July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Transient mitomycin C treatment of human corneal fibroblasts reduces collagen deposition and secretion of CCL2 and CXCL1.
Author Affiliations & Notes
  • Sonali Ghosh
    Anatomy and Cell Biology, GWU Medical School, Washington, District of Columbia, United States
  • Gauri Tadvalkar
    Anatomy and Cell Biology, GWU Medical School, Washington, District of Columbia, United States
  • Audrey E K Hutcheon
    Ophthalmology, Schepen's Eye Research Institute Harvard, Boston, Massachusetts, United States
  • Xiaoqing Q Guo
    Ophthalmology, Schepen's Eye Research Institute Harvard, Boston, Massachusetts, United States
  • James D. Zieske
    Ophthalmology, Schepen's Eye Research Institute Harvard, Boston, Massachusetts, United States
  • Mary Ann Stepp
    Anatomy and Cell Biology, GWU Medical School, Washington, District of Columbia, United States
  • Footnotes
    Commercial Relationships   Sonali Ghosh, None; Gauri Tadvalkar, None; Audrey Hutcheon, None; Xiaoqing Guo, None; James Zieske, None; Mary Ann Stepp, None
  • Footnotes
    Support  NIH/NEI EY08512 and EY021784 (MAS)
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 4660. doi:
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      Sonali Ghosh, Gauri Tadvalkar, Audrey E K Hutcheon, Xiaoqing Q Guo, James D. Zieske, Mary Ann Stepp; Transient mitomycin C treatment of human corneal fibroblasts reduces collagen deposition and secretion of CCL2 and CXCL1.. Invest. Ophthalmol. Vis. Sci. 2019;60(9):4660.

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      © ARVO (1962-2015); The Authors (2016-present)

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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.

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