Peripheral Nerve Injury and Repair Model
Objective: Evaluate the effects of optimized electrical stimulation on human neural crest stem cell (NCSC) transplantation for peripheral nerve regeneration following critical-sized sciatic nerve injury and repair
Gather these items before starting the experiment. Check off items as you prepare.
Equipment2
Materials1
Not specified • Not specified • Not specified • Not specified
Software1
Not specified • Not specified
1 item available from ConductScience
Estimated: $2,610.00
As an Amazon Associate, we earn from qualifying purchases. Product links help support this free resource.
Protocol Steps
In vitro electrical stimulation optimization
Investigated effects of different stimulating pulse frequency, duration and field direction on human neural crest stem cell differentiation using the 96-well vertical ES platform
Note: Optimal conditions identified: 20 Hz, 100 microseconds pulse at a potential gradient of 200 mV/mm applied to cathodes
View evidence from paper
“We observed dendritic morphology with enhanced neuronal differentiation for NCSCs cultured on cathodes subject to 20 Hz, 100μs pulse at a potential gradient of 200 mV/mm”
Generation of critical-sized sciatic nerve injury
Created 15 mm critical-sized sciatic nerve injuries in sixty athymic nude rats
Note: Injuries were followed by surgical repair
View evidence from paper
“15 mm critical-sized sciatic nerve injuries were generated with subsequent surgical repair in sixty athymic nude rats”
Surgical repair and group assignment
Injured animals were randomly assigned into five groups (N = 12 per group): blank control, ES, NCSC, NCSC + ES, and autologous nerve graft
Note: Random assignment to ensure unbiased group distribution
View evidence from paper
“Injured animals were randomly assigned into five groups (N = 12 per group): blank control, ES, NCSC, NCSC + ES, and autologous nerve graft”
Application of optimized electrical stimulation
Optimized ES was applied immediately after surgical repair for 1 hour in ES and NCSC + ES groups
Note: Applied immediately post-surgery using optimized parameters (20 Hz, 100 microseconds pulse, 200 mV/mm gradient)
View evidence from paper
“Optimized ES was applied immediately after surgical repair for 1 h in ES and NCSC + ES groups”
Behavioral assessment - CatWalk gait analysis
Gait analysis performed to assess locomotor recovery and nerve regeneration
Note: Conducted at 6 or 12 weeks post-surgery (N = 6 per group per timepoint)
View evidence from paper
“Recovery was assessed by behavioral (CatWalk gait analysis), wet muscle-mass, histomorphometric, and immunohistochemical analyses at either 6 or 12 weeks after surgery”
Wet muscle-mass measurement
Gastrocnemius muscle wet mass measurements to assess muscle recovery and innervation
Note: Conducted at 6 or 12 weeks post-surgery (N = 6 per group per timepoint)
View evidence from paper
“Gastrocnemius muscle wet mass measurements in ES + NCSC group were comparable to autologous nerve transplantation”
Histomorphometric analysis
Quantitative analysis of nerve tissue morphology and regeneration
Note: Conducted at 6 or 12 weeks post-surgery (N = 6 per group per timepoint)
View evidence from paper
“Quantitative histomorphometric analysis and catwalk gait analysis showed similar improvements by ES on NCSCs”
Immunohistochemical analysis
Assessment of viable NCSCs and Schwann cell differentiation in regenerated nerve tissue
Note: Conducted at 6 or 12 weeks post-surgery (N = 6 per group per timepoint); evaluated NCSC viability and SC differentiation
View evidence from paper
“A higher number of viable NCSCs was shown via immunochemical analysis, with higher Schwann cell (SC) differentiation in the NCSC + ES group compared to the NCSC group”