Two or 10 weeks after cuff implantation, mice were deeply anesthetized with a ketamine, xylazine, and acepromazine cocktail and then transcardially perfused with cold 4% paraformaldehyde in phosphate-buffered saline. Sciatic nerves were dissected from mice implanted with the device or PE cuff or from sham mice and post-fixed in 4% paraformaldehyde overnight. Nerves were embedded in paraffin and longitudinal sections were cut at 6 μm. Slides were stained with a standard hematoxylin and eosin protocol (18, 44). Representative 40× images taken from the center of the nerve were obtained using a Leica DM6 B microscope, and infiltrating immune cells were counted (18). Three mice were analyzed for each condition, and three nerve sections were evaluated from each mouse. Two images were obtained from each nerve section, and the average number of immune cells present in the six images was calculated. The investigator performing the analysis was blinded to the experimental condition.

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