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Layer-by-layer assembling of hyaluronan/chitosan nanofilms and their antibacterial effect: a study using Staphylococcus aureus and Pseudomonas aeruginosa

2016; Frontiers Media; Volume: 4; Linguagem: Inglês

10.3389/conf.fbioe.2016.01.02566

2296-4185

Jacobo Hernández‐Montelongo, Lucchesi Eliane, Gonzalez-Bravo Ismael, Macedo Waldemar, Prasana Sahoo, Nascimento Vicente, Beppu Marisa, Moraes Angela, Cotta Monica,

Advanced Drug Delivery Systems

Event Abstract Back to Event Layer-by-layer assembling of hyaluronan/chitosan nanofilms and their antibacterial effect: a study using Staphylococcus aureus and Pseudomonas aeruginosa Jacobo Hernandez-Montelongo1, Eliane Lucchesi2, Ismael Gonzalez-Bravo3, Waldemar Macedo3, Prasana Sahoo1, Vicente Nascimento4, Marisa Beppu4, Angela Moraes4 and Monica Cotta1 1 Instituto de Física Gleb Wataghin, Department of Applied Physics, Brazil 2 Itibanyl Produtos Especiais LTDA, Brazil 3 Centro de Desenvolvimento da Tecnologia Nuclear, Department of Nanotechnology, Brazil 4 Faculdade de Engenharia Química, Department of Materials Engineering and Bioprocess, Brazil The layer-by-layer (LbL) is a simple, versatile and cost-effective technique to form thin films on different materials. This method consists of alternating physisorption of oppositely charged polyelectrolytes[1]. As hyaluronan (HA) and chitosan (CHI) are oppositely charged biopolymers (negative and positive, respectively), they are excellent candidates to be used for surface coatings obtained by this technique (Fig. 1A)[2],[3]. Due to the antimicrobial characteristics of CHI by NH3+ groups, bilayers of HA/CHI can be used as antibacterial coatings[4]. In that sense, this work is about the synthesis of antibacterial nanofilms of HA/CHI biopolymers assembled by LbL, varying specific pH values of polyelectrolyte solutions (Fig. 1B). HA/CHI assemblies can be monitored by measuring their respective surface properties, in each alternating deposition. The final surface physicochemical properties can be correlated with their obtained antibacterial properties. In this work, we considered two human pathogenic microorganisms, Staphylococcus aureus and Pseudomonas aeruginosa. The antibacterial effect of HA/CHI nanofilms was evaluated using the spread plate counting method. These results were correlated with the morphology of nanofilms (characterized using SEM and AFM), as well as with their chemical properties studied by UV-Vis and XPS spectroscopy. Fig. 1C shows the antibacterial effect of the HA/CHI nanofilms. In the case of S. aureus assay, the strain was very sensible to the pH of synthesis of samples, which is strongly related with the free ammonium groups on the top CHI layer of samples. Compared to the control, the Si substrate, sample pH = 4.5 reduced 1 and 2 orders of magnitude of CFU/cm2 at 4 and 8 h of cell culture, respectively. The antibacterial effect of the pH = 3 was very notorious in both times of culture, reducing 3 orders of magnitude for 4 h and 5 orders for 8 h. In the case of pH = 2, there was not important reduction of the bacterial density, for both times culture. In the case of P. aeruginosa assay, three samples did not show important antibacterial effect. The resistance of P. aeruginosa can be explained by its outer membrane, which would protect its cell wall from the killer contact to of free protonated ammonium groups of films. Figure 1. A) Schematic representation of the buildup of HA/CHI multilayer thin films by LbL: immersion of the charged substrate in the correspondent polysaccharide electrolyte, alternating with rinsing steps. B) Ionization curves of HA and CHI obtained by the Henderson-Hasselbach equation; pKa of HA is 3.0 while that of CHI is 6.5. The pH values of nanofilms synthesis condition are also indicated: 4.5, 3 and 2. C) Antibacterial effect of HA/CHI nanofilms. The number of viable bacteria on the samples at 4 and 8 hours as determined using the spread plate method. ** denotes a significant difference compared to the Si substrate at 4h (p < 0.01); ## denotes a significant difference compared to the Si substrate at 8h (p < 0.01). Analytical Resources and Calibration Laboratory from UNICAMP; Multi-user Lab from UNICAMP; National Nanotechnology Laboratory; São Paulo Research Foundation (FAPESP); National Council for Scientific and Technological Development (CNPq); Coordenadoria de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)References:[1] J. Borges, and J. F. Mano, “Molecular Interactions Driving the Layer-by-Layer Assembly of Multilayers”, Chem. Rev. 114 (2014) 8883-8942[2] S. N. Chirkov, “The Antiviral Activity of Chitosan (Review)” Appl. Biochem. Microbiol. 38 (2002) 1-8.[3] J. Necas, L. Bartosikova, P. Brauner, and J. Kolar, “Hyaluronic acid (hyaluronan): a review”, J. Vet. Med. 35 (2008) 397-411[4] J. Hernandez-Montelongo, V. Nascimento, D. Murillo, T. Taketa, P. Sahoo, A. de Souza, M. Beppu, M. Cotta. C, “Nanofilms of hyaluronan/chitosan assembled layer-by-layer: An antibacterial surface for Xylella fastidiosa”, Carbohyd. Polym. 136 (2016) 1-11 Keywords: Bacteria, in vitro, biomaterial, Ultra-thin coating Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016. Presentation Type: Poster Topic: Anti-infective biomaterials Citation: Hernandez-Montelongo J, Lucchesi E, Gonzalez-Bravo I, Macedo W, Sahoo P, Nascimento V, Beppu M, Moraes A and Cotta M (2016). Layer-by-layer assembling of hyaluronan/chitosan nanofilms and their antibacterial effect: a study using Staphylococcus aureus and Pseudomonas aeruginosa. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.02566 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 27 Mar 2016; Published Online: 30 Mar 2016. Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Jacobo Hernandez-Montelongo Eliane Lucchesi Ismael Gonzalez-Bravo Waldemar Macedo Prasana Sahoo Vicente Nascimento Marisa Beppu Angela Moraes Monica Cotta Google Jacobo Hernandez-Montelongo Eliane Lucchesi Ismael Gonzalez-Bravo Waldemar Macedo Prasana Sahoo Vicente Nascimento Marisa Beppu Angela Moraes Monica Cotta Google Scholar Jacobo Hernandez-Montelongo Eliane Lucchesi Ismael Gonzalez-Bravo Waldemar Macedo Prasana Sahoo Vicente Nascimento Marisa Beppu Angela Moraes Monica Cotta PubMed Jacobo Hernandez-Montelongo Eliane Lucchesi Ismael Gonzalez-Bravo Waldemar Macedo Prasana Sahoo Vicente Nascimento Marisa Beppu Angela Moraes Monica Cotta Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.