Fabrication and electromechanical characterization of near-field electrospun composite fibers.

We report the use of near-field electrospinning (NFES) as a route to fabricate composite electrodes. Electrodes made of composite fibers of multi-walled carbon nanotubes in polyethylene oxide (PEO) are formed via liquid deposition, with precise control over their configuration. The electromechanical properties of free-standing fibers and fibers deposited on elastic substrates are studied in detail. In particular, we examine the elastic deformation limit of the resulting free-standing fibers and find, similarly to bulk PEO composites, that the plastic deformation onset is below 2% of tensile strain. In comparison, the apparent deformation limit is much improved when the fibers are integrated onto a stretchable, elastic substrate. It is hoped that the NFES fabrication protocol presented here can provide a platform to direct-write polymeric electrodes, and to integrate both stiff and soft electrodes onto a variety of polymeric substrates.

Uniaxial strain of cultured mouse and rat cardiomyocyte strands slows conduction more when its axis is parallel to impulse propagation than when it is perpendicular.

AIM: Cardiac tissue deformation can modify tissue resistance, membrane capacitance and ion currents and hence cause arrhythmogenic slow conduction. Our aim was to investigate whether uniaxial strain causes different changes in conduction velocity (θ) when the principal strain axis is parallel vs perpendicular to impulse propagation. METHODS: Cardiomyocyte strands were cultured on stretchable custom microelectrode arrays, and θ was determined during steady-state pacing. Uniaxial strain (5%) with principal axis parallel (orthodromic) or perpendicular (paradromic) to propagation was applied for 1 minute and controlled by imaging a grid of markers. The results were analysed in terms of cable theory. RESULTS: Both types of strain induced immediate changes of θ upon application and release. In material coordinates, orthodromic strain decreased θ significantly more (P < .001) than paradromic strain (2.2 ± 0.5% vs 1.0 ± 0.2% in n = 8 mouse cardiomyocyte cultures, 2.3 ± 0.4% vs 0.9 ± 0.5% in n = 4 rat cardiomyocyte cultures, respectively). The larger effect of orthodromic strain can be explained by the increase in axial myoplasmic resistance, which is not altered by paradromic strain. Thus, changes in tissue resistance substantially contributed to the changes of θ during strain, in addition to other influences (eg stretch-activated channels). Besides these immediate effects, the application of strain also consistently initiated a slow progressive decrease in θ and a slow recovery of θ upon release. CONCLUSION: Changes in cardiac conduction velocity caused by acute stretch do not only depend on the magnitude of strain but also on its orientation relative to impulse propagation. This dependence is due to different effects on tissue resistance.

In vitro evaluation of gel-encapsulated adipose derived stem cells: Biochemical cues for in vivo peripheral nerve repair.

Adipose-derived stem cells (ASC) are becoming one of the most exploited cells in peripheral nerve repair. They are fast-growing and able to protect neurons from apoptosis; they can reduce post-injury latency and the risk of muscle atrophy. This study evaluates laminin-loaded fibrin gel as an ASC-carrying scaffold for nerve repair. In vitro, ASC retained their proliferative activity but showed significant increase in proliferation rate when encapsulated in gels with low laminin concentrations (i.e., 1 µg/mL). We observed a linear decrease of ASC proliferation rate with increasing laminin concentration from 1 to 100 µg/mL. We next examined the effect of the ASC-carrying fibrin gels on in vitro dorsal root ganglia (DRG) neurite extension, then in vivo sciatic nerve regeneration in adult rats. The ASC-carrying gel was embedded in 15-mm-long, 1.5-mm-diameter polydimethylsiloxane regenerative conduits for in vivo evaluation. At 8-week post implantation, robust regeneration was observed across the long gap. Taken together, these results suggest ASC-carrying gels are a potential path to improve the efficacy of nerve regeneration through artificial guidance conduits and electrode nerve interfaces.

A wireless system with stimulation and recording capabilities for interfacing peripheral nerves in rodents.

