Effects of Pirfenidone on Proliferation, Migration, and Collagen Contraction of Human Tenon's Fibroblasts In Vitro

To investigate the effect of pirfenidone, a novel antifibrotic agent, on proliferation, migration, and collagen contraction of human Tenon's fibroblasts (HTFs). After treatment of HTFs with pirfenidone, cell proliferation was measured by MTT assay. Cell migration was investigated by scratch ass...

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Bibliographic Details
Published in:Investigative ophthalmology & visual science 2009-08, Vol.50 (8), p.3763-3770
Main Authors: Lin, Xianchai, Yu, Minbin, Wu, Kaili, Yuan, Hongzhi, Zhong, Hua
Format: Article
Language:English
Subjects:
Anti-Inflammatory Agents, Non-Steroidal - pharmacology
Biological and medical sciences
Blotting, Western
Cell Cycle - drug effects
Cell Movement - drug effects
Cell Proliferation - drug effects
Cell Survival
Cells, Cultured
Collagen - metabolism
Dose-Response Relationship, Drug
Eye and associated structures. Visual pathways and centers. Vision
Fascia - cytology
Fibroblasts - cytology
Fibroblasts - drug effects
Fibroblasts - metabolism
Flow Cytometry
Fluorescent Antibody Technique, Indirect
Fundamental and applied biological sciences. Psychology
Humans
Medical sciences
Ophthalmology
Pyridones - pharmacology
Reverse Transcriptase Polymerase Chain Reaction
RNA, Messenger - metabolism
Transforming Growth Factor beta1 - genetics
Transforming Growth Factor beta1 - metabolism
Transforming Growth Factor beta2 - genetics
Transforming Growth Factor beta2 - metabolism
Transforming Growth Factor beta3 - genetics
Transforming Growth Factor beta3 - metabolism
Vertebrates: nervous system and sense organs
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Summary:To investigate the effect of pirfenidone, a novel antifibrotic agent, on proliferation, migration, and collagen contraction of human Tenon's fibroblasts (HTFs). After treatment of HTFs with pirfenidone, cell proliferation was measured by MTT assay. Cell migration was investigated by scratch assay. Contractility was evaluated in fibroblast-populated collagen gels. Cell viability was determined by trypan blue exclusion assay. The expression of TGF-beta1, -beta2, and -beta3 was estimated with RT-PCR, Western blot, and immunofluorescence analyses. Pirfenidone induced significant dose-dependent inhibition of HTF proliferation and migration and collagen contraction. After treatment with different concentrations of pirfenidone (0.15, 0.3, and 1 mg/mL) for 24 and 72 hours, cell viability was not different in the treatment and control groups. After 24 hours of treatment with pirfenidone, HTFs showed dose-dependent decreases in mRNA and protein levels of TGF-beta1, -beta2, and -beta3. These findings indicate that pirfenidone inhibits proliferation, migration, and collagen contraction of HTFs at nontoxic concentrations. A decrease in autocrine TGF-beta signaling may have a role in the effects of pirfenidone.
ISSN:0146-0404
1552-5783
1552-5783
DOI:10.1167/iovs.08-2815