JoVE Logo
Faculty Resource Center

Sign In

Summary

Abstract

Introduction

Protocol

Representative Results

Discussion

Acknowledgements

Materials

References

Medicine

Murine Model of Wound Healing

Published: May 28th, 2013

DOI:

10.3791/50265

1The Heart Research Institute, 2Sydney Medical School, University of Sydney , 3Cardiology Department, Royal Prince Alfred Hospital

A murine model of cutaneous wound healing that can be used to assess therapeutic compounds in physiological and pathophysiological settings.

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. Impaired wound healing, which occurs in diabetic patients for example, can lead to severe unfavorable outcomes such as amputation. There is, therefore, an increasing impetus to develop novel agents that promote wound repair. The testing of these has been limited to large animal models such as swine, which are often impractical. Mice represent the ideal preclinical model, as they are economical and amenable to genetic manipulation, which allows for mechanistic investigation. However, wound healing in a mouse is fundamentally different to that of humans as it primarily occurs via contraction. Our murine model overcomes this by incorporating a splint around the wound. By splinting the wound, the repair process is then dependent on epithelialization, cellular proliferation and angiogenesis, which closely mirror the biological processes of human wound healing. Whilst requiring consistency and care, this murine model does not involve complicated surgical techniques and allows for the robust testing of promising agents that may, for example, promote angiogenesis or inhibit inflammation. Furthermore, each mouse acts as its own control as two wounds are prepared, enabling the application of both the test compound and the vehicle control on the same animal. In conclusion, we demonstrate a practical, easy-to-learn, and robust model of wound healing, which is comparable to that of humans.

Impaired wound healing is responsible for significant morbidity and mortality worldwide; this is particularly true for sufferers of diabetes mellitus1,2. In humans, wound healing is a continuum of processes, in which there is significant overlapping3. Immediately following wounding, inflammatory processes are initiated. Inflammatory cells release factors that encourage the processes of cell proliferation, migration and angiogenesis. After re-epithelialization and new tissue formation there is a phase of remodeling that entails both apoptosis and re-organization of matrix proteins such as collagen.

The complexity of wou....

Log in or to access full content. Learn more about your institution’s access to JoVE content here

1. Preparation of Splints and Occlusive Dressings

  1. Outline 10 mm circles on 0.5 mm thick silicone sheeting and use scissors or a biopsy punch to create silicone disks.
  2. Centre a 5 mm biopsy punch in the middle of the 10 mm circle and press firmly to create a hole to form a "donut"-like disc that will be used as a splint.
  3. Outline 10 mm circles on a transparent occlusive dressing such as Opsite and use scissors to create circular dressings.

2. Experimental Anim.......

Log in or to access full content. Learn more about your institution’s access to JoVE content here

A wound closure curve is determined by calculating the average diameter of the wound and expressing the results as a percentage, i.e. 100 - (Day 0 diameter/Day X diameter). In this experiment a therapeutic compound (or vehicle control) was applied daily to the wound. The therapeutic compound greatly accelerated wound closure (Figure 3). It is important to note that the splints must be maintained for the duration of the experiment, as removal of splints will lead to rapid wound contraction (

Log in or to access full content. Learn more about your institution’s access to JoVE content here

This is an experimental murine model of cutaneous wound healing. A significant feature of this model is the use of silicone splints to prevent wound contraction so that re-epithelialization and new tissue formation may occur, making it similar to the process that occurs in humans. This model is versatile and can be used to assess wound healing in both physiological and pathophysiological (e.g. diabetes mellitus) settings. The model may also be used to assess potential wound healing or angiogenesis therapeutics i.......

Log in or to access full content. Learn more about your institution’s access to JoVE content here

The authors would like to acknowledge funding support from the National Health and Medical Research Council (NHMRC) of Australia (Project Grant ID: 632512). Louise Dunn was supported by an NHMRC Early Career Fellowship and Christina Bursill by a National Heart Foundation Career Development Fellowship.

....

