Advertisement

Molecular Medicine

, Volume 13, Issue 1–2, pp 30–39 | Cite as

Molecular Markers in Patients with Chronic Wounds to Guide Surgical Debridement

  • Harold Brem
  • Olivera Stojadinovic
  • Robert F. Diegelmann
  • Hyacinth Entero
  • Brian Lee
  • Irena Pastar
  • Michael Golinko
  • Harvey Rosenberg
  • Marjana Tomic-Canic
Research Article

Abstract

Chronic wounds, such as venous ulcers, are characterized by physiological impairments manifested by delays in healing, resulting in severe morbidity. Surgical debridement is routinely performed on chronic wounds because it stimulates healing. However, procedures are repeated many times on the same patient because, in contrast to tumor excision, there are no objective biological/molecular markers to guide the extent of debridement. To develop bioassays that can potentially guide surgical debridement, we assessed the pathogenesis of the patients’ wound tissue before and after wound debridement. We obtained biopsies from three patients at two locations, the nonhealing edge (prior to debridement) and the adjacent, nonulcerated skin of the venous ulcers (post debridement), and evaluated their histology, biological response to wounding (migration) and gene expression profile. We found that biopsies from the nonhealing edges exhibit distinct pathogenic morphology (hyperproliferative/hyperkeratotic epidermis; dermal fibrosis; increased procollagen synthesis). Fibroblasts deriving from this location exhibit impaired migration in comparison to the cells from adjacent nonulcerated biopsies, which exhibit normalization of morphology and normal migration capacity. The nonhealing edges have a specific, identifiable, and reproducible gene expression profile. The adjacent nonulcerated biopsies have their own distinctive reproducible gene expression profile, signifying that particular wound areas can be identified by gene expression profiling. We conclude that chronic ulcers contain distinct subpopulations of cells with different capacity to heal and that gene expression profiling can be utilized to identify them. In the future, molecular markers will be developed to identify the nonimpaired tissue, thereby making surgical debridement more accurate and more efficacious.

Notes

Acknowledgments

Special thanks to Dr H. Paul Ehrlich, Dr Stephen Dotty, and members of HSS histology core for their assistance in histology analyses. Our research is supported by the National Institutes of Health grants DK59424 (HB), LM008443 (HB), AR45974 (MT-C), NR08029 (MT-C), and the VCU A D Williams Foundation (RFD). We also acknowledge the use of The Musculoskeletal Repair and Regeneration Core Center of HSS (AR046121).