Considerable progress has been made in the last decade in implantable bioelectronic neurosystems. Yet most neural implants are used in acute and tethered experimental conditions. Here, we present a preliminary prototype of a multichannel system for simultaneous peripheral nerve stimulation and neural recording. The system comprises miniaturized electronics with a total volume of less then 1.4cm3 including a 3.7V battery which is expected to last for 94 days of standby operation or 18 hours of continuous recording and stimulation. Data read-out and device configuration are wireless. Visceral nerves in rodents are interfaced with compliant extraneural electrodes. The 100×350µm2 electrodes display a low impedance (1.8kn at 1kHz) with a PEDOT:PSS coating. We validated the prototype in acute experiments by applying electrical stimulation to the aortic depressor nerve (ADN), resulting in effective and reproducible decrease in blood pressure and heart rate. The combination of miniaturized electronics and flexible electrodes makes the presented system a versatile platform for future implantable devices interfacing small peripheral nerves and potentially enables new applications in the field of neuroscience.

Use of an implanted sacral nerve stimulator to restore urine voiding in chronically paraplegic dogs.

BACKGROUND: Loss of urinary control after spinal cord injury increases risk of urinary tract disease and is problematical for owners of affected dogs. OBJECTIVES: To design, implant, and test a sacral nerve stimulating device for controlling urine voiding in paraplegic dogs. ANIMALS: Nine pet dogs with severe thoracolumbar spinal cord injury causing paraplegia, loss of hindquarter sensation, and incontinence for more than 3 months. The procedure was offered prospectively to owners of suitable candidates after the irreversibility of the incontinence had been ascertained. METHODS: Open label clinical study. Surgically implantable electrode "books" were designed for insertion and retention of mixed sacral nerves. Sacral nerves were accessed via laminectomy and stimulated to test their ability to elicit detrusor contraction and then inserted into the electrode book, which was attached to a subcutaneously implanted, externally activated receiver. RESULTS: In 8/9 dogs, S2 nerves elicited the largest increases in intravesicular pressure with minimum stimulation and were placed in electrode books. Voiding efficiency was >90% in 8 of the 9 implanted dogs. No important detrimental effects of the procedure were observed. CONCLUSIONS AND CLINICAL IMPORTANCE: This sacral nerve stimulating implant is a simple and apparently effective neuroprosthetic device that restores urine voiding in paraplegic dogs.

Novel use of X-ray micro computed tomography to image rat sciatic nerve and integration into scaffold.

This paper describes how specimens of nervous tissue can be prepared for successful imaging in X-ray Micro Computed Tomography (microCT), and how this method can be used to study the integration of nervous tissue into a polymeric scaffold. The sample preparation involves staining the biological tissue with osmium tetroxide to increase its X-ray attenuation, and a technique for maintaining the specimen in a moist environment during the experiment to prevent drying and shrinkage. Using this method it was possible to observe individual nerve fascicles and their relationship to the 3-D tissue structure. A scaffold supporting a regenerated sciatic nerve was similarly stained to distinguish the nervous tissue from the scaffold, and to observe how the nerve grew through a 2.5 mm long, 100 microm x 100 microm cross-section channel polyimide array. Furthermore, blood vessels could be identified in these images, and it was possible to monitor how a large proximal blood vessel split through the channel scaffold and proceeded down individual channels. This paper explains how microCT is a useful tool both for studying the location and extent of growth into a polymeric scaffold, and for determining whether the regenerated tissue has blood supply.

Assessment of the biocompatibility of photosensitive polyimide for implantable medical device use.

Polyimides have been widely used for biosensor encapsulation and more recently as substrates for neural implants. They have excellent thermal stability, high chemical resistance, and can be prepared as thin, flexible films. Photosensitive polyimides present similar physical properties to polyimides, and have the advantage that they can be photo-lithographically patterned. However, to date little data on their biocompatibility has been reported. Two commercially available polyimides (PI) and one photo-sensitive polyimide (PSPI) were evaluated in vitro using the ISO 10993 standard on biocompatibility. The materials were Dupont Kapton foil HN, HD Microsystem PI2611, and Fujifilm Durimide 7020 (PSPI). PI2611 and Durimide 7020 were spin-coated on silicon wafers, cured at temperatures ranging from 150 to 450 degrees C, and sterilized by autoclave. All materials were evaluated using a scanning electron microscope pre- and postcell culture. Cell viability was determined by an MTS assay. Their mechanical properties and stability during cell culture as a function of time and environment were investigated by nanoindentation. The MTS results show that PSPI is noncytotoxic compared with the negative control of polyethylene and the conventional PIs tested. Fibroblast adhesion, morphology, and spreading were good and better on the PSPI substrate than on the PI2611. Schwann cell appearance was similar on each of the PIs and the PSPI tested. The results suggest that PSPIs may have potential use for biological microsystem and neuroprosthetic applications.