Abstract
Purpose :
To determine in vitro the mechanism underlying the ability of mitomycin C to reduce stromal scar formation in vivo.
Methods :
Human corneal fibroblasts were obtained from human corneas and expanded and used within 3-4 passages. Confluent human corneal fibroblasts were treated with 0.02% MMC for 3 hours; cells were washed, media removed, and cells allowed to condition serum free media for 48 hr. Conditioned media from equal numbers of control or mitomycin C treated human corneal fibroblasts was used in cytokine array studies which were repeated twice. Human corneal fibroblasts were also treated with mitomycin C for 3 hours, washed, and grown in media supplemented with vitamin C and TGFb1 for 48 hrs. Collagen deposition and cell counts were assessed using picosirius red and crystal violet.
Results :
Cytokine arrays show significantly reduced expression of CCL2 and CXCL1 but no change in expression of MIF, PAI1/serpine1, and IL8 in mitomycin C treated conditioned media compared to controls. None of the cytokines assessed were increased in expression. While human corneal fibroblasts treated with vitamin C and TGFb1 increase collagen deposition compared to controls, mitomycin C-treated human corneal fibroblasts do not.
Conclusions :
These data show that mitomycin C impairs the ability of vitamin C and TGFb1 to upregulate collagen deposition in vitro by human corneal fibroblasts via a mechanism involving downregulation of corneal fibroblast secretion of CCL2 and CXCL1.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.