(verbatim) Bioengineered skin shows great promise in the treatment of chronic wounds that are difficult to heal. Recently, in a prospective randomized trial of 293 patients, we have shown that graftskin, a bilayered skin construct (BSC) composed of neonatal keratinocytes and fibroblasts in type I bovine collagen, accelerates the healing of difficult to heal venous ulcers when compared to compression therapy alone. Yet, we don't know whether the allogeneic cells in BSC persist in the wound and how BSC accelerates healing. Much of the therapeutic effect of BSC is associated with stimulation of the wound's edges toward the center, but it is unclear whether this mechanism involves incorporation of donor cells into the wound margin. Recently, we have found that dermal fibroblasts cultured from the edge of venous ulcers are unresponsive to the action of transforming growth factor-beta-1 (TGF-beta-1), a critical peptide in the formation and deposition of collagen synthesis, and show decreased expression of TGF-beta receptors. The hypothesis of this proposal is that BSC is capable of stimulating the edges of venous ulcers to migrate and can restore the phenotypic make-up of resident wound cells at the margins of the wound. We propose the following specific aims: 1) Determine whether the edges of the wound are stimulated to migrate and whether, by immunohistochemisry and flow cytometry, cells at the edge become activated when BSC is placed only in the center of venous ulcers; 2) Determine by PCR and by fluorescence in situ hybridization whether BSC cells persist in venous ulcers at various times after its application; 3) Measure the response of BSC to injury by the use of flow cytometry, proliferative markers (i.e., ki67) and cytokine expression. 4) Determine whether BSC is capable of restoring responsiveness to TGF-beta-1 and expression of TGF-beta receptors in fibroblasts cultured from non-healing venous ulcers. The proposed studies will advance our understanding of how a bioengineered skin product works in a human chronic wound and provide insight into further bioengineering needs.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR046557-03
Application #
6534492
Study Section
Special Emphasis Panel (ZRG1-SSS-M (01))
Program Officer
Moshell, Alan N
Project Start
2000-09-01
Project End
2004-08-31
Budget Start
2002-09-01
Budget End
2003-08-31
Support Year
3
Fiscal Year
2002
Total Cost
$365,214
Indirect Cost
Name
Roger Williams Hospital
Department
Type
DUNS #
City
Providence
State
RI
Country
United States
Zip Code
02908
Falanga, Vincent; Butmarc, Janet; Cha, Jisun et al. (2007) Migration of the epidermal over the dermal component (epiboly) in a bilayered bioengineered skin construct. Tissue Eng 13:21-8
Cha, Jisun; Falanga, Vincent (2007) Stem cells in cutaneous wound healing. Clin Dermatol 25:73-8
Panuncialman, Jaymie; Falanga, Vincent (2006) Basic approach to inflammatory ulcers. Dermatol Ther 19:365-76
Falanga, Vincent; Saap, Liliana J; Ozonoff, Alexander (2006) Wound bed score and its correlation with healing of chronic wounds. Dermatol Ther 19:383-90
Donohue, Kevin G; Carson, Polly; Iriondo, Manuel et al. (2005) Safety and efficacy of a bilayered skin construct in full-thickness surgical wounds. J Dermatol 32:626-31
Falanga, Vincent (2005) Wound healing and its impairment in the diabetic foot. Lancet 366:1736-43
Falanga, Vincent (2004) The chronic wound: impaired healing and solutions in the context of wound bed preparation. Blood Cells Mol Dis 32:88-94
Butmarc, Janet; Yufit, Tatyana; Carson, Polly et al. (2004) Human beta-defensin-2 expression is increased in chronic wounds. Wound Repair Regen 12:439-43
Falanga, Vincent; Schrayer, David; Cha, Jisun et al. (2004) Full-thickness wounding of the mouse tail as a model for delayed wound healing: accelerated wound closure in Smad3 knock-out mice. Wound Repair Regen 12:320-6
Nahm, Walter K; Philpot, Benjamin D; Adams, Michelle M et al. (2004) Significance of N-methyl-D-aspartate (NMDA) receptor-mediated signaling in human keratinocytes. J Cell Physiol 200:309-17

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