References

  1. 1.
    Brem H et al. (2003) Healing of elderly patients with diabetic foot ulcers, venous stasis ulcers, and pressure ulcers. Surg. Technol. Int. 11:161–167.PubMedGoogle Scholar
  2. 2.
    Bergan JJ et al. (2006) Chronic venous disease. N. Engl. J. Med. 355:488–498.CrossRefGoogle Scholar
  3. 3.
    Services UDoHaH. (2004) Guidance to surveyors for long term care facilities. Guidance to Surveyors for Long Term Care Facilities on World Wide Web. URL: https://doi.org/www.cms.hhs.gov//.
  4. 4.
    Steed DL, Donohoe D, Webster MW, Lindsley L, Diabetic Ulcer Study Group. (1996) Effect of extensive debridement and treatment on the healing of diabetic foot ulcers. J. Am. Coll. Surg. 77:575–586.Google Scholar
  5. 5.
    Sibbald RG et al. (2000) Preparing the wound bed—debridement, bacterial balance, and moisture balance. Ostomy Wound Manage. 46:14–22, 24–18, 30–15; quiz 36–17.PubMedGoogle Scholar
  6. 6.
    Brem H, Balledux J, Sukkarieh T, Carson P, Falanga V. (2001) Healing of venous ulcers of long duration with a bilayered living skin substitute: results from a general surgery and dermatology department. Dermatol. Surg. 27:915–919.PubMedGoogle Scholar
  7. 7.
    Falanga V et al. (1998) Rapid healing of venous ulcers and lack of clinical rejection with an allogeneic cultured human skin equivalent. Arch. Dermatol. 134:293–300.CrossRefGoogle Scholar
  8. 8.
    Falanga V, Sabolinski M. (1999) A bilayered living skin construct (APLIGRAF) accelerates complete closure of hard-to-heal venous ulcers. Wound Repair Regen 7:201–207.CrossRefGoogle Scholar
  9. 9.
    Stojadinovic O et al. (2005) Molecular pathogenesis of chronic wounds: the role of beta-catenin and c-myc in the inhibition of epithelialization and wound healing. Am. J. Pathol. 167:59–69.CrossRefGoogle Scholar
  10. 10.
    Davies CE, Turton G, Woolfrey G, Elley R, Taylor M. (2005) Exploring debridement options for chronic venous leg ulcers. Br. J. Nurs. 14:393–397.CrossRefGoogle Scholar
  11. 11.
    Brem H, Lyder C. (2004) Protocol for the successful treatment of pressure ulcers. Am. J. Surg. 188:9–17.CrossRefGoogle Scholar
  12. 12.
    Golub TR et al. (1999) Molecular classification of cancer: class discovery and class prediction by gene expression monitoring. Science 286:531–537.CrossRefGoogle Scholar
  13. 13.
    Risinger JI et al. (2003) Microarray analysis reveals distinct gene expression profiles among different histologic types of endometrial cancer. Cancer Res. 63:6–11.PubMedGoogle Scholar
  14. 14.
    Van de Vijver MJ et al. (2002) A gene-expression signature as a predictor of survival in breast cancer. N. Engl. J. Med. 347:1999–2009.CrossRefGoogle Scholar
  15. 15.
    Haider AS et al. (2006) Genomic analysis defines a cancer-specific gene expression signature for human squamous cell carcinoma and distinguishes malignant hyperproliferation from benign hyperplasia. J. Invest. Dermatol. 126:869–881.CrossRefGoogle Scholar
  16. 16.
    Grose R. (2004) Common ground in the transcriptional profiles of wounds and tumors. Genome Biol. 5:228.CrossRefGoogle Scholar
  17. 17.
    Wang Y. (2005) Gene expression-driven diagnostics and pharmacogenomics an cancer. Curr.Opin. Mol. Ther. 7:246–250.PubMedGoogle Scholar
  18. 18.
    Cao Z, Wu HK, Bruce A, Wollenberg K, Panjwani N. (2002) Detection of differentially expressed genes in healing mouse corneas, using cDNA microarrays. Invest. Ophthalmol. Vis. Sci. 43:2897–2904.PubMedGoogle Scholar
  19. 19.
    Nakazawa T et al. (2004) Gene expression of periostin in the early stage of fracture healing detected by cDNA microarray analysis. J. Orthop. Res. 22:520–525.CrossRefGoogle Scholar
  20. 20.
    Cole J, Tsou R, Wallace K, Gibran N, Isik F. (2001) Early gene expression profile of human skin to injury using high-density cDNA microarrays. Wound Repair Regen. 9:360–370.CrossRefGoogle Scholar
  21. 21.
    McDonald JA et al. (1986) A monoclonal antibody to the carboxyterminal domain of procollagen type I visualizes collagen-synthesizing fibroblasts. Detection of an altered fibroblast phenotype in lungs of patients with pulmonary fibrosis. J. Clin. Invest. 78:1237–1244.CrossRefGoogle Scholar
  22. 22.
    Stojadinovic O et al. (2007) Novel genomic effects of glucocorticoids in epidermal keratinocytes: Inhibition of apoptosis, IFNgamma pathway and wound healing along with promotion of terminal differentiation. J Biol Chem. 282:4021–4034.CrossRefGoogle Scholar
  23. 23.
