doi: 10.3967/bes2023.001
Comparison of the Nerve Regeneration Capacity and Characteristics between Sciatic Nerve Crush and Transection Injury Models in Rats
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Abstract:
Objective To provide useful information for selecting the most appropriate peripheral nerve injury model for different research purposes in nerve injury and repair studies, and to compare nerve regeneration capacity and characteristics between them. Methods Sixty adult SD rats were randomly divided into two groups and underwent crush injury alone (group A, n = 30) or transection injury followed by surgical repair (group B, n = 30) of the right hind paw. Each group was subjected to the CatWalk test, gastrocnemius muscle evaluation, pain threshold measurement, electrophysiological examination, retrograde neuronal labeling, and quantification of nerve regeneration before and 7, 14, 21, and 28 days after injury. Results Gait analysis showed that the recovery speed in group A was significantly faster than that in group B at 14 days. At 21 days, the compound muscle action potential of the gastrocnemius muscle in group A was significantly higher than that in group B, and the number of labeled motor neurons in group B was lower than that in group A. The number of new myelin sheaths and the g-ratio were higher in group A than in group B. There was a 7-day time difference in the regeneration rate between the two injury groups. Conclusion The regeneration of nerve fibers was rapid after crush nerve injury, whereas the transection injury was relatively slow, which provides some ideas for the selection of clinical research models. -
Key words:
- Sciatic nerve injury /
- Degeneration /
- Regeneration /
- Myelination
No conflict of interest to declare.
注释:1) AUTHORS’ CONTRIBUTIONS: 2) CONFLICT OF INTEREST: -
Figure 1. Establishment of the two models of sciatic nerve injury. (A1–A2) Representative images of the method used to establish the crush injury model. (A3) Enlarged view of the section marked in A2. (A4) The sciatic nerve tissue H&E staining of A3 revealed the presence of collagen fibers in the epineurium, magnification = 10x. (B1–B2) Representative images of the method used to establish the transection injury model. (B3) Enlarged view of the section marked in B2. (B4) The sciatic nerve tissue H&E staining of B3 revealed the structural damage and disorder of collagen fibers in the epineurium, magnification = 10x
Figure 2. Evaluation of the magnitude and extent of dynamic and static gait changes before and after sciatic nerve injury in adult rats. (A–C) Representative diagram of rat gait under fluorescence photography of the track. (D) The sciatic nerve function index at different times; two-way repeated-measures ANOVA: F4,31 = 4.279, P < 0.01; day 21, P < 0.01; day 28, P < 0.01. (E) Hind limb stance width; two-way repeated-measures ANOVA: F4,36 = 21.58 P < 0.0001; day 14, P < 0.0001; day 28, P < 0.05. (F) Stance duration ratio between the ipsilateral and contralateral sides; two-way repeated-measures ANOVA: F4,33 = 2.893, P < 0.05; day 21, P < 0.05. (G) Swing duration ratio between the ipsilateral and contralateral sides; two-way repeated-measures ANOVA: F4,33 = 2.081, P = 0.1. *Represents the post hoc test comparing the two groups at the same time point.
Figure 3. Evaluation of the pressure threshold for the hind paw. (A) Calibrated forceps and measurement technique. (B) Pressure threshold trend over time; two-way repeated-measures ANOVA: F4,61 = 11.01, P < 0.0001; day 7, P < 0.01; day 14, P < 0.001; day 21, P < 0.05; day 28, P < 0.0001. *Represents the post hoc test comparing the two groups at the same time point.
Figure 4. Morphology of the gastrocnemius. (A1–A4) Morphology and H&E staining of the gastrocnemius muscle 7, 14, 21 and 28 days after sciatic nerve injury in group A, bar = 20 µm. (B1–B4) Morphology and H&E staining of the gastrocnemius muscle 7, 14, 21 and 28 days after sciatic nerve injury in group B, bar = 20 µm. (C) Normal morphology of the gastrocnemius. (D) Wet weight ratio between the ipsilateral and contralateral muscles; two-way repeated-measures ANOVA: F4,50 = 24.73, P < 0.0001; day 21, P < 0.01; day 28, P < 0.0001. *Represents the post hoc test comparing the two groups at the same time point.
