Anti-fibrotic action of pirfenidone in Dupuytren's disease-derived fibroblasts.
Dupuytren's disease (DD) is a complex fibro-proliferative disorder of the hand that is often progressive and eventually can cause contractures of the affected fingers. Transforming growth factor beta (TGF-β1) has been implicated as a key stimulator of myofibroblast activity and fascial contraction in DD. Pirfenidone (PFD) is an active small molecule shown to inhibit TGF-β1-mediated action in other fibrotic disorders. This study investigates the efficacy of PFD in vitro in inhibiting TGF-β1-mediated cellular functions leading to Dupuytren's fibrosis.



Fibroblasts harvested from (DD) and carpal tunnel (CT)- tissues were treated with or without TGF-β1and/or PFD and were subjected to cell migration, cell proliferation and cell contraction assays. ELISA; western blots and real time RT-PCR assays were performed to determine the levels of fibronectin; p-Smad2/Smad3; alpha-smooth muscle actin (α-SMA), α2 chain of type I collagen and α1 chain of type III collagen respectively.



Our results show that PFD effectively inhibits TGF-β1-induced cell migration, proliferation and cell contractile properties of both CT- and DD-derived fibroblasts. TGF-β1-induced α-SMA mRNA and protein levels were inhibited at the higher concentration of PFD (800 μg/ml). Interestingly, TGF-β1induction of type I and type III collagens and fibronectin was inhibited by PFD in both CT- and DD- derived fibroblasts, but the effect was more prominent in DD cells. PFD down-regulated TGF-β1-induced phosphorylation of Smad2/Smad3, a key factor in the TGF-β1signaling pathway.



Taken together these results suggest the PFD can potentially prevent TGF-β1-induced fibroblast to myofibroblast transformation and inhibit ECM production mainly Type I- and Type III- collagen and fibronectin in DD-derived fibroblasts. Further in-vivo studies with PFD may lead to a novel therapeutic application in preventing the progression or recurrence of Dupuytren's disease.
Department of Plastic Surgery, University of Pittsburgh, Pittsburgh, PA, 15261, USA.
Chaoming
C
Zhou
Department of Plastic Surgery, University of Pittsburgh, Pittsburgh, PA, 15261, USA.
Fang
F
Liu
Department of Plastic Surgery, University of Pittsburgh, Pittsburgh, PA, 15261, USA.
Phillip H
PH
Gallo
Department of Orthopaedic Surgery, University of Pittsburgh, Pittsburgh, PA, 15261, USA.
Mark E
ME
Baratz
McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, USA.
Sandeep
S
Kathju
McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, USA. las238@pitt.edu.|Department of Plastic Surgery, University of Pittsburgh, 3550 Terrace Street, Scaife Hall, S685.2, Pittsburgh, PA, 15261, USA. las238@pitt.edu.
Latha
L
Satish
D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology / Q000627:therapeutic use*; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D024042:Collagen Type I / Q000378:metabolism; D024061:Collagen Type III / Q000378:metabolism; D004353:Drug Evaluation, Preclinical; D004387:Dupuytren Contracture / Q000188:drug therapy*; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D006801:Humans; D010766:Phosphorylation / Q000187:drug effects; D061251:Primary Cell Culture; D011728:Pyridones / Q000494:pharmacology / Q000627:therapeutic use*; D051899:Smad2 Protein / Q000378:metabolism; D051900:Smad3 Protein / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000378:metabolism
D000894:Anti-Inflammatory Agents, Non-Steroidal; D024042:Collagen Type I; D024061:Collagen Type III; D005353:Fibronectins; D011728:Pyridones; C497782:SMAD2 protein, human; C497790:SMAD3 protein, human; D051899:Smad2 Protein; D051900:Smad3 Protein; C508960:TGFB1 protein, human; D053773:Transforming Growth Factor beta1; C093844:pirfenidone
Alpha-SMA; Carpal tunnel; Cell contraction; Cell migration; Collagen; Dupuytren’s contracture; Palmar fascia fibrosis; Smad2/Smad3