Optimal electrical stimulation boosts stem cell therapy in nerve regeneration methods
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Materials and methods
Our electrostimulation study was performed with a built-in-house platform that generates a vertical electric field with electrodes placed in a top-bottom configuration (illustrated in ). The vertical configuration contrasts with conventional horizontal electrical potential gradients [ ] by its ability to stimulate a larger area of cells and separate the effects of galvanic migration resulting from cell differentiation. The top electrodes (99.999% pure gold wires, Surepure, Chemetals, Florham Park, NJ, USA) are connected to a printed circuit board, and the bottom electrodes are made by depositing chromium and gold onto glass via e-beam vapor deposition. The gold nature of the two electrodes prevents electrochemical reactions with the sample solution during stimulation. The deposition process for the bottom electrode involved cleaning the glass with Piranha solution for 10 min, rinsing...
Intrafascicular ES of axotomized and repaired nerves
In the stimulation groups, after nerve repair, 2 modified intrafascicular electrodes were implanted in both sides of the sciatic nerve stump 0.5 cm from the suture site, as we previously described [ - ]. The insulated electrodes were led to a stimulator (Grass S48 Square Pulse stimulator, Astro-Med Inc, West Warwick, RI). ES was applied between the cathode placed at the distal stump and the anode at the proximal stump. The nerve and their regenerating axons were subject to 1-h continuous ES immediately after repair of the nerve, with supramaximal pulses (100 µsec; 3 V) delivered in a continuous 20 Hz train based on the in vitro study. The intrafascicular electrodes were removed after ES and the incision was closed in layers. The illustration of animal experimental design and procedures was shown in.
Behavioral assessment
We have described the methodology of CatWalk (Noldus Information Technology, The Netherlands) before [,, ]. Briefly, one week prior to surgery, animals were trained daily on a CatWalk runway until they were able to make consecutive uninterrupted runs. Animals were placed on the right side of a runway and motivated to cross the runway toward the left end. Six compliant trials were recorded by a computer running Catwalk XT 10.5 Software (Noldus) for each animal per time point, and five animals for each group. All trials marked by the software as compliant were reviewed manually: if the animal stopped or turned in mid-run the trials were excluded in the analysis. Thus a minimum of three validated runs were available each time point per animal. The stand time and duty cycle parameters, which show the time spent per step and percentage of time per step cycle spent bearing weight for each...
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Gait recovery and locomotor function
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Evidence quotes (3)
Our electrostimulation study was performed with a built-in-house platform that generates a vertical electric field with electrodes placed in a top-bottom configuration (illustrated in ). The vertical configuration contrasts with conventional horizontal electrical potential gradients [ ] by its ability to stimulate a larger area of cells and separate the effects of galvanic migration resulting from cell differentiation. The top electrodes (99.999% pure gold wires, Surepure, Chemetals, Florham Park, NJ, USA) are connected to a printed circuit board, and the bottom electrodes are made by depositing chromium and gold onto glass via e-beam vapor deposition. The gold nature of the two electrodes prevents electrochemical reactions with the sample solution during stimulation. The deposition process for the bottom electrode involved cleaning the glass with Piranha solution for 10 min, rinsing with deionized water, and then air-drying. Then, via patterned mask, an electrode array was created by vapor depositing a 3-nm chromium layer and a 12-nm gold layer onto the glass surface. This design provides a stable coating on the glass and leaves enough optical transparency for light transmissio...
In the stimulation groups, after nerve repair, 2 modified intrafascicular electrodes were implanted in both sides of the sciatic nerve stump 0.5 cm from the suture site, as we previously described [ - ]. The insulated electrodes were led to a stimulator (Grass S48 Square Pulse stimulator, Astro-Med Inc, West Warwick, RI). ES was applied between the cathode placed at the distal stump and the anode at the proximal stump. The nerve and their regenerating axons were subject to 1-h continuous ES immediately after repair of the nerve, with supramaximal pulses (100 µsec; 3 V) delivered in a continuous 20 Hz train based on the in vitro study. The intrafascicular electrodes were removed after ES and the incision was closed in layers. The illustration of animal experimental design and procedures was shown in.
We have described the methodology of CatWalk (Noldus Information Technology, The Netherlands) before [,, ]. Briefly, one week prior to surgery, animals were trained daily on a CatWalk runway until they were able to make consecutive uninterrupted runs. Animals were placed on the right side of a runway and motivated to cross the runway toward the left end. Six compliant trials were recorded by a computer running Catwalk XT 10.5 Software (Noldus) for each animal per time point, and five animals for each group. All trials marked by the software as compliant were reviewed manually: if the animal stopped or turned in mid-run the trials were excluded in the analysis. Thus a minimum of three validated runs were available each time point per animal. The stand time and duty cycle parameters, which show the time spent per step and percentage of time per step cycle spent bearing weight for each paw respectively, were collected and blindly assessed. Animals performed runs at the 2, 6, and 12 week time points. In order to prevent the confounding effects of walking speed and animal weight, the non-impaired contralateral hind paws were used for control comparison, and results are reported as...
Machine-readable layer
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