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Home / Equine Research Bank / Vet Ophthalmol / Therapeutic potential of Pirfenidone for treating equine cor...
Veterinary ophthalmology2014; 18(3); 242-250; doi: 10.1111/vop.12194

Therapeutic potential of Pirfenidone for treating equine corneal scarring.

Abstract: To evaluate the safety and efficacy of Pirfenidone (PFD) in the treatment of equine corneal fibrosis using an in vitro model. Methods: Healthy donor equine corneas were collected and used to generate primary equine corneal fibroblasts (ECFs) by growing cultures in minimal essential medium supplemented with 10% fetal bovine serum. Equine corneal myofibroblasts (ECMs), used as a model of equine corneal fibrosis, were produced by growing ECF cultures in serum-free medium containing transforming growth factor β1 (1 ng/mL). Trypan blue viability assays and changes in ECF morphology were utilized to determine the optimal PFD dose for this in vitro model. Trypan blue viability, phase-contrast microscopy, and TUNEL assays were used to evaluate the cytotoxicity of PFD. Scratch and MTT assays were used to evaluate the effect of PFD on cellular migration and proliferation. Real-time PCR, immunoblot analysis, and immunocytochemistry were employed to determine the efficacy of PFD to inhibit ECM formation in vitro. Results: Topical PFD application at 200 μg/mL successfully decreased αSMA expression when compared to the TGFβ1 only treatment group (P < 0.01). PFD application ≤ 200 μg/mL did not affect ECF phenotype or cellular viability and did not result in significant cytotoxicity. Conclusions: Pirfenidone safely and effectively inhibits TGFβ1-induced equine corneal fibrosis in vitro. In vivo studies are warranted.
© 2014 American College of Veterinary Ophthalmologists.
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Publication Date: 2014-07-15 PubMed ID: 25041235PubMed Central: PMC4295017DOI: 10.1111/vop.12194Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • N.I.H.
  • Extramural
  • Research Support
  • Non-U.S. Gov't
  • Research Support
  • U.S. Gov't
  • Non-P.H.S.

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article discusses an examination of the drug Pirfenidone’s potential in treating horse corneal scarring, experimenting with in vitro models.

Study Methodology

  • The study started by using healthy corneas from horses to produce primary equine corneal fibroblasts (ECFs), required in the generation of tissue and wound healing.
  • These ECFs were grown in a minimal essential medium containing 10% fetal bovine serum, provoked to transform into equine corneal myofibroblasts (ECMs), acting as a model for horse corneal fibrosis, using a serum-free medium with transforming growth factor β1 (TGFβ1).
  • To determine the optimal dosage of Pirfenidone for the model, the researchers observed changes in ECF morphology and used Trypan blue viability assays, an assay used to determine cell viability.
  • The possible cytotoxicity of the drug was evaluated using TUNEL assays, Trypan blue viability, and phase-contrast microscopy, a technique allowing detailed examination of cell structure.
  • The effect of Pirfenidone on cellular migration and proliferation was accessed using MTT and Scratch assays.
  • Real-time PCR, immunoblot analysis, and immunocytochemistry – advanced techniques for protein and genetic analysis – were used to ascertain if PFD could inhibit ECM formation.

Study Findings

  • The results displayed that a topical application of Pirfenidone at 200 μg/mL significantly reduced the expression of αSMA, a protein marker for myofibroblast development, when compared to a group treated only with TGFβ1.
  • Pirfenidone’s application did not affect ECF’s phenotype or their viability and was not observed to be significantly cytotoxic, deeming it a safe treatment option.

Conclusion

  • The study concludes that Pirfenidone demonstrated an ability to effectively and safely inhibit TGFβ1-induced equine corneal fibrosis in test tube models.
  • While the obtained results indicate promise, in vivo studies, where the drug is tested on living organisms, are recommended for further validations.

Cite This Article

APA
Fink MK, Giuliano EA, Tandon A, Mohan RR. (2014). Therapeutic potential of Pirfenidone for treating equine corneal scarring. Vet Ophthalmol, 18(3), 242-250. https://doi.org/10.1111/vop.12194

Publication

Veterinary ophthalmology
ISSN: 1463-5224
NlmUniqueID: 100887377
Country: England
Language: English
Volume: 18
Issue: 3
Pages: 242-250

Researcher Affiliations

Fink, Michael K
  • Harry S. Truman Memorial Veterans Hospital, Columbia, MO, USA; College of Veterinary Medicine, University of Missouri, Columbia, MO, USA.
Giuliano, Elizabeth A
    Tandon, Ashish
      Mohan, Rajiv R

        MeSH Terms

        • Actins / genetics
        • Actins / metabolism
        • Animals
        • Anti-Inflammatory Agents, Non-Steroidal / therapeutic use
        • Cell Movement / drug effects
        • Cells, Cultured
        • Cornea / cytology
        • Corneal Injuries / drug therapy
        • Corneal Injuries / veterinary
        • Fibroblasts / drug effects
        • Fibroblasts / physiology
        • Gene Expression Regulation / drug effects
        • Horse Diseases / drug therapy
        • Horses
        • Pyridones / therapeutic use

        Grant Funding

        • I01 BX000357 / BLRD VA
        • R01 EY017294 / NEI NIH HHS
        • R01EY017294 / NEI NIH HHS

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        Citations

        This article has been cited 14 times.
        1. Sinha NR, Hofmann AC, Suleiman LA, Jeffrey MT, Kumar R, Mohan RR. Alterations in Mitochondrial DNA in Corneal Fibroblast and Myofibroblast Post Injury. Invest Ophthalmol Vis Sci 2026 Jan 5;67(1):36.
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        3. Routh BL, Tripathi R, Giuliano E, Lujin P, Sinha PR, Mohan RR. Anti-fibrotic effects of lisinopril (ACE inhibitor) and fasudil (ROCK inhibitor) in combination for canine corneal fibrosis in vitro. Vet Ophthalmol 2026 Jan;29(1):e13304.
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