    Radoja N, Komine M, Jho SH, Blumenberg M, Tomic-Canic M. (2000) Novel mechanism of steroid action in skin through glucocorticoid receptor monomers. Mol. Cell Biol. 20:4328–4339.CrossRefGoogle Scholar
  24. 24.
    Lee B, Vouthounis C, Stojadinovic O, Brem H, Im M, Tomic-Canic M. (2005) From an enhanceosome to a repressosome: molecular antagonism between glucocorticoids and EGF leads to inhibition of wound healing. J. Mol. Biol. 345:1083–1097.CrossRefGoogle Scholar
  25. 25.
    De Heller-Milev M, Huber M, Panizzon R, Hohl D. (2000) Expression of small proline rich proteins in neoplastic and inflammatory skin diseases. Br. J. Dermatol. 143:733–740.CrossRefGoogle Scholar
  26. 26.
    Freedberg IM, Tomic-Canic M, Komine M, Blumenberg M. (2001) Keratins and the keratinocyte activation cycle. J. Invest. Dermatol. 116:633–640.CrossRefGoogle Scholar
  27. 27.
    Lobmann R et al. (2002) Expression of matrix-metalloproteinases and their inhibitors in the wounds of diabetic and non-diabetic patients. Diabetologia 45:1011–1016.CrossRefGoogle Scholar
  28. 28.
    Egawa K, Honda Y, Ono T, Kuroki M. (1998) Immunohistochemical demonstration of carcinoembryonic antigen and related antigens in various cutaneous keratinous neoplasms and verruca vulgaris. Br. J. Dermatol. 139:178–185.CrossRefGoogle Scholar
  29. 29.
    Wang H et al. (2003) Expression of insulin-like growth factor-binding protein 2 in melanocytic lesions. J. Cutan. Pathol. 30:599–605.CrossRefGoogle Scholar
  30. 30.
    Kansal RG, Aziz RK, Kotb M. (2005) Modulation of expression of superantigens by human transferrin and lactoferrin: a novel mechanism in host-Streptococcus interactions. J. Infect. Dis. 191:2121–2129.CrossRefGoogle Scholar
  31. 31.
    Martinez-Esparza M, Jimenez-Cervantes C, Solano F, Lozano JA, Garcia-Borron JC. (1998) Mechanisms of melanogenesis inhibition by tumor necrosis factor-alpha in B16/F10 mouse melanoma cells. Eur. J. Biochem. 255:139–146.CrossRefGoogle Scholar
  32. 32.
    Hu P et al. (2002) Role of membrane proteins in permeability barrier function: uroplakin ablation elevates urothelial permeability. Am. J. Physiol. Renal Physiol. 283:F1200–1207.CrossRefGoogle Scholar
  33. 33.
    Bowen GM, White GLJ, Gerwels JW. (2005) Mohs microscopic surgery. Am. Fam. Physician 72:845–848.PubMedGoogle Scholar
  34. 34.
    Williams D et al. (2005) Effect of sharp debridement using curette on recalcitrant nonhealing venous leg ulcers: a concurrently controlled, prospective cohort study. Wound Repair Regen. 13:131–137.CrossRefGoogle Scholar
  35. 35.
    Falanga V, Eaglstein WH. (1993) The “trap” hypothesis of venous ulceration. Lancet 341:1006–1008.CrossRefGoogle Scholar
  36. 36.
    Van de Scheur M, Falanga V. (1997) Pericapillary fibrin cuffs in venous disease. A reappraisal. Dermatol. Surg. 23:955–959.CrossRefGoogle Scholar
  37. 37.
    Tomic-Canic M, Komine M, Freedberg IM, Blumenberg M. (1998) Epidermal signal transduction and transcription factor activation in activated keratinocytes. J. Dermatol. Sci. 17:167–181.CrossRefGoogle Scholar
  38. 38.
    Roh JY, Stanley JR. (1995) Plakoglobin binding by human Dsg3 (pemphigus vulgaris antigen) in keratinocytes requires the cadherin-like intracytoplasmic segment. J. Invest. Dermatol. 104:720–724.CrossRefGoogle Scholar
  39. 39.
    Dellambra E et al. (1995) Stratifin, a keratinocyte specific 14-3-3 protein, harbors a pleckstrin homology (PH) domain and enhances protein kinase C activity. J. Cell Sci. 108 (Pt 11):3569–3579.PubMedGoogle Scholar
  40. 40.
    Cyranoski D. (2005) Japan jumps toward personalized medicine. Nature 437:796.CrossRefGoogle Scholar
  41. 41.
    Varmus H. (2006) The new era in cancer research. Science 312:1162–1165.CrossRefGoogle Scholar

Copyright information

© Feinstein Institute for Medical Research 2007

Authors and Affiliations

  • Harold Brem
    • 1
  • Olivera Stojadinovic
    • 2
  • Robert F. Diegelmann
    • 3
  • Hyacinth Entero
    • 1
    • 4
  • Brian Lee
    • 5
  • Irena Pastar
    • 2
  • Michael Golinko
    • 1
  • Harvey Rosenberg
    • 1
  • Marjana Tomic-Canic
    • 2
  1. 1.Department of Surgery, Wound Healing and Vascular Biology LaboratoryColumbia University College of Physicians and SurgeonsNew YorkUSA
  2. 2.Tissue Repair Lab, Tissue Engineering, Regeneration and Repair ProgramHospital for Special Surgery of the Weill Cornell College of MedicineNew YorkUSA
  3. 3.Department of BiochemistryVirginia Commonwealth UniversityRichmondUSA
  4. 4.Ross University School of MedicineRoseauCommonwealth of Dominica
  5. 5.GenentechSan FranciscoUSA

Personalised recommendations