Skip to main content
SpringerLink
Log in
Book cover

Wound Regeneration and Repair pp 203–231Cite as

Novel Methods for the Investigation of Human Hypertrophic Scarring and Other Dermal Fibrosis

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1037))

Abstract

Hypertrophic scar (HTS) represents the dermal equivalent of fibroproliferative disorders that occur after injury involving the deep dermis while superficial wounds to the skin heal with minimal or no scarring. HTS is characterized by progressive deposition of collagen that occurs with high frequency in adult dermal wounds following traumatic or thermal injury. Increased levels of transforming growth factor-β1 (TGF-β1), decreased expression of small leucine-rich proteoglycans (SLRPs), and/or fibroblast subtypes may influence the development of HTS. The development of HTS is strongly influenced by the cellular and molecular properties of fibroblast subtypes, where cytokines such as fibrotic TGF-β1 and CTGF as well as the expression of SLRPs, particularly decorin and fibromodulin, regulate collagen fibrillogenesis and the activity of TGF-β1. Reduced anti-fibrotic molecules in the ECM of the deep dermis and the distinctive behavior of the fibroblasts in this region of the dermis which display increased sensitivity to TGF-β1’s biological activity contribute to the development of HTS following injury to the deep dermis. By comparing the cellular and molecular differences involved in deep and superficial wound healing in an experimental wound scratch model in humans that has both superficial and deep injuries within the same excisional model, our aim is to increase our understanding of how tissue repair following injury to the deep dermis can be changed to promote healing with a similar pattern to healing that occurs following superficial injury that results in no or minimal scarring. Studying the characteristics of superficial dermal injuries that heal with minimal scarring will help us identify therapeutic approaches for tissue engineering and wound healing. In addition, our ability to develop novel therapies for HTS is hampered by limitations in the available animal models used to study this disorder in vivo. We also describe a nude mouse model of transplanted human skin that develops a hypertrophic proliferative scar consistent morphologically and histologically with human HTS, which can be used to test novel treatment options for these dermal fibrotic conditions.

This is a preview of subscription content, log in via an institution.

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   159.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Wang J, Dodd C, Shankowsky HA et al (2008) Deep dermal fibroblasts contribute to hypertrophic scarring. Lab Invest 88:1278

    Article  CAS  PubMed  Google Scholar 

  2. Gallant CL, Olson ME, Hart DA (2004) Molecular, histologic and gross phenotype of skin wound healing in red Duroc pigs reveals an abnormal healing phenotype of hypercontracted, hyperpigmented scarring. Wound Repair Regen 12:305

    Article  PubMed  Google Scholar 

  3. Sappino AP, Masouye I, Saurat JH et al (1990) Smooth muscle differentiation in scleroderma fibroblastic cells. Am J Pathol 137:585

    CAS  PubMed Central  PubMed  Google Scholar 

  4. Wrana JL, Attisano L, Cárcamo J et al (1992) TGF-beta signals through a heteromeric protein kinase receptor complex. Cell 71:1003

    Article  CAS  PubMed  Google Scholar 

  5. Souchelnytskyi S, Rönnstrand L, Heldin CH et al (2001) Phosphorylation of Smad signalling proteins by receptor serine/threonine kinases. Methods Mol Biol 124:107

    CAS  PubMed  Google Scholar 

  6. Hildebrand A, Romaris M, Rasmussen LM et al (1994) Interaction of the small interstitial proteoglycans biglycan, decorin and fibromodulin with transforming growth factor beta. Biochem J 302:527

    CAS  PubMed Central  PubMed  Google Scholar 

  7. Soo C, Hu FY, Zhang X et al (2000) Differential expression of fibromodulin, a transforming growth factor-beta modulator, in fetal skin development and scarless repair. Am J Pathol 157:423

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  8. Kolb M, Margetts PJ, Sime PJ et al (2001) Proteoglycans decorin and biglycan differentially modulate TGFβ-mediated fibrotic responses in the lung. Am J Physiol Lung Cell Mol Physiol 280:1327

