Related Experiment Videos
Effects of cyclosporin A on human conjunctival fibroblasts
A Leonardi1, G DeFranchis, I A Fregona
1Ophthalmology and Occular Inflammation Unit, Department of Neuroscience, University of Padova, Italy. mdvol@tin.it
Objective:
To evaluate the effects of cyclosporin A (CsA) on cytokine and/or collagen production, cell growth, and apoptosis in conjunctival fibroblast cultures.
Methods:
Fibroblast cultures derived from normal subjects and patients with vernal keratoconjunctivitis and pemphigoid were exposed to different concentrations of CsA for either 24 hours or 30 days. The effects were evaluated by the colorimetric MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) test to assess cell proliferation, and by the measurement of procollagen I (PIP) and procollagen III (PIIIP) cytokines and total protein in culture medium. CsA-induced apoptosis was assessed by fluorescence-activated cell sorter analysis.
Results:
After 24 hours of exposure to doses of CsA of more than 10 microg/mL, cell proliferation and migration were significantly reduced. Cyclosporin A reduced PIP and interleukin 1 (IL-1) production in a dose-dependent manner. Interleukin 6 and IL-8 were increased by 10 microg/mL of CsA, whereas transforming growth factor beta, PIIIP, and total protein were unaffected. Cyclosporin A exposure induced apoptosis in a time- and dose-dependent manner. Long-term exposure to CsA reduced IL-6 but did not modify PIIIP production.
Conclusion:
Exposure to CsA directly modified fibroblast behavior.
Clinical Relevance:
Cyclosporin A ability to accelerate apoptosis in clinically fibrotic tissues may prove to be therapeutic and useful in hyperproliferative conjunctival disorders.