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Quantifying innervation facilitated by deep learning in wound healing.

Abijeet Singh Mehta1,2, Sam Teymoori3, Cynthia Recendez4,5

  • 1Department of Dermatology, University of California, Davis, CA, 95616, USA. abijeet.mehta@northwestern.edu.

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Peripheral nerves (PNs) are crucial for wound healing. This study developed an automated image analysis method using deep neural networks to quantify skin re-innervation, revealing a significant increase in nerve density over time.

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Area of Science:

  • Neuroscience
  • Dermatology
  • Biomedical Engineering

Background:

  • Peripheral nerves (PNs) play a key role in wound healing.
  • Current methods for quantifying skin innervation are complex, labor-intensive, and prone to bias.
  • Immunohistochemistry (IHC) images often contain noise that complicates accurate quantification.

Purpose of the Study:

  • To develop and validate an automated image analysis method for quantifying skin innervation during wound healing.
  • To establish a quantitative time course of re-innervation in a mouse wound healing model.
  • To investigate the relationship between re-innervation and re-epithelization.

Main Methods:

  • Utilized a Denoising Convolutional Neural Network (DnCNN) to reduce noise in IHC images.
  • Employed an automated image analysis tool (Matlab-assisted) for precise quantification of nerve fibers.
  • Generated 8mm wounds in wild-type mice, collecting samples on days 3, 7, 10, and 15 post-wounding.
  • Stained tissue sections with protein-gene-product 9.5 (PGP 9.5) antibody, a pan-neuronal marker.

Main Results:

  • Negligible nerve fibers were observed on days 3 and 7 post-wounding, primarily at wound boundaries.
  • A slight increase in nerve fiber density was noted on day 10, with a significant increase by day 15.
  • A strong positive correlation (R² = 0.926) was found between nerve fiber density and the extent of re-epithelization.

Conclusions:

  • The developed automated method effectively quantifies skin innervation during wound healing.
  • Re-innervation significantly increases in the later stages of wound healing (days 10-15).
  • Re-innervation is positively associated with re-epithelization, highlighting its importance in the healing process.