Morphological assessment of early axonal regeneration in end-to-side nerve coaptation models

Abstract

Histological changes were observed in peripheral nerves following end-to-side nerve coaptation to determine the effects of perineurial opening and deliberate donor nerve injury during surgery. Twenty rats were randomised into four groups as follows: group 1, end-to-side nerve coaptation without perineurial opening; group 2, end-to-side nerve coaptation with simple perineurial opening; group 3, end-to-side nerve coaptation with partial crush injury after perineurial opening; group 4, end-to-side nerve coaptation with partial neurotomy after perineurial opening. Seven days after coaptation of the musculocutaneous (recipient) nerve to the ulnar (donor) nerve, the nerves were immunohistochemically analysed using antibodies against neurofilament-H (RT97) and phosphorylated GAP-43 (p-GAP-43). The former labels all axons, including regenerating axons and degenerated axonal debris, while the latter only labels regenerating axons. Results demonstrated no regenerating nerves in the recipient nerve of group 1. In group 2, because nerve herniation from the perineurial opening partially injured donor nerve fibres, some regenerating axons extended proximally and distally along the partially injured fibres in the donor nerve; some of these regenerating axons also extended into the recipient nerve via the perineurial opening. In groups 3 and 4, thin regenerating axons were more prominent in recipient and donor nerves compared with group 2. Statistical evaluation revealed increased efficacy of perineurial opening and deliberate donor nerve injury in end-to-side nerve coaptation, suggesting that partial nerve fibre herniation with partial axonotmesis or neurotomesis was important for effective axonal regeneration in end-to-side nerve coaptation.

Key Words::

• End-to-side nerve coaptation
• GAP-43
• nerve injury
• nerve regeneration
• perineurial window
• peripheral nerve
• RT97

Acknowledgements

We thank Dr Motohiro Nozumi and Dr Kosei Takeuchi (Division of Molecular and Cellular Biology, Niigata University Graduate School of Medical and Dental Sciences) for their contributions to characterisation of p-GAP-43 antibody. We also thank Professor K. Akazawa (Department of Medical Informatics, Niigata University Medical and Dental Hospital) for professional statistical analysis advice, and members of the Division of Microscopic Anatomy, Niigata University Graduate School of Medical and Dental Sciences for generous support throughout this study.