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Updated: Apr 18, 2026
Protocol to Create Chronic Wounds in Diabetic Mice
Published on: September 25, 2019
Generating and reversing chronic wounds in diabetic mice by manipulating wound redox parameters
Sandeep Dhall1, Danh C Do2, Monika Garcia3
1Department of Cell Biology and Neuroscience, University of California, Riverside, 900 University Avenue, Riverside, CA 92521, USA ; Bioengineering Interdepartmental Graduate Program, University of California, Riverside, 900 University Avenue, Riverside, CA 92521, USA.
Abstract:
By 2025, more than 500 M people worldwide will suffer from diabetes; 125 M will develop foot ulcer(s) and 20 M will undergo an amputation, creating a major health problem. Understanding how these wounds become chronic will provide insights to reverse chronicity. We hypothesized that oxidative stress (OS) in wounds is a critical component for generation of chronicity. We used the db/db mouse model of impaired healing and inhibited, at time of injury, two major antioxidant enzymes, catalase and glutathione peroxidase, creating high OS in the wounds. This was necessary and sufficient to trigger wounds to become chronic. The wounds initially contained a polymicrobial community that with time selected for specific biofilm-forming bacteria. To reverse chronicity we treated the wounds with the antioxidants α-tocopherol and N-acetylcysteine and found that OS was highly reduced, biofilms had increased sensitivity to antibiotics, and granulation tissue was formed with proper collagen deposition and remodeling. We show for the first time generation of chronic wounds in which biofilm develops spontaneously, illustrating importance of early and continued redox imbalance coupled with the presence of biofilm in development of wound chronicity. This model will help decipher additional mechanisms and potentially better diagnosis of chronicity and treatment of human chronic wounds.
Insights
Oxidative stress (OS) drives chronic wound development by promoting bacterial biofilms. Antioxidant treatment reversed this, reducing OS and improving healing, offering new therapeutic strategies for diabetic foot ulcers.
Area of Science:
- Wound healing research
- Biomedical engineering
- Microbiology
Background:
- Diabetic foot ulcers represent a significant global health challenge, leading to high rates of amputation.
- Understanding the mechanisms behind chronic wound development is crucial for effective treatment strategies.
Purpose of the Study:
- To investigate the role of oxidative stress (OS) in the chronicity of wounds.
- To explore the interplay between OS, bacterial biofilms, and wound healing.
- To evaluate the efficacy of antioxidant therapy in reversing wound chronicity.
Main Methods:
- Utilized the db/db mouse model to create chronic wounds by inhibiting antioxidant enzymes (catalase, glutathione peroxidase).
- Induced high OS in wounds to observe chronicity development and biofilm formation.
- Treated chronic wounds with antioxidants (α-tocopherol, N-acetylcysteine) to assess reversal of chronicity.
Main Results:
- Successfully generated chronic wounds with spontaneous biofilm development by inducing OS.
- Antioxidant treatment significantly reduced OS, enhanced antibiotic sensitivity of biofilms, and promoted proper tissue regeneration.
- Demonstrated that early and sustained redox imbalance, combined with biofilm presence, is key to wound chronicity.
Conclusions:
- Oxidative stress is a critical factor in the development and persistence of chronic wounds, particularly in the presence of bacterial biofilms.
- Antioxidant interventions show promise for treating chronic wounds by mitigating OS and improving healing outcomes.
- The developed mouse model provides a valuable platform for studying chronic wound pathogenesis and testing novel therapies.
