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Human Reproduction, Vol. 16, No. 7, 1457-1463, July 2001
© 2001 European Society of Human Reproduction and Embryology

Embryotoxicity of magainin-2-amide and its enhancement by cyclodextrin, albumin, hydrogen peroxide and acidification

Ewa T. Mystkowska1, Anna Niemierko1, Aldona Komar1 and Wojciech Sawicki2,3

1 Department of Histology and Embryology and 2 Department of Electron Microscopy, Center of Biostructure, Medical Academy, Warsaw, Poland

BACKGROUND: The channel-forming antimicrobial peptide, magainin-2-amide, interacts preferentially with negatively charged, non cholesterol-containing membranes, including those of sperm, oocytes and cells of pre-implantation embryos. Cyclodextrin and albumin remove membrane cholesterol and together with hydrogen peroxide (H2O2) are potential enhancers of embryotoxicity. METHODS: Two-cell murine embryos were cultured in vitro with magainin-2-amide at a high effective concentration (250 µg/ml) and at subthreshold concentrations (166 and 200 µg/ml). Embryos treated with sub-threshold concentrations of magainin were additionally treated with cyclodextrin, bovine serum albumin or H2O2 or were cultured under acidified conditions. Cell viability was verified with propidium iodide and fluorescein diacetate. RESULTS: The embryotoxic effect of magainin and H2O2 was dose- and time-dependent. Cyclodextrin, H2O2, acidification of the medium, and to a lesser extent albumin, enhanced the embryotoxicity of magainin at sub-threshold concentrations. CONCLUSION: Magainin on its own is highly embryotoxic. Its embryotoxicity is enhanced by cyclodextrin, albumin, H2O2 and acidification. Thus, magainin which has antibacterial, antifungal and antiprotozoal activity may also have a potential role as a contraceptive agent. The harmful effects of various concentrations of the exogenous H2O2 on 2-cell stage mouse embryos are reported here, to the best of our knowledge, for the first time.

Key words: cyclodextrin/embryotoxicity/hydrogen peroxide/magainine-2-amide/preimplantation mouse embryo

3 To whom correspondence should be addressed at: Center of Biostructure, Medical Academy, Chalubinski St. 5, 02–004 Warsaw, Poland. E-mail address: wsawicki{at}ib.amwaw.edu.pl


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