Fluorescent Reporter Mice for Nerve Guidance Conduit Assessment: A High-Throughput in vivo Model

Laryngoscope. 2018 Nov;128(11):E386-E392. doi: 10.1002/lary.27439. Epub 2018 Aug 10.

Abstract

Objective: Use of cell culture and conventional in vivo mammalian models to assess nerve regeneration across guidance conduits is resource-intensive. Herein we describe a high-throughput platform utilizing transgenic mice for stain-free axon visualization paired with rapid cryosection techniques for low-cost screening of novel bioengineered nerve guidance conduit performance.

Methods: Interposition repair of sciatic nerve transection in mice expressing yellow fluorescent protein in peripheral neurons (Thy1.2 YFP-16) was performed with various bioengineered neural conduit compositions using a rapid sutureless entubulation technique under isoflurane anesthesia. Axonal ingrowth was assessed at 3 and 6 weeks using epifluorescent microscopy following cryosectioning.

Results: Mean procedure time (incision-to-closure) was less than 2½ minutes. Direct operational costs of a 3-week experiment was calculated at $21.47 per animal. Tissue processing steps were minimized to aldehyde fixation, cryoprotection and sectioning, and rapid fluorescent dye staining for conduit visualization. Fluorescent microscopy readily resolved robust axonal sprouting at 3 weeks, with clear elucidation of ingrowth-permissive, semipermissive, or restrictive nerve guidance conduit environments.

Conclusion: A rapid and cost-efficient in vivo platform for screening of nerve guidance conduit performance has been described.

Level of evidence: NA. Laryngoscope, E392-E392, 2018.

Keywords: Microscopy; animal; axons; confocal; disease models; fluorescence; guided tissue regeneration; materials testing; mice; nerve guidance conduit; nerve regeneration; neuronal outgrowth; peripheral nerve injuries; prosthesis implantation; sciatic nerve; transgenic.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Axons / physiology
  • Cell Culture Techniques
  • Female
  • Fluorescent Antibody Technique / economics
  • Fluorescent Antibody Technique / methods*
  • Guided Tissue Regeneration / economics
  • Guided Tissue Regeneration / methods*
  • Male
  • Mice
  • Mice, Transgenic
  • Microscopy, Fluorescence / economics
  • Microscopy, Fluorescence / methods*
  • Nerve Regeneration / physiology*
  • Operative Time
  • Sciatic Nerve / injuries*
  • Sciatic Nerve / surgery
  • Tissue Scaffolds*