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Home / Equine Research Bank / Vet Ophthalmol / Efficacy and safety of mitomycin C as an agent to treat corn...
Veterinary ophthalmology2010; 13(4); 211-218; doi: 10.1111/j.1463-5224.2010.00782.x

Efficacy and safety of mitomycin C as an agent to treat corneal scarring in horses using an in vitro model.

Abstract: Mitomycin C (MMC) is used clinically to treat corneal scarring in human patients. We investigated the safety and efficacy of MMC to treat corneal scarring in horses by examining its effects at the early and late stages of disease using an in vitro model. Methods: An in vitro model of equine corneal fibroblast (ECF) developed was used. The ECF or myofibroblast cultures were produced by growing primary ECF in the presence or absence of transforming growth factor beta-1 (TGFbeta1) under serum-free conditions. The MMC dose for the equine cornea was defined with dose-dependent trypan blue exclusion and (3-4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays after applying MMC to the cultures once for 2 min. The efficacy of MMC to control corneal scarring in horses was determined by measuring mRNA and protein expression of corneal scarring markers (alpha-smooth muscle actin and F-actin) with western blotting, immunocytochemistry and/or quantitative real-time polymerase chain reactions. Results: A single 2-min treatment of 0.02% or less MMC did not alter ECF phenotype, viability, or cellular proliferation whereas 0.05% or higher MMC doses showed mild-to-moderate cellular toxicity. The TGFbeta1 at 1 ng/mL showed significant myofibroblast formation in ECF under serum-free conditions. A single 2-min, 0.02% MMC treatment 24 h (early) after TGFbeta1 stimulation significantly reduced conversion of ECF to myofibroblasts, however, a single 0.02% MMC treatment 11 days after TGFbeta1 stimulation showed moderate myofibroblast inhibition. Conclusions: That MMC safely and effectively reduced scarring in ECF by reducing the degree of transdifferentiation of corneal fibroblasts to myofibroblasts in vitro. Further clinical in vivo investigations are warranted using MMC in horses.
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Publication Date: 2010-07-14 PubMed ID: 20618797PubMed Central: PMC2904635DOI: 10.1111/j.1463-5224.2010.00782.xGoogle Scholar: Lookup
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  • Journal Article
  • Research Support
  • N.I.H.
  • Extramural
  • Research Support
  • Non-U.S. Gov't

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.

This research focuses on the use of Mitomycin C (MMC) to treat corneal scarring in horses, using an in vitro model. Experiments suggest that MMC can reduce the conversion of corneal fibroblasts to myofibroblasts, thus reducing scarring.

Methods

  • The researchers developed an in vitro model using equine corneal fibroblast (ECF).
  • ECF or myofibroblast cultures were grown in the presence or absence of a growth factor, TGFbeta1, in serum-free conditions to simulate the disease environment.
  • The appropriate dose of MMC for the equine cornea was established through trypan blue exclusion and MTT assays after a brief 2-minute exposure to MMC.
  • The efficacy of MMC in controlling corneal scarring was determined by measuring the expression of scarring markers at the molecular level (mRNA/proteins), with the use of specialized laboratory techniques like western blotting, immunocytochemistry, and polymerase chain reactions.

Results

  • Treatment with a dose of 0.02% or less MMC caused no significant changes in ECF phenotype, viability or cellular proliferation. In contrast, higher doses (0.05% or more) were mildly to moderately toxic to cells.
  • Exposure to 1 ng/mL of TGFbeta1 led to notable myofibroblast formation in ECF under serum-free conditions, mimicking disease conditions.
  • A single 2-minute treatment with 0.02% MMC, administered 24 hours after TGFbeta1 stimulation, significantly reduced the conversion of ECF to myofibroblasts. When the same treatment was applied 11 days post-TGFbeta1 stimulation, it showed moderate inhibition of myofibroblast formation.

Conclusions

  • The results indicate that MMC can safely and effectively reduce scarring by reducing the transdifferentiation of corneal fibroblasts into myofibroblasts in an in vitro setting.
  • Given the significance of these findings, the authors suggest that further clinical trials in live horses could establish the efficacy of MMC in treating corneal scarring.

Cite This Article

APA
Buss DG, Sharma A, Giuliano EA, Mohan RR. (2010). Efficacy and safety of mitomycin C as an agent to treat corneal scarring in horses using an in vitro model. Vet Ophthalmol, 13(4), 211-218. https://doi.org/10.1111/j.1463-5224.2010.00782.x

Publication

Veterinary ophthalmology
ISSN: 1463-5224
NlmUniqueID: 100887377
Country: England
Language: English
Volume: 13
Issue: 4
Pages: 211-218

Researcher Affiliations

Buss, Dylan G
  • Harry S. Truman Memorial Veterans Hospital, Columbia, MO, USA.
Sharma, Ajay
    Giuliano, Elizabeth A
      Mohan, Rajiv R

        MeSH Terms

        • Animals
        • Antibiotics, Antineoplastic / adverse effects
        • Antibiotics, Antineoplastic / therapeutic use
        • Apoptosis / drug effects
        • Blotting, Western / veterinary
        • Cells, Cultured
        • Cicatrix / drug therapy
        • Cicatrix / veterinary
        • Corneal Diseases / drug therapy
        • Corneal Diseases / veterinary
        • Dose-Response Relationship, Drug
        • Fibroblasts / drug effects
        • Horse Diseases / drug therapy
        • Horses
        • Immunoblotting / veterinary
        • In Vitro Techniques
        • Mitomycin / adverse effects
        • Mitomycin / therapeutic use
        • Polymerase Chain Reaction / veterinary

        Grant Funding

        • R01 EY017294 / NEI NIH HHS
        • I01 BX000357 / BLRD VA
        • R01 EY017294-02 / NEI NIH HHS
        • R01EY017294 / NEI NIH HHS
        • R01 EY017294-03S2 / NEI NIH HHS

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        Citations

        This article has been cited 8 times.
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