Transient Mitomycin C-treatment of human corneal epithelial cells and fibroblasts alters cell migration, cytokine secretion, and matrix accumulation

Abstract

A single application of Mitomycin C (MMC) is used clinically in ophthalmology to reduce scarring and enhance wound resolution after surgery. Here we show in vitro that a 3-hour MMC treatment of primary and telomerase immortalized human corneal limbal epithelial (HCLE) cells impacts their migration and adhesion. Transient MMC treatment induces HCLE expression of senescence associated secretory factors, cytokine secretion, and deposition of laminin 332 for several days. Transient MMC treatment also reduces migration and deposition of transforming growth factor-β1 (TGFβ1)-stimulated collagen by corneal fibroblasts. Using conditioned media from control and MMC treated cells, we demonstrate that factors secreted by MMC-treated corneal epithelial cells attenuate collagen deposition by HCFs whereas those secreted by MMC-treated HCFs do not. These studies are the first to probe the roles played by corneal epithelial cells in reducing collagen deposition by corneal fibroblasts in response to MMC.

Safety and efficacy of combination of suberoylamilide hydroxyamic acid and mitomycin C in reducing pro-fibrotic changes in human corneal epithelial cells

“…MMC prevents sub-epithelial haze by inducing senescence in activated keratocytes thereby reducing the abnormal ECM by attenuating the TGFβ1 signalling in activated fibroblasts 8 , 31 . Epithelial basement membrane proteins such as laminin332, perlecan, nidogen are enhanced by MMC treated corneal epithelial cells 9 , 32 . These proteins play vital role in preventing haze.…”

“…The proliferation and migration of the activated keratocytes in the stromal region gets initiated 12–14 h post corneal epithelial injury activated by surgery 6 . However, studies have shown that the status of corneal epithelial cells also contributes in triggering the cascade leading to haze resolution 9 . Further the effect of drugs to treat haze on limbus need to be addressed as long term outcome may lead to limbal stem cell deficiency.…”

“…Animal studies have shown protective effect of SAHA on endothelial cells 26 . It has been illustrated that senescence associated secretory proteins from MMC treated corneal epithelial cells play pivotal role in mitigating the role of MMC in preventing haze 9 . Usage of MMC in ophthalmic surgery was first done as an adjunct in pterygium excision in 1963 and it first use as a corneal healing modulator was performed after almost after more than 35 years in PRK 29 , 30 .…”

“…The cause of haze is being triggered by the initial surgery induced insult to corneal epithelial cells. However, further studies have revealed that MMC reduces corneal fibrosis by reducing collagen deposition and expression of αSMA and fibronectin by corneal fibroblasts and simultaneous deposition of epithelial basement membrane protein Laminin332 9 .…”

“…It has been approved to be used for preventing haze on corneal surface by inducing apoptosis of keratocytes and stromal cells 8 . It has been elucidated that conditioned medium from MMC treated corneal epithelial cells lead to senescence of corneal epithelial cells and secreted senescence associated secreted proteins that suppressed the collagen deposition implicating a definitive role of corneal epithelial cells in preventing haze in the presence of MMC 9 . It has been shown that prophylactic usage of MMC has more beneficial effects rather than therapeutic effects 10 .…”