Vocal fold myofibroblast profile of scarring

Abstract

Objectives/Hypothesis Vocal fold fibroblasts (VFF) are responsible for extracellular matrix synthesis supporting lamina propria in normal and diseased conditions. When tissue is injured, VFF become activated and differentiate into myofibroblasts to facilitate wound healing response. We investigated if vocal fold myofibroblasts can be utilized as surrogate cells for scarred VFF. Study Design In vitro. Methods Normal VFF cell lines from a 21-year-old male (N21), 59-year-old female (N59), and a scar VFF cell line from a 56 year-old female (S56) were used in this study. 10ng/mL TGFβ1 was applied for 5 days to normal VFF. Myofibroblast differentiation was determined with immunocytochemistry and western blot, measuring α-SMA. Cell growth, proliferation, contractile properties and gene expression profiles were evaluated. Results N21, N59 and S56 VFF presented elongated configuration. N21+ and N21− VFF demonstrated significantly greater proliferation compared to N59+, N59− and S56 VFF at 6 days. α-SMA was expressed in all cells. Fibronectin, α-SMA, CTGF and TIMP were the highest genes expression in VFF treated with TGFβ1. At 24 hours, S56 VFF showed lower contraction compared to N21+ and N59+ VFF, but at 60 hours, S56 VFF had lower collagen contraction compared to all cell groups. Highest collagen contraction matrices were measured with VFF treated with TGFβ1 at 24 hours and N59− VFF at 60 hours. Conclusion VFF treated with TGFβ1 (myofibroblasts) appear to have similar phenotypic characteristics but different genotypic behavior compared to scar VFF.

Photodynamic therapy induces antifibrotic alterations in primary human vocal fold fibroblasts

“…However, excessive activation of fibroblasts into myofibroblasts can result in fibrosis . α‐SMA protein is the marker of myofibroblasts and one of the proteins responsible for scar contraction . In our study, TGF‐β1 treatment for 5 days significantly upregulated the expression of α‐SMA; this expression was significantly decreased after SL‐PDT.…”

“…Recombinant human TGF‐β1 was purchased from Peprotech (100‐21C; Peprotech, Rocky Hill, NJ). To induce the transition from fibroblasts to myofibroblasts, VFFs received 5 days of treatment of DMEM + 2.5% FBS and 10 ng/mL TGF‐β1 . The medium was removed and cells were rinsed with PBS three times.…”

“…However, these effects have yet to be determined in vocal fold fibroblasts (VFFs), which is the main cell population in vocal fold lamina propria. VFFs play an important role in the formation and maintenance of ECM by producing, for example, collagen, elastin, fibronectin, matrix metallopeptidases (MMPs), and TGF‐β1 . No study has been performed to investigate the effects of PDT on VFFs and vocal fold ECM formation.…”

Cryotherapy has antifibrotic and regenerative effects on human vocal fold fibroblasts

“…Vocal fold scarring continues to pose a significant clinical challenge, which is characterized by disordered extracellular matrix (ECM) in the lamina propria (LP). This is typically recognized by increased collagen deposition, poorly organized collagen orientation, and decreased proteoglycan density . Vocal fold scarring will reduce voice quality and can even result in a significant dysphonia that is difficult to treat .…”

“…When attack happens on the vocal folds, VFFs may exhibit an unbalanced production of ECM components, and the most extreme response to injury is scarring. One of the early steps in wound healing involves the recruitment of fibroblasts to the site of injury . Therefore, two issues must be addressed for preventing vocal fold scarring: 1) the aberrant synthetic phenotype of the VFFs after injury and 2) the regeneration of the LP with organization and orientation of novel collagen fibers.…”