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Axonal Regrowth of Olfactory Sensory Neurons In Vitro.

Rebecca Sipione1, Nicolas Liaudet2, Francis Rousset1

  • 1The Inner Ear and Olfaction Lab, Department of Pathology and Immunology, Faculty of Medicine, University of Geneva, Rue Michel Servet 1, CH-1211 Geneva, Switzerland.

International Journal of Molecular Sciences
|August 26, 2023
PubMed
Summary

Researchers developed an automated tool to measure olfactory neuron regrowth. Fibronectin, collagen IV, FGF2, and NGF significantly enhanced axonal length in a mouse model, paving the way for new treatments for olfactory loss.

Keywords:
anosmiaaxon tracingaxonal regenerationnerve growth factorsolfactory neuronstraumatic brain injury

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Area of Science:

  • Neuroscience
  • Regenerative Medicine
  • Biomaterials

Background:

  • Traumatic brain injury often causes permanent olfactory loss due to olfactory axon shearing at the cribriform plate.
  • Scar tissue impedes axonal regrowth, and current treatments are lacking.
  • Developing effective neuroregeneration therapies requires optimal conditions and efficient assessment methods.

Purpose of the Study:

  • To create an automated tool for quantifying axonal length in olfactory sensory neurons.
  • To identify optimal extracellular matrix proteins and growth factors for enhancing olfactory neuron regeneration in vitro.
  • To establish a mouse organotypic 2D model for studying olfactory neuroregeneration.

Main Methods:

  • Cultured olfactory epithelium (OE) from C57BL/6 mice on coverslips coated with Fibronectin, Collagen IV, or Laminin.
  • Supplemented cultures with various growth factors: FGF2, BDNF, GDNF, NGF, RA, TGFβ, or none.
  • Utilized a newly developed automated high-throughput tool to measure axonal length and compared it to standard methods.

Main Results:

  • Fibronectin and Collagen IV coatings significantly enhanced total axonal length (p=0.001).
  • FGF2 alone, or FGF2 followed by FGF2/NGF combination, maximally promoted axonal regrowth (p<0.0001).
  • The automated tool reduced analysis time from 22 to 3 minutes per specimen compared to Neuron J.

Conclusions:

  • Robust in vitro regeneration of murine olfactory neurons is achievable and controllable.
  • Specific extracellular matrix proteins (Fibronectin, Collagen IV) and growth factors (FGF2, NGF) optimize neuroregeneration.
  • The automated quantification tool significantly improves efficiency for assessing axonal length, advancing therapeutic development.