What is it about?
We explain in this review how the wound-scratch assay, a simple laboratory test, is used to study wound healing. In this test, researchers create a small gap in a layer of cells and observe how quickly the cells move to close it. This helps scientists investigate cell migration and screen potential treatments that may improve wound healing. We review the different ways this test is performed, from simple manual methods to advanced automated systems, and discusses how these improvements have made the test more accurate and reproducible. However, it also highlights an important limitation: the wound-scratch assay only measures cell movement on a flat surface and cannot fully reproduce the complex healing process that occurs in the human body. Real wound healing also involves inflammation, immune cells, blood vessel formation, collagen production, and interactions with surrounding tissues, which are largely missing from this laboratory model. We conclude that the wound-scratch assay remains an excellent first-step research tool, but its results should be interpreted with caution and combined with more advanced laboratory models to better predict how treatments will perform in patients.
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Why is it important?
This review is important because the wound-scratch assay is one of the most widely used laboratory methods for studying wound healing and testing new treatments before they are evaluated in animals or humans. It is simple, affordable, and allows researchers to quickly identify promising compounds that may improve wound repair. At the same time, the review highlights that this test has important limitations. Because it only measures how cells move across a flat surface, it cannot fully represent the complex biological processes involved in real wound healing, such as inflammation, immune responses, blood vessel formation, collagen remodelling, and interactions between different cell types. As a result, some treatments that work well in the body may appear less effective in this laboratory test. By explaining both the strengths and weaknesses of the wound-scratch assay, the review helps researchers choose the right experimental models, improve the reliability of their findings, and avoid misleading conclusions. It also encourages combining the scratch assay with more advanced laboratory models to increase the chances of developing effective therapies for patients with difficult-to-heal wounds.
Perspectives
We suggest that future wound-healing research should move beyond relying on the wound-scratch assay alone. While it will remain a valuable first-step laboratory test, combining it with more advanced models that better mimic human skin will provide more reliable and clinically relevant results. Future studies should incorporate three-dimensional (3D) tissue models, co-culture systems, and technologies that include immune cells, blood vessels, and the extracellular matrix to better reflect how wounds heal in the human body. We also highlight the potential of automated imaging, artificial intelligence, and machine learning to improve data analysis, reduce variability, and help predict which treatments are most likely to succeed in patients. By integrating simple laboratory assays with more advanced experimental models and modern analytical tools, researchers can improve the development and evaluation of new therapies, ultimately leading to more effective treatments for patients with acute and chronic wounds.
Dr Zeinab Ghasemishahrestani
University of Cape Town
Read the Original
This page is a summary of: The In Vitro Wound‐Scratch Assay: Applications, Technical Advances, and Limitations in Wound Healing Research, International Wound Journal, June 2026, Wiley,
DOI: 10.1111/iwj.70964.
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