Figure 5. Representative quantitative results of the cMAP latency time and amplitudes. (A1–A4) Schematic diagram of amplitude and latency time at 7, 14, 21, and 28 days after sciatic nerve injury in group A (X-axis units: time course 20 ms, Y-axis unit: amplitude 5 mV), where the cMAP latency time is the position of number 1, the amplitude is the vertical distance between peaks (number 2) and troughs (number 3), and the other 3 and 5 represent the baseline position and the waveform end position. (B1–B4) Schematic diagram of the amplitude and latency time at 7, 14, 21, and 28 days after sciatic nerve injury in group B (the number marks are the same as in group A). (C) Schematic diagram of amplitude and latency time before sciatic nerve injury (the number marks are the same as in group A). (D) Quantitative results of the latency time changes in the two groups at different times. Two-way repeated-measures ANOVA: F4,39 = 4.639, P < 0.01; day 14, P < 0.05; day 28, P < 0.05; (E) Quantitative results of cMAP amplitudes in the two injury groups at different time points; two-way repeated-measures ANOVA: F4,33 = 7.057, P < 0.001; day 21, P < 0.05; day 28, P < 0.0001. *Represents the post hoc test comparing of the two groups at the same time point.
Figure 6. Quantitative results following CTB-Alexa 555 labeling of neurons in the right spinal cord. (A1–A4) Labeled neurons in group A 7, 14, 21, and 28 days after sciatic nerve injury, magnification = 10x. (B1–B4) Labeled neurons in group B 7, 14, 21, and 28 days after sciatic nerve injury, magnification = 10x. (C) CTB-Alexa 555-labeled neurons in the spinal cord of a rat before sciatic nerve injury, magnification = 10x. (D) Quantitative results of the two groups; two-way repeated-measures ANOVA: F4,22 = 17.54, P < 0.0001; day 21, P < 0.0001; day 28, P < 0.0001. *Represents the post hoc test comparing of the two groups at the same time point.
Figure 7. Representative photomicrographs showing myelinated axons in thionin-stained semithin sections of the sciatic nerve at different times. The black triangles indicate newly formed myelin sheaths, and the stars represent mature myelin sheaths. (A1–A4) Myelinated axons in group A 7, 14, 21, and 28 days after sciatic nerve injury, bar = 60 µm. (B1–B4) Myelinated axons in group B 7, 14, 21, and 28 days after sciatic nerve injury, bar = 60 µm. (C) Photomicrographs of myelinated axons before sciatic nerve injury, bar = 60 µm. (D) Numbers of myelinated axons for the two groups; two-way repeated-measures ANOVA: F4,20 = 49.69, P < 0.0001; day 14, P < 0.05; day 21, P < 0.05; day 28, P < 0.001. *Represents the post hoc test comparing of the two groups at the same time point.
Figure 8. Representative electron micrographs showing myelinated axons in ultrathin sections of the sciatic nerve at different times. The white triangles indicate newly formed myelin sheaths, the red triangles indicate myelin sheaths undergoing disintegration, the red pentagrams represent Schwann cell nuclei, and the black triangles represent mature myelin sheaths. (A1–A4) Electron micrographs of myelinated axons in group A 7, 14, 21, and 28 days after sciatic nerve injury, bar = 5 µm. (B1–B4) Electron micrographs of myelinated axons in group B 7, 14, 21, and 28 days after sciatic nerve injury, bar = 5 µm. (C) Structure and morphology of myelinated axons before sciatic nerve injury, bar = 5 µm. (D) Quantitative results for the g-ratio (ratio between fiber diameter and axon diameter); two-way ANOVA: F4,26 = 3.583, P < 0.05, day 7 P < 0.05.
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