    Google Scholar 

  9. Sorrell JM, Baber MA, Caplan AI (2004) Site-matched papillary and reticular human dermal fibroblasts differ in their release of specific growth factors/cytokines and in their interaction with keratinocytes. J Cell Physiol 200:134

    Article  CAS  PubMed  Google Scholar 

  10. Hagood JS, Lasky JA, Nesbitt JE et al (2001) Differential expression, surface binding, and response to connective tissue growth factor in lung fibroblast subpopulations. Chest 120:64

    Article  Google Scholar 

  11. Zhou Y, Hagood JS, Murphy-Ullrich JE (2004) Thy-1 expression regulates the ability of rat lung fibroblasts to activate transforming growth factor-beta in response to fibrogenic stimuli. Am J Pathol 165:659

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  12. Hagood JS, Prabhakaran P, Kumbla P et al (2005) Loss of fibroblast Thy-1 expression correlates with lung fibrogenesis. Am J Pathol 167:365

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  13. Dunkin CS, Pleat JM, Gillespie PH et al (2007) Scarring occurs at a critical depth of skin injury: precise measurement in a graduated dermal scratch in human volunteers. Plast Reconstr Surg 119:1722

    Article  CAS  PubMed  Google Scholar 

  14. Honardoust D, Varkey M, Hori K et al (2011) Small leucine-rich proteoglycans, decorin and fibromodulin, are reduced in postburn hypertrophic scar. Wound Repair Regen 19:368–378

    Article  PubMed  Google Scholar 

  15. Ali-Bahar M, Bauer B, Tredget EE, Ghahary A (2004) Dermal fibroblasts from different layers of human skin are heterogeneous in expression of collagenase and types I and III procollagen mRNA. Wound Repair Regen 12(2):175–182

    Article  PubMed  Google Scholar 

  16. Kitano Y, Okada N (1983) Separation of the epidermal sheet by dispase. Br J Dermatol 108(5):555–560

    Article  CAS  PubMed  Google Scholar 

  17. Schönherr E, Beavan LA, Hausser H, Kresse H, Culp LA (1993) Differences in decorin expression by papillary and reticular fibroblasts in vivo and in vitro. Biochem J 290(Pt 3):893–899

    PubMed Central  PubMed  Google Scholar 

  18. Mustoe TA, Cooter RD, Gold MH et al (2002) International clinical recommendations on scar management. Plast Reconstr Surg 110:560–571

    Article  PubMed  Google Scholar 

  19. Scott PG, Dodd CM, Tredget EE et al (1995) Immunohistochemical localization of the proteoglycans decorin, biglycan and versican and transforming growth factor-beta in human post-burn hypertrophic and mature scars. Histopathology 26:423

    Article  CAS  PubMed  Google Scholar 

  20. Oliveira GV, Hawkins HK, Chinkes D et al (2009) Hypertrophic versus non hypertrophic scars compared by immunohistochemistry and laser confocal microscopy: type I and III collagens. Int Wound J 6:445

    Article  PubMed Central  PubMed  Google Scholar 

  21. Mohan RR, Gupta R, Mehan MK et al (2010) Decorin transfection suppresses profibrogenic genes and myofibroblast formation in human corneal fibroblasts. Exp Eye Res 91:238

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  22. Stoff A, Rivera AA, Mathis JM et al (2007) Effect of adenoviral mediated overexpression of fibromodulin on human dermal fibroblasts and scar formation in full-thickness incisional wounds. J Mol Med 85:481

    Article  CAS  PubMed  Google Scholar 

  23. Shah M, Foreman DM, Ferguson MW (1995) Neutralization of TGF-β 1 and TGF-β 2 or exogenous addition of TGF-β 3 to cutaneous rat wounds reduces scarring. J Cell Sci 108:985

