Wounding of bioengineered skin: Cellular and molecular aspects after injury

Vincent Falanga, Cary Isaacs, Dana Paquette, Gregory Downing, Nicola Kouttab, Janet Butmarc, Evangelos V Badiavas, Jan Hardin-Young

Research output: Contribution to journalArticle

80 Citations (Scopus)

Abstract

Skin substitutes are increasingly being used in the treatment of difficult to heal wounds but their mechanisms of action are largely unknown. In this study, using histology, immunostaining, flow cytometry, enzyme-linked immunosorbent assay, and reverse transcription polymerase chain reaction, we determined the response to injury of a human bilayered skin substitute. Meshing or scalpel fenestration of the construct was found to stimulate keratinocyte migration and to decrease proliferation. By 24 h, flow cytometry of the keratinocyte component showed that meshing was associated with a 33% decrease in the number of cells in S phase (p < 0.01). An approximately 2-fold decrease in staining for Ki67, a proliferation marker, was observed with meshing of human bilayered skin substitute. The process of reepithelialization was apparent by 12 h, however, the wounded human bilayered skin substitute was healed by day 3, and a stratum corneum and fully stratified epithelium were re-established by day 4. Reverse transcription polymerase chain reaction analysis and enzyme-linked immunosorbent assays showed that the expression of acute proinflammatory cytokines (interleukins 1α, 6, and 8, tumor necrosis factor α) peaked by 12-24 h postinjury. The levels of mRNA of certain growth factors (transforming growth factor β1, vascular endothelial growth factor, insulin-like growth factor 2) but not others (platelet-derived growth factors A and B, keratinocyte growth factor, fibroblast growth factors 1 and 7, transforming growth factor β3) increased by 12 h and peaked by 1-3 d after injury, returning to normal by day 6. Immunostaining for tumor necrosis factor α and transforming growth factor β1 paralleled these findings by reverse transcription polymerase chain reaction. We conclude that human bilayered skin substitute, as a prototypic bilayered skin substitute, is a truly dynamic living tissue, capable of responding to physical injury in a staged and specific pattern of cell migration, reepithelialization, and cytokine expression.

Original languageEnglish
Pages (from-to)653-660
Number of pages8
JournalJournal of Investigative Dermatology
Volume119
Issue number3
DOIs
StatePublished - Oct 9 2002
Externally publishedYes

Fingerprint

Artificial Skin
Skin
Polymerase chain reaction
Wounds and Injuries
Transforming Growth Factors
Transcription
Fibroblast Growth Factor 7
Reverse Transcription
Immunosorbents
Flow cytometry
Keratinocytes
Polymerase Chain Reaction
Assays
Flow Cytometry
Tumor Necrosis Factor-alpha
Enzyme-Linked Immunosorbent Assay
Proto-Oncogene Proteins c-sis
Cytokines
Fibroblast Growth Factor 1
Histology

Keywords

  • Cytokines
  • Growth factors
  • Keratocytes
  • Skin substitute
  • Tissue repair
  • Wound healing

ASJC Scopus subject areas

  • Dermatology

Cite this

Falanga, V., Isaacs, C., Paquette, D., Downing, G., Kouttab, N., Butmarc, J., ... Hardin-Young, J. (2002). Wounding of bioengineered skin: Cellular and molecular aspects after injury. Journal of Investigative Dermatology, 119(3), 653-660. https://doi.org/10.1046/j.1523-1747.2002.01865.x

Wounding of bioengineered skin : Cellular and molecular aspects after injury. / Falanga, Vincent; Isaacs, Cary; Paquette, Dana; Downing, Gregory; Kouttab, Nicola; Butmarc, Janet; Badiavas, Evangelos V; Hardin-Young, Jan.

In: Journal of Investigative Dermatology, Vol. 119, No. 3, 09.10.2002, p. 653-660.

Research output: Contribution to journalArticle

Falanga, V, Isaacs, C, Paquette, D, Downing, G, Kouttab, N, Butmarc, J, Badiavas, EV & Hardin-Young, J 2002, 'Wounding of bioengineered skin: Cellular and molecular aspects after injury', Journal of Investigative Dermatology, vol. 119, no. 3, pp. 653-660. https://doi.org/10.1046/j.1523-1747.2002.01865.x
Falanga, Vincent ; Isaacs, Cary ; Paquette, Dana ; Downing, Gregory ; Kouttab, Nicola ; Butmarc, Janet ; Badiavas, Evangelos V ; Hardin-Young, Jan. / Wounding of bioengineered skin : Cellular and molecular aspects after injury. In: Journal of Investigative Dermatology. 2002 ; Vol. 119, No. 3. pp. 653-660.
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