Hyaluronan/chitosan nanofilms assembled layer-by-layer and their antibacterial effect: A study using Staphylococcus aureus and Pseudomonas aeruginosa.

In the last few years, chitosan-based coatings have been proposed as antibacterial surfaces for biomedical devices in order to prevent nosocomial infections. In that sense, this work reports the optimized synthesis of hyaluronan/chitosan (HA/CHI) nanofilms assembled layer-by-layer in order to maximize the antibacterial effect for two important human pathogenic bacteria, Staphylococcus aureus and Pseudomonas aeruginosa. In this assembly, HA forms a soft, highly hydrated, and nontoxic film, whereas CHI shows the antimicrobial characteristics. Our HA/CHI nanofilm synthesis optimization was based on changing pH values of the biopolymer stem-solutions and the consequent variation of their ionization degree. Furthermore, the surface density of primary amino groups, which are related to the antibacterial effect, was also enhanced by increasing the number of HA/CHI bilayers. The antibacterial effect of HA/CHI nanofilms was evaluated by the spread plate counting method for both bacteria. 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, Kelvin Probe Force microscopy and XPS spectroscopy.

[1]  J. García-Ruíz,et al.  Chemical stabilization of porous silicon for enhanced biofunctionalization with immunoglobulin , 2012, Science and technology of advanced materials.

[2]  Joseph J. Richardson,et al.  Technology-driven layer-by-layer assembly of nanofilms , 2015, Science.

[3]  Y. Yun,et al.  The role of biomass in polyethylenimine-coated chitosan/bacterial biomass composite biosorbent fiber for removal of Ru from acetic acid waste solution. , 2014, Bioresource technology.

[4]  Duber M. Murillo,et al.  Nanofilms of hyaluronan/chitosan assembled layer-by-layer: An antibacterial surface for Xylella fastidiosa. , 2016, Carbohydrate polymers.

[5]  M. Kipper,et al.  Layer-by-layer assembly of polysaccharide-based polyelectrolyte multilayers: a spectroscopic study of hydrophilicity, composition, and ion pairing. , 2011, Biomacromolecules.

[6]  T. Suni,et al.  Effects of Plasma Activation on Hydrophilic Bonding of Si and SiO2 , 2002 .

[7]  M. Goosen,et al.  Antibacterial activity of chitin, chitosan and its oligomers prepared from shrimp shell waste , 2012 .

[8]  Wei He,et al.  Nanoscale neuro-integrative coatings for neural implants. , 2005, Biomaterials.

[9]  R. Auzély-Velty,et al.  Contact‐Killing Polyelectrolyte Microcapsules Based on Chitosan Derivatives , 2010 .

[10]  Jia-cong Shen,et al.  Construction of anti-adhesive and antibacterial multilayer films via layer-by-layer assembly of heparin and chitosan. , 2005, Biomaterials.

[11]  Peter Fredericks,et al.  Interactions between alginate and chitosan biopolymers characterized using FTIR and XPS. , 2007, Biomacromolecules.

[12]  I. Petrinic,et al.  Stability of a chitosan layer deposited onto a polyethylene surface , 2013 .

[13]  K. Neoh,et al.  Surface functionalization of titanium with hyaluronic acid/chitosan polyelectrolyte multilayers and RGD for promoting osteoblast functions and inhibiting bacterial adhesion. , 2008, Biomaterials.

[14]  T. D. Martins,et al.  Surface modification of polyelectrolyte multilayers by high radio frequency air plasma treatment , 2015 .

[15]  C. Jérôme,et al.  Chitosan-based biomaterials for tissue engineering , 2013 .

[16]  B. Iglewski,et al.  Quorum-Sensing Genes in Pseudomonas aeruginosa Biofilms: Their Role and Expression Patterns , 2001, Applied and Environmental Microbiology.

[17]  D. Paterson,et al.  Reducing the development of antibiotic resistance in critical care units. , 2011, Current Pharmaceutical Biotechnology.

[18]  S. Kannan,et al.  Update on bacterial nosocomial infections. , 2012, European review for medical and pharmacological sciences.

[19]  G. Prestwich,et al.  Layer by layer buildup of polysaccharide films: physical chemistry and cellular adhesion aspects. , 2004, Langmuir : the ACS journal of surfaces and colloids.

[20]  A. Packman,et al.  Pseudomonas aeruginosa Promotes Escherichia coli Biofilm Formation in Nutrient-Limited Medium , 2014, PloS one.

[21]  R. Shekhar,et al.  Chitosan nanocoating on cotton textile substrate using layer-by-layer self-assembly technique , 2011 .

[22]  C. Vasile,et al.  Chitosan/hyaluronic acid polyelectrolyte complex hydrogels in the management of burn wounds. , 2013, Revista medico-chirurgicala a Societatii de Medici si Naturalisti din Iasi.

[23]  Jian Ji,et al.  Construction of antibacterial multilayer films containing nanosilver via layer-by-layer assembly of heparin and chitosan-silver ions complex. , 2006, Journal of biomedical materials research. Part A.

[24]  Bin Li,et al.  Chitosan/phosvitin antibacterial films fabricated via layer-by-layer deposition. , 2014, International journal of biological macromolecules.

[25]  Timothy J. Foster,et al.  Adhesion, invasion and evasion: the many functions of the surface proteins of Staphylococcus aureus , 2013, Nature Reviews Microbiology.

[26]  F. Rossi,et al.  Porous silicon-cyclodextrin based polymer composites for drug delivery applications. , 2014, Carbohydrate polymers.

[27]  M. Rubner,et al.  Bioactive polyelectrolyte multilayers: hyaluronic acid mediated B lymphocyte adhesion. , 2010, Biomacromolecules.

[28]  Jean-Claude Voegel,et al.  Self‐Defensive Biomaterial Coating Against Bacteria and Yeasts: Polysaccharide Multilayer Film with Embedded Antimicrobial Peptide , 2013 .

[29]  K. Kerr,et al.  Association between healthcare water systems and Pseudomonas aeruginosa infections: a rapid systematic review. , 2014, The Journal of hospital infection.

[30]  Michael F. Rubner,et al.  Controlling Bilayer Composition and Surface Wettability of Sequentially Adsorbed Multilayers of Weak Polyelectrolytes , 1998 .