    CAS  PubMed  Google Scholar 

  24. Pohlers D, Brenmoehl J, Löffler I et al (2009) TGF-beta and fibrosis in different organs—molecular pathway imprints. Biochim Biophys Acta 1792:746

    Article  CAS  PubMed  Google Scholar 

  25. Bandyopadhyay B, Fan J, Guan S, Li Y et al (2006) A “traffic control” role for TGF-beta3: orchestrating dermal and epidermal cell motility during wound healing. J Cell Biol 172:1093

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  26. Honardoust D, Eslami A, Larjava H, Häkkinen L (2008) Localization of small leucine-rich proteoglycans and transforming growth factor-beta in human oral mucosal wound healing. Wound Repair Regen 16:814

    Article  PubMed  Google Scholar 

  27. Colwell AS, Yun R, Krummel TM et al (2007) Keratinocytes modulate fetal and postnatal fibroblast transforming growth factor-beta and Smad expression in co-culture. Plast Reconstr Surg 119:1440

    Article  CAS  PubMed  Google Scholar 

  28. Goldberg SR, McKinstry RP, Sykes V et al (2007) Rapid closure of midgestational excisional wounds in a fetal mouse model is associated with altered transforming growth factor-beta isoform and receptor expression. J Pediatr Surg 42:966

    Article  PubMed  Google Scholar 

  29. Keeley EC, Mehrad B, Strieter RM (2009) The role of circulating mesenchymal progenitor cells (fibrocytes) in the pathogenesis of fibrotic disorders. Thromb Haemost 101:613

    CAS  PubMed Central  PubMed  Google Scholar 

  30. Yang L, Scott PG, Dodd C et al (2005) Identification of fibrocytes in postburn hypertrophic scar. Wound Repair Regen 13:398

    Article  PubMed  Google Scholar 

  31. Wang J, Jiao H, Stewart TL et al (2007) Improvement in postburn hypertrophic scar after treatment with IFN-alpha2b is associated with decreased fibrocytes. J Interferon Cytokine Res 27:921

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

The wound healing research in the author’s laboratory is funded by the Canadian Institutes of Health Research, the Alberta Heritage Foundation for Medical Research, and Firefighters’ Burn Trust Fund of the University of Alberta.

Author information

Authors and Affiliations

  1. Wound Healing Research Group, Plastic Surgery Research Laboratory, Department of Surgery, University of Alberta, Edmonton, AB, Canada

    Dariush Honardoust, Jie Ding & Edward E. Tredget

  2. Firefighters’ Burn Treatment Unit, Division of Plastic Surgery and Reconstructive Surgery, University of Alberta, Edmonton, AB, Canada

    Peter Kwan, Moein Momtazi & Edward E. Tredget

  3. Division of Critical Care Medicine, University of Alberta, Edmonton, AB, Canada

    Peter Kwan, Moein Momtazi & Edward E. Tredget

  4. Department of Surgery, University of Alberta, Edmonton, AB, Canada

    Peter Kwan, Moein Momtazi & Edward E. Tredget

Authors
  1. Dariush Honardoust
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Peter Kwan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Moein Momtazi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jie Ding
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Edward E. Tredget
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Virginia Tech Carilion Research Institut, Roanoke, Virginia, USA

    Robert G. Gourdie

  2. School of Pharmacy, MCPHS University, Worcester, Massachusetts, USA

    Tereance A. Myers

Rights and permissions

Reprints and permissions

Copyright information

© 2013 Springer Science+Business Media New York

About this protocol

Cite this protocol

Honardoust, D., Kwan, P., Momtazi, M., Ding, J., Tredget, E.E. (2013). Novel Methods for the Investigation of Human Hypertrophic Scarring and Other Dermal Fibrosis. In: Gourdie, R., Myers, T. (eds) Wound Regeneration and Repair. Methods in Molecular Biology, vol 1037. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-505-7_11

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-505-7_11

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-504-0

  • Online ISBN: 978-1-62703-505-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation