Tocotrienol inhibits proliferation of human Tenon’s fibroblasts in vitro: a comparative study with vitamin E forms and mitomycin C

  • Alexander Meyenberg
  • David Goldblum
  • Jean-Marc Zingg
  • Angelo Azzi
  • Kalanithi Nesaretnam
  • Monika Kilchenmann
  • Beatrice E. Frueh
Laboratory Investigation



To evaluate the potential of the vitamin E compound α-tocotrienol as antifibrotic agent in vitro.


Using human Tenon’s capsule fibroblast cultures, the antiproliferative and cytotoxic effects of the different vitamin E forms α-tocopherol, α-tocopheryl acetate, α-tocopheryl succinate and α-tocotrienol were compared with those of mitomycin C. To mimic subconjunctival and regular oral application in vivo, exposure time of serum-stimulated and serum-restimulated fibroblasts (SF and RF, respectively) to vitamin E forms was set at 6 days. Cultures were only exposed for 5 min to mitomycin C due to its known acute toxicity and to mimic the short-time intraoperative administration. Proliferation (expressed as % of control) was determined by DNA content quantification on days 2, 4 and 6, whereas cytotoxicity was assessed by cell morphology and glucose 6-phosphate dehydrogenase (G6PD) release after 24 h.


α-Tocopherol and α-tocopheryl acetate stimulated growth of SF, but not RF. Reduction of fibroblast content by α-tocopheryl succinate was accompanied by increased G6PD release and necrosis. Contrary to α-tocopheryl succinate, 50 μM or repeatedly 20 μM of α-tocotrienol significantly inhibited proliferation without causing cellular toxicity (maximal effect: 46.8%). RF were more sensitive to this effect than SF. Mitomycin C 100–400 μg/ml showed a stronger antiproliferative effect than α-tocotrienol (maximal effect: 13.8%). Morphologic characteristics of apoptosis were more commonly found under treatment with mitomycin C.


Of the vitamin E forms tested, only α-tocotrienol significantly inhibited growth at non-toxic concentrations. In this in vitro study, antiproliferative effects of mitomycin C were stronger than those of α-tocotrienol.


Total Cell Lysis Tocotrienols Degenerative Cell Change 



The authors thank Milko E. Iliev, MD, for helpful discussions and critical comments on the manuscript.

Financial interests: A. Meyenberg, none; D. Goldblum, none; J.-M. Zingg, none; A. Azzi, none; K. Nesaretnam, employee of the Malaysian Palm Oil Board, no proprietary interests; M. Kilchenmann, none; B. E. Frueh, none.

The authors have full control of all primary data and they agree to allow Graefe’s Archive for Clinical and Experimental Ophthalmology to review their data if requested.


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Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Alexander Meyenberg
    • 1
  • David Goldblum
    • 1
  • Jean-Marc Zingg
    • 2
  • Angelo Azzi
    • 2
  • Kalanithi Nesaretnam
    • 3
  • Monika Kilchenmann
    • 1
  • Beatrice E. Frueh
    • 1
  1. 1.Departement of OphthalmologyUniversity of Berne, Inselspital BerneBerneSwitzerland
  2. 2.Institute for Biochemistry and Molecular BiologyUniversity of BerneBerneSwitzerland
  3. 3.Malaysian Palm Oil BoardKuala LumpurMalaysia

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