Log in or to access full content. Learn more about your institution’s access to JoVE content here

Name Company Catalog Number Comments
Name of Reagent/Material Company Catalog Number Comments
Press-to-seal silicone sheeting 0.5 mm thick Invitrogen P18178 Cut into "donuts" with external diameter of 1cm external, 0.5 cm internal diameter
Biopsy punch 5 mm Steifel BC-B1-0500 To outline wound area to be excised
Vannas scissors 8.5 cm curved World Precision Instruments 501232 For wound incision and excision
Dumonte #7b forceps, 11 cm World Precision Instruments 501302 To grip skin when creating incision and excising skin
Graefe forceps, serrated 10cm World Precision Instruments 14142 To help attach silicone splint to skin
Needle holder, smooth jaws, curved, 12.5 cm World Precision Instruments 14132
Malis forceps, smooth, straight, 12 cm Codman and Shurtleff, Inc (J&J) 80-1500 To suture the silicon rings to the skin
Ruler, 0.5 mm gradation n/a
Calipers 0.25 mm gradation Duckworth and Kent 9-653 To measure wound area
Opsite FlexiFix transparent adhesive film. 10 cm x 1 m Smith & Nephew 66030570
Rimadyl (Carprofen) Pfizer 462986

  1. Sen, C. K., et al. Human skin wounds: a major and snowballing threat to public health and the economy. Wound Repair. 17, 763-771 (2009).
  2. Sen, C. K. Wound healing essentials: let there be oxygen. Wound Repair Regen. 17, 1-18 (2009).
  3. Gurtner, G. C., Werner, S., Barrandon, Y. Wound repair and regeneration. Nature. 453, 314-321 (2008).
  4. Lindblad, W. J. Considerations for selecting the correct animal model for dermal wound-healing studies. J. Biomater. Sci. Polym. Ed. 19, 1087-1096 (2008).
  5. Grose, R., Werner, S. Wound-healing studies in transgenic and knockout mice. Mol. Biotechnol. 28, 147-166 (2004).
  6. Reid, R. R., Said, H. K., Mogford, J. E., Mustoe, T. A. The future of wound healing: pursuing surgical models in transgenic and knockout mice. J. Am. Coll. Surg. 199, 578-585 (2004).
  7. Fang, R. C., Mustoe, T. A. Animal models of wound healing: utility in transgenic mice. J. Biomater. Sci. Polym. Ed. 19, 989-1005 (2008).
  8. Wong, V. W., Sorkin, M., Glotzbach, J. P., Longaker, M. T., Gurtner, G. C. Surgical approaches to create murine models of human wound healing. J. Biomed. Biotechnol. 2011, 969618 (2011).
  9. Galiano, R. D., Michaels, J. t., Dobryansky, M., Levine, J. P., Gurtner, G. C. Quantitative and reproducible murine model of excisional wound healing. Wound Repair. 12, 485-492 (2004).
  10. Galiano, R. D., et al. Topical vascular endothelial growth factor accelerates diabetic wound healing through increased angiogenesis and by mobilizing and recruiting bone marrow-derived cells. The American Journal of Pathology. 164, 1935-1947 (2004).
  11. Thangarajah, H., et al. The molecular basis for impaired hypoxia-induced VEGF expression in diabetic tissues. Proceedings of the National Academy of Sciences of the United States of America. 106, 13505-13510 (2009).
  12. Raza, A., Bayles, C., Biebel, D. Investigation of Bacterial Growth and Moisture Handling Properties of Transparent Dressings: 3M Tegaderm Transparent Dressing, 3M Tegaderm HP Transparent Dressing, and Opsite IV3000 Transparent Dressing. Smith and Nephew Report. , (1998).
  13. Chung, T. Y., Peplow, P. V., Baxter, G. D. Testing photobiomodulatory effects of laser irradiation on wound healing: development of an improved model for dressing wounds in mice. Photomed. Laser Surg. 28, 589-596 (2010).

This article has been published

Video Coming Soon

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2024 MyJoVE Corporation. All rights reserved