Role of tumor necrosis factor-alpha in wound repair in human vocal fold fibroblasts.
OBJECTIVE
Tumor necrosis factor-alpha (TNF-alpha) is an inflammatory cytokine and apoptotic molecule that appears to be a mediator in inflammation and fibrosis. The objective of this investigation was to examine the effects of TNF-alpha on 3D Carbylan-GSX in vitro cultured human vocal fold fibroblasts (hVFFs), to provide insight into the mechanism responsible for the improved vocal fold wound healing that has been previous reported with Carbylan-GSX treatment.


METHODS
In vitro cell culture.


METHODS
hVFF were cultured in 3D Carbylan-GSX and on polystyrene with different dosages of TNF-alpha (0, 0.1, 1, 10, and 100 ng/mL) with and without 10% fetal bovine serum (FBS). hVFF response to TNF-alpha was characterized by morphology, proliferation rates, and gene transcript levels for matrix metalloproteinase 1 (MMP1), matrix metalloproteinase 2 (MMP2), tissue inhibitor of metalloproteinase 3 (TIMP3), collagen I, collagen III, fibronectin, and TNF-alpha receptor.


RESULTS
In 3D Carbylan-GSX, TNF-alpha inhibited hVFF proliferation in a dose-dependent manner. TNF-alpha (0.1-100 ng/mL) was shown to significantly downregulate TIMP3 and extracellular matrix-related mRNA transcript levels for collagen III and fibronectin and to upregulate MMP1 and MMP2 expression, resulting in increased MMP/TIMP3 ratios. TNF-alpha receptor expression was significantly upregulated in Carbylan-GSX compared to control polystyrene. Responses were more marked in 10% FBS culture.


CONCLUSIONS
After vocal fold injury, locally injected Carbylan-GSX can enhance the role of TNF-alpha in remodeling the lamina propria layer of the vocal fold, accelerating wound healing. Carbylan-GSX has potential as a new therapeutic approach that may lead to better treatment of vocal fold wound healing.
Chen
Xia
X
Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of Wisconsin Madison, Madison, Wisconsin 53705-2275, USA.
Thibeault
Susan L
SL
D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D024042:Collagen Type I / Q000032:analysis; D024043:Collagen Type II / Q000032:analysis; D005347:Fibroblasts / Q000187:drug effects / Q000502:physiology*; D005353:Fibronectins / Q000032:analysis; D005780:Gelatin / Q000494:pharmacology; D006801:Humans; D006820:Hyaluronic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D066298:In Vitro Techniques; D020781:Matrix Metalloproteinase 1 / Q000032:analysis; D020778:Matrix Metalloproteinase 2 / Q000032:analysis; D018124:Receptors, Tumor Necrosis Factor / Q000032:analysis; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000032:analysis; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology / Q000502:physiology*; D014827:Vocal Cords / Q000187:drug effects / Q000502:physiology*; D014945:Wound Healing / Q000187:drug effects / Q000502:physiology*
D024042:Collagen Type I; D024043:Collagen Type II; D005353:Fibronectins; D018124:Receptors, Tumor Necrosis Factor; D019717:Tissue Inhibitor of Metalloproteinase-3; D014409:Tumor Necrosis Factor-alpha; C513409:carbylan-GSX; D005780:Gelatin; D006820:Hyaluronic Acid; D020778:Matrix Metalloproteinase 2; D020781:Matrix Metalloproteinase 1