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Related Experiment Video

Updated: Jan 10, 2026

Scratch Migration Assay and Dorsal Skinfold Chamber for In Vitro and In Vivo Analysis of Wound Healing
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A novel method for evaluating and visualizing scratch wound healing assays using level-set and image sector analysis.

Markéta Vašinková1, Michal Krumnikl1, Arootin Gharibian1

  • 1Department of Computer Science, FEECS, VSB - Technical University of Ostrava, 17. listopadu 2172/15, Ostrava 70800, Czech Republic.

PNAS Nexus
|November 24, 2025
PubMed
Summary

A new sector-based analysis framework reveals how cell migration patterns influence wound healing. This method enhances understanding of tissue regeneration and drug effects by spatially resolving cell movement, showing treatment-dependent healing contributions.

Keywords:
collective cell migrationimage processinglevel-setsscratch wound healing assaytime-lapse microscopy

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Area of Science:

  • Cell Biology
  • Biophysics
  • Regenerative Medicine

Background:

  • Scratch wound healing assays are crucial for studying collective cell migration in tissue regeneration and drug development.
  • Traditional analysis methods for these assays often lack spatial resolution, limiting insights into complex cell migration behaviors.

Purpose of the Study:

  • To develop and validate a novel sector-based analytical framework for spatially resolved characterization of cell migration in scratch wound assays.
  • To investigate the influence of local cellular activity and chemotactic stimulation on wound closure dynamics.

Main Methods:

  • A sector-based analytical framework was implemented to divide microscopy images into spatial regions for migration analysis.
  • A level-set segmentation algorithm was employed for robust wound edge detection, addressing challenges in bright-field microscopy.
  • Cell velocity, trajectory, and motility were analyzed in relation to distance from the wound boundary, including assessment of CXCL10-stimulated migration.

Main Results:

  • Cell velocity and trajectory were found to vary significantly with distance from the wound boundary.
  • A novel metric, sector-boundary distance, identified regions of faster, non-radial cell migration.
  • Chemokine (CXCL10) treatment significantly increased the proportion of highly motile cells associated with wound closure, even in distant regions, while directionality played a minor role.

Conclusions:

  • The developed sector-based framework effectively bridges global and local analyses of cell migration in wound healing.
  • Local cellular activity, particularly enhanced motility upon chemotactic stimulation, significantly contributes to wound closure in a treatment-dependent manner.
  • The open-source software provides an accessible tool for interactive analysis of microscopy data, advancing cell migration research.