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Novel gramicidin formulations in cationic lipid as broad-spectrum microbicidal agents

Authors Ragioto D, Carrasco L, Carmona-Ribeiro AM

Received 1 April 2014

Accepted for publication 3 May 2014

Published 30 June 2014 Volume 2014:9(1) Pages 3183—3192


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Danielle AMT Ragioto, Letícia DM Carrasco, Ana M Carmona-Ribeiro

Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, Brazil

Abstract: Dioctadecyldimethylammonium bromide (DODAB) is an antimicrobial lipid that can be dispersed as large closed bilayers (LV) or bilayer disks (BF). Gramicidin (Gr) is an antimicrobial peptide assembling as channels in membranes and increasing their permeability towards cations. In mammalian cells, DODAB and Gr have the drawbacks of Gram-positive resistance and high toxicity, respectively. In this study, DODAB bilayers incorporating Gr showed good antimicrobial activity and low toxicity. Techniques employed were spectroscopy, photon correlation spectroscopy for sizing and evaluation of the surface potential at the shear plane, turbidimetric detection of dissipation of osmotic gradients in LV/Gr, determination of bacterial cell lysis, and counting of colony-forming units. There was quantitative incorporation of Gr and development of functional channels in LV. Gr increased the bilayer charge density in LV but did not affect the BF charge density, with localization of Gr at the BF borders. DODAB/Gr formulations substantially reduce Gr toxicity against eukaryotic cells and advantageously broaden the antimicrobial activity spectrum, effectively killing Escherichia coli and Staphylococcus aureus bacteria with occurrence of cell lysis.

Keywords: antimicrobial peptides, cationic bilayers, bilayer fragments, dioctadecyldimethylammonium bromide, gramicidin A, self-assembly

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