May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Human Serum reduces Mitomycin–C Cytotoxicity in Tenon’s Fibroblasts
X. Wang; J.G. Crowston; H. Zoellner; P.R. Healey
Author Affiliations & Notes
  • X. Wang
    Centre for Vision Research, Westmead Millennium Institute,
    University of Sydney, Sydney, Australia
  • J.G. Crowston
    Centre for Vision Research, Westmead Millennium Institute,
    University of Sydney, Sydney, Australia
  • H. Zoellner
    Department of Oral Pathology and Oral Medicine, Westmead Hospital,
    University of Sydney, Sydney, Australia
  • P.R. Healey
    Centre for Vision Research, Westmead Millennium Institute,
    University of Sydney, Sydney, Australia
  • Footnotes
    Commercial Relationships  X. Wang, None; J.G. Crowston, None; H. Zoellner, None; P.R. Healey, None.
  • Footnotes
    Support  none
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 927. doi:
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      X. Wang, J.G. Crowston, H. Zoellner, P.R. Healey; Human Serum reduces Mitomycin–C Cytotoxicity in Tenon’s Fibroblasts . Invest. Ophthalmol. Vis. Sci. 2004;45(13):927.

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Abstract

Abstract: : Purpose: Subconjunctival scarring, mediated by Tenon’s fibroblasts, is an important cause of trabeculectomy failure. Mitomycin–C (MMC) reduces fibroblast numbers by inhibiting proliferation and inducing apoptosis. The purpose of this study was to investigate the effects of human serum on the cytotoxic action of MMC in an in–vitro human Tenon’s fibroblast model. Methods: Cultured human Tenon’s fibroblasts were treated with MMC (0.4 mg/ml) in PBS for 3–5 minutes. Controls were treated with PBS only. After treatment, the cells were cultured in RPMI in the presence or absence of heat–inactivated human serum at varying concentrations. Cell number was measured by MTT assay, and cell death was determined by flow cytometry using Annexin V/propidium iodide cell labelling. Results: MMC reduced fibroblast numbers by 33% and 72% on days 3 and 7 in serum–free culture when compared with controls (P < 0.01, P<0.05). Exposure of cells to 10% of human serum resulted in a 2.3 fold increase in viable cell number on day 3 (P< 0.001) and a 3.3 fold increase on day 7 (P< 0.001) respectively. MMC caused 4.7–fold increase in cell death after 48 hours in serum–free culture. This was decreased by 32% in the presence of 10 % human serum. Conclusions:Human serum reduced the cytotoxic effect of MMC in Tenon’s fibroblast. This action is associated with a reduction in cell death.

Keywords: wound healing • cell death/apoptosis • blood supply 
© 2004, The Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Permission to republish any abstract or part of an abstract in any form must be obtained in writing from the ARVO Office prior to publication.
Copyright © 2025 Association for Research in Vision and Ophthalmology.
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