Efficient and Rapid Histomorphometry of Regenerating Peripheral Nerve.

Area of Science:

• Neuroscience
• Biomedical Engineering
• Surgical Innovation

Background:

• Peripheral nerve surgery aims to restore function by repairing or transferring nerves.
• Accurate assessment of axon counts in operated nerves is crucial for functional outcomes but lacks reliable intraoperative methods.
• Conventional histologic assessment is time-consuming, hindering timely surgical decisions.

Purpose of the Study:

• To develop and validate a high-throughput, rapid protocol for histomorphometry of regenerating peripheral nerves.
• To enable intraoperative assessment of axon counts for improved surgical guidance.
• To advance peripheral nerve surgery techniques and patient outcomes.

Main Methods:

• Utilized commercial fluorescent membrane-specific dyes (e.g., MemBrite) for staining peripheral nerve sections.
• Employed confocal microscopy and commercial segmentation software for axon quantification.
• Compared results with conventional neurofilament immunohistochemistry and applied the protocol in a clinical case.

Main Results:

• The novel rapid staining protocol provided axon quantification results comparable to conventional methods.
• Minimal relative difference (<1%) in axon counts for healthy murine facial nerves.
• A 8.7% relative difference in axon counts for regenerating human nerves.

Conclusions:

• A rapid, high-throughput protocol for quantifying regenerating axons has been established.
• This method offers potential for intraoperative decision-making in peripheral nerve surgery.
• Faster feedback on axon counts can lead to improved clinical outcomes, particularly in cross-facial nerve surgeries.