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1 From the Institute of Wound Research, Department of Obstetrics and Gynecology and the 2 Department of Ophthalmology, University of Florida, Gainesville; and the 3 Department of Ophthalmology, University of South Florida, Tampa.
PURPOSE. To assess the role of the transforming growth factor (TGF)ß system in formation of corneal haze after excimer laser photorefractive keratectomy (PRK), levels of mRNAs for three TGFß isoforms (TGFß1, TGFß2, and TGFß3), the TGFß type II receptor (TßRII), and extracellular matrix (ECM) genes including fibronectin (FN), collagen I, collagen III, and collagen IV were measured in rat corneas.
METHODS. Corneas were graded for corneal haze at 0, 1.5, 7, 21, 42, and 91 days after PRK. Total RNA was isolated from pooled corneas, and the levels of mRNAs were measured using competition-based quantitative reverse transcription–polymerase chain reaction (RT-PCR).
RESULTS. Severe corneal haze developed by day 42 and persisted to day 91. Levels
of TGFß1 mRNA were high in rat corneas before PRK and remained
relatively constant. In contrast, levels of TGFß2 and TGFß3 mRNAs
were very low in normal corneas, increased 300-fold and 25-fold,
respectively, on day 21, and remained elevated on day 91. Levels of
mRNA for TßRII increased, with a peak elevation of 50-fold on day 42
after PRK. Levels of mRNAs for ECM proteins also increased. Fibronectin
mRNA was nondetectable in normal corneas but rapidly increased to 675
copies/cell on day 7 and remained elevated to day 91. Collagen III mRNA
levels peaked on day 21 with a 700-fold increase compared with a very
low level of expression in normal cornea, and then decreased on day 91.
Expression of collagen I mRNA lagged expression of collagen III mRNA
and peaked at day 42 after PRK with a 1200-fold increase over normal
cornea. In contrast, mRNA for collagen 
(1)IV, a major component in
basement membranes, remained relatively stable through day 21 and then
increased slightly on days 42 and 91.
CONCLUSIONS. The synchronized increase in mRNA synthesis for both the TGFß system and key ECM genes supports the hypothesis that TGFß is a key growth factor promoting stromal haze formation in corneas after PRK and suggests that limiting TGFß system may reduce corneal scarring after excimer laser ablation.
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