May 1995
Volume 36, Issue 6
Free
Articles  |   May 1995
Effect of tecogalan sodium on angiogenesis in vitro by choroidal endothelial cells.
Author Affiliations
  • T Sakamoto
    Doheny Eye Institute, University of Southern California School of Medicine, Los Angeles 90033, USA.
  • T Ishibashi
    Doheny Eye Institute, University of Southern California School of Medicine, Los Angeles 90033, USA.
  • H Kimura
    Doheny Eye Institute, University of Southern California School of Medicine, Los Angeles 90033, USA.
  • H Yoshikawa
    Doheny Eye Institute, University of Southern California School of Medicine, Los Angeles 90033, USA.
  • C Spee
    Doheny Eye Institute, University of Southern California School of Medicine, Los Angeles 90033, USA.
  • M S Harris
    Doheny Eye Institute, University of Southern California School of Medicine, Los Angeles 90033, USA.
  • D R Hinton
    Doheny Eye Institute, University of Southern California School of Medicine, Los Angeles 90033, USA.
  • S J Ryan
    Doheny Eye Institute, University of Southern California School of Medicine, Los Angeles 90033, USA.
Investigative Ophthalmology & Visual Science May 1995, Vol.36, 1076-1083. doi:
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    • Get Citation

      T Sakamoto, T Ishibashi, H Kimura, H Yoshikawa, C Spee, M S Harris, D R Hinton, S J Ryan; Effect of tecogalan sodium on angiogenesis in vitro by choroidal endothelial cells.. Invest. Ophthalmol. Vis. Sci. 1995;36(6):1076-1083.

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

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Abstract

PURPOSE: To examine the possible inhibitory effect of tecogalan sodium, derived from bacteria, on three important components of in vitro angiogenesis (endothelial proliferation, migration, and tube formation in a collagen gel) using bovine choroidal endothelial cells (CECs). METHODS: The effects of tecogalan sodium (1, 5, 25, 125, and 250 micrograms/ml) on cultured CECs were examined when basic fibroblast growth factor (bFGF, 10 ng/ml), vascular endothelial growth factor (VEGF, 50 ng/ml), a combination of bFGF (10 ng/ml) and VEGF (50 ng/ml) (bFGF/VEGF) and 10% fetal calf serum (FCS) were used as angiogenic stimulants. For the proliferation assay, CECs were cultured and the cell numbers counted on days 1, 3, and 5. For migration assay, CECs were seeded in the upper half of a Boyden chamber while an angiogenic growth factor was loaded in the lower half. After 6 hours of incubation, cell migration was evaluated by counting the numbers of migrated cells per microscopic field on the lower side of the filter. For the tube-forming assay, CECs were seeded in a type I collagen gel, and the length of the tube-like structures (an indicator of angiogenesis) formed by CECs per microscopic field was quantified by image analysis. The effect of neutralizing antibody for bFGF also was tested in these three assays. RESULTS: All tested angiogenic stimulants induced CEC proliferation. The stimulatory effect of bFGF and bFGF/VEGF was reduced by tecogalan sodium (IC50 for bFGF effect, 26.1 micrograms/ml). However, the effect of VEGF and of 10% FCS was not altered by low doses of tecogalan sodium (< 25 micrograms/ml). Chemotaxis of CECs was stimulated by bFGF alone and by bFGF/VEGF, and this effect was inhibited by tecogalan sodium (IC50 for bFGF, 3.2 micrograms/ml). Stimulation of chemotaxis by VEGF alone and by 10% FCS was not affected by tecogalan sodium in low doses but was inhibited by high doses. Tube formation was stimulated by administration of each of the factors. Stimulation of tube formation by bFGF and by bFGF/VEGF was inhibited by tecogalan sodium (IC50 for bFGF, 18.2 micrograms/ml). High doses of tecogalan sodium (125 and 250 micrograms/ml) also inhibited 10% FCS-induced proliferation, migration, and tube formation. CONCLUSION: bFGF, VEGF, and a combination of bFGF and VEGF stimulated proliferation, migration, and tube formation by CECs in vitro. These stimulatory effects, but especially those of bFGF, were inhibited by tecogalan sodium. If tecogalan sodium can be shown to have a similar effect in vivo, it might have the potential for pharmacologic control of subretinal neovascularization.

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