Retrovirally mediated gene transfer in a skin equivalent model of chronic wounds

Evangelos V Badiavas, Parmender P. Mehta, Vincent Falanga

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Given that treatment for chronic wounds is unsatisfactory, it is likely that gene therapy may be tested as a therapeutic modality in this difficult clinical problem. Actively proliferating cells in wounds are also a good target for retroviral transduction, an increasingly useful method for gene therapy. However, it is unclear how gene therapy may best be used in chronic wounds, and experimental models are urgently needed to study and manipulate gene transfer in the context of chronic wounds. In this report, partial- and full-thickness wounds were made in vitro in a human living skin equivalent (LSE) consisting of fully differentiated keratinocytes layered over a collagen matrix seeded with fibroblasts. To mimic a chronic wound situation, we used tissue culture conditions which, as in a chronic wound, allowed fibroblast but not keratinocyte proliferation or migration. The wounded LSE was then placed over a transduced cell line (PA317) which produced a replication defective retrovirus containing as a histological marker the bacterial beta galactosidase gene. Using this close and direct exposure to the virus-producing cell line, distinct staining for β-galactosidase was observed in partial-thickness wounds, and was limited to fibroblasts away from the upper site of injury and immediately overlying the retrovirus-producing cell monolayer. Expression of β-galactosidase was uniformly present at the wound edges and along the base of the entire partial thickness wound. These studies demonstrate that, in in vivo conditions mimicking a chronic wound, an intimate apposition of the injured LSE with the virus-producing cell line is needed for gene transfer. Using this in vitro model system, gene transfer protocols may be optimized prior to beginning in vivo studies in chronic wounds.

Original languageEnglish
Pages (from-to)56-62
Number of pages7
JournalJournal of Dermatological Science
Volume13
Issue number1
DOIs
StatePublished - Oct 1 1996

Fingerprint

Gene transfer
Gene therapy
Fibroblasts
Galactosidases
Skin
Cells
Viruses
Wounds and Injuries
Genes
Tissue culture
beta-Galactosidase
Monolayers
Collagen
Genetic Therapy
Retroviridae
Keratinocytes
Cell Line

Keywords

  • Gene
  • Retrovirus
  • Skin
  • Therapy
  • Wounds

ASJC Scopus subject areas

  • Dermatology

Cite this

Retrovirally mediated gene transfer in a skin equivalent model of chronic wounds. / Badiavas, Evangelos V; Mehta, Parmender P.; Falanga, Vincent.

In: Journal of Dermatological Science, Vol. 13, No. 1, 01.10.1996, p. 56-62.

Research output: Contribution to journalArticle

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