignals noninvasively. In this paper, we have built a 3D-printed sensor that can measure electroencephalogram and electrocardiogram from zebrafish. Despite measuring biosignals noninvasively from zebrafish has been known to be difficult due to that it is an underwater creature, we were able to successfully obtain electrophysiological information using the 3D-printed sensor. This 3D printing technique can accelerate the development of simple noninvasive sensors using affordable equipment and provide an economical solution to physiologists who are unfamiliar with complicated microfabrication techniques.

Polymer-based interconnection cables to integrate with flexible penetrating microelectrode arrays

There have been various types of interconnection methods for neural interfacing electrodes, such as silicon ribbon cables, wire bonding and polymer-based cables. In this study, interconnection cables were developed for integration with a Flexible Penetrating Microelectrode Array (FPMA) that was previously developed for neural signal recording or stimulation. Polyimide and parylene C were selected as base materials for the interconnection cables as both materials can preserve the flexibility of the FPMA better than other interconnection methods such as silicon ribbon cable or wire bonding. We conducted durability tests to determine if the interconnection cables were suitable for in-vivo implantation, by long-term soaking of the cables in phosphate buffered saline solution. We measured the changes in impedance over time, and equivalent circuit models were used to analyze the electrochemical phenomena on the surface of the cables. Lastly, we implanted the cable-integrated electrodes device onto rabbit’s sciatic nerve and recorded neural signals to prove the feasibility of the developed FPMA integration system.