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Updated: Jun 26, 2026
Ex Vivo Corneal Organ Culture Model for Wound Healing Studies
Published on: February 15, 2019
Trichostatin a inhibits corneal haze in vitro and in vivo
Ajay Sharma1, Maneesh M Mehan, Sunilima Sinha
1Mason Eye Institute, University of Missouri-Columbia, Columbia, Missouri 65212, USA.
Purpose:
Trichostatin A (TSA), a histone deacetylase inhibitor, has been shown to suppress TGF-beta-induced fibrogenesis in many nonocular tissues. The authors evaluated TSA cytotoxicity and its antifibrogenic activity on TGF-beta-driven fibrosis in the cornea with the use of in vitro and in vivo models.
Methods:
Human corneal fibroblasts (HSFs) were used for in vitro studies, and New Zealand White rabbits were used for in vivo studies. Haze in the rabbit cornea was produced with photorefractive keratectomy (PRK) using excimer laser. Trypan blue exclusion and MTT assays evaluated TSA cytotoxicity to the cornea. Density of haze in the rabbit eye was graded with slit lamp biomicroscopy. Real-time PCR, immunoblotting, or immunocytochemistry was used to measure alpha-smooth muscle actin (SMA), fibronectin, and collagen type IV mRNA or protein levels. TUNEL assay was used to detect cell death.
Results:
TSA concentrations of 250 nM or less were noncytotoxic and did not alter normal HSF morphology or proliferation. TGF-beta1 treatment of HSF significantly increased mRNA and protein levels of SMA (9-fold), fibronectin (2.5-fold), and collagen type IV (2-fold). TSA treatment showed 60% to 75% decreases in TGF-beta1-induced SMA and fibronectin mRNA levels and 1.5- to 3.0-fold decreases in protein levels but had no effect on collagen type IV mRNA or protein levels in vitro. Two-minute topical treatment of TSA on rabbit corneas subjected to -9 D PRK significantly decreased corneal haze in vivo.
Conclusions:
TSA inhibits TGF-beta1-induced accumulation of extracellular matrix and myofibroblast formation in the human cornea in vitro and markedly decreases haze in rabbit cornea in vivo.
Insights
Trichostatin A (TSA) effectively reduces corneal fibrosis and haze by inhibiting extracellular matrix accumulation and myofibroblast formation. This histone deacetylase inhibitor shows promise for treating TGF-beta-driven corneal scarring in vivo and in vitro.
Area of Science:
- Ophthalmology
- Cell Biology
- Pharmacology
Background:
- Corneal fibrosis, a leading cause of blindness, is often driven by transforming growth factor-beta (TGF-beta).
- Histone deacetylase inhibitors, such as Trichostatin A (TSA), have demonstrated antifibrotic effects in nonocular tissues.
- The potential of TSA to treat corneal fibrosis remains largely unexplored.
Purpose of the Study:
- To evaluate the cytotoxicity of TSA on corneal cells.
- To assess the antifibrogenic activity of TSA in TGF-beta-driven corneal fibrosis models.
- To investigate the efficacy of TSA in reducing corneal haze in vivo.
Main Methods:
- In vitro studies utilized human corneal fibroblasts (HSFs) treated with TGF-beta1 and TSA.
- In vivo studies involved New Zealand White rabbits undergoing photorefractive keratectomy (PRK) followed by topical TSA treatment.
- Cytotoxicity was assessed using Trypan blue exclusion and MTT assays. Fibrosis markers (alpha-SMA, fibronectin, collagen IV) and corneal haze were quantified.
Main Results:
- TSA at concentrations up to 250 nM was noncytotoxic to HSFs.
- TSA significantly reduced TGF-beta1-induced alpha-smooth muscle actin (SMA) and fibronectin expression in HSFs.
- Topical TSA treatment markedly decreased corneal haze in rabbits post-PRK.
Conclusions:
- TSA effectively inhibits TGF-beta1-induced extracellular matrix accumulation and myofibroblast differentiation in corneal fibroblasts.
- TSA demonstrates significant efficacy in reducing corneal haze in a rabbit model of photorefractive keratectomy.
- TSA represents a potential therapeutic agent for managing corneal fibrosis and scarring.
