Using the Neuromuscular Junction Cellular Response to Predict Functional Recovery after Nerve Grafting in a Mouse Experimental Model.

BACKGROUND

Functional recovery following peripheral nerve injury worsens with increasing time of denervation before repair. Denervated muscle undergoes progressive atrophy that limits the extent to which motor endplates can be reinnervated. The aims of this study were to assess nerve injuries reconstructed at different time points and to identify various neural and muscle-based markers to predict functional outcome, including an in-depth look at the neuromuscular junction (NMJ).

METHODS

Adult wild-type C57BL/6J mice underwent surgery on the sciatic nerve and were divided into 5 groups: (1) nerve cut and repaired, (2) acute (nerve cut and immediately repaired with a 1-cm autograft), (3) subacute (nerve grafted 2 weeks after injury), (4) delayed (nerve grafted 4 weeks after injury), (5) nerve cut and capped. Functional recovery was measured by treadmill and electrodiagnostic tests. Nerves were harvested for histologic evaluation, and leg muscles, for histologic evaluation and NMJ immunofluorescent staining of motor endplate innervation and terminal Schwann cells (tSCs).

RESULTS

The delayed graft group performed worst in nearly all parameters. The subacute graft group shared more similarities with the acute group, especially the tSC response (subacute, 48%; acute, 51%) and motor endplate innervation pattern (subacute, 75%; acute, 72%). The only parameters to elucidate differences were muscle weight and motor endplate fragmentation. Traditional axon count failed to capture differences among the 3 groups.

CONCLUSION

The tSC activity and NMJ innervation pattern can be used as predictive markers of functional recovery that capture differences among acute, subacute, and delayed nerve injuries.

CLINICAL RELEVANCE STATEMENT: The tSC activity at the NMJ displayed by immunohistochemical methods can accurately reflect target muscle innervation over time. This can be used as a predictor of reinnervation when timing of nerve reconstruction is delayed, which can be a critically needed tool in clinical scenarios.