Background: Donor nerve axonal count over 900 in two-stage reconstructions using cross-facial nerve grafts is possibly associated with improved outcomes in facial reanimation. Facial nerve axonal analysis was performed to determine the ideal location for optimizing axonal load. Correlation of axonal number, branch diameter, and age was also assessed. Methods: Twenty-eight fresh unpreserved cadaveric hemifaces were dissected exposing the extracranial facial nerve branches. Axonal counts at 2-cm intervals from the pes anserinus along branches inserting into the zygomaticus major muscle were taken, noting position relative to the zygomatic arch, posterior ramus border, lateral border of the zygomaticus muscle, and anterior parotid gland border. Nerves were fixed, sectioned, and stained with SMI-31 antineurofilament stain for digital axonal analysis. Results: All specimens had one or more intraparotid zygomatic branches with over 900 axons, and 96 percent had an extraparotid branch with over 900 axons. The likelihood that a zygomatic branch would have over 900 axons at its last intraparotid point (mean, 6 mm posterior to the parotid border) was 92 percent (range, 67 to 100 percent) in contrast to 61 percent (range, 25 to 100 percent) when sampled at the first extraparotid point (mean, 14 mm anterior to the parotid border). Nerve cross-sectional area was positively correlated to its axonal count (R° = 78 percent; p < 0.0001), with nerve diameter over 0.6 mm predicting over 900 axons. Age did not correlate with axonal counts. Conclusions: Branches with adequate axonal load were found in all specimens. The likelihood of adequate branch selection improved from 61 percent to 92 percent with short retrograde intraparotid dissection. Nerve diameter correlated with axonal load.
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