Carbohydrate Polymers 127 (2015) 451–461 Contents lists available at ScienceDirect Carbohydrate Polymers j ourna l ho me page: www.elsevier.com/locate/carbpol Incorporation of antimicrobial peptides on functionalized cotton gauzes for medical applications A.P. Gomes a , J.F. Mano b,c , J.A. Queiroz d , I.C. Gouveia a,∗ a FibEnTech – Fiber Materials and Environmental Technologies – Research Unit, Faculty of Engineering, University of Beira Interior, 6201-001 Covilhã, Portugal b 3B’s Research Group – Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, 4806-909 Taipas, Guimarães, Portugal c ICVS/3B’s – PT Government Associate Laboratory, Braga/Guimarães, Portugal d Health Sciences Research Centre, University of Beira Interior, 6201-001 Covilhã, Portugal a r t i c l e i n f o Article history: Received 29 September 2014 Received in revised form 25 March 2015 Accepted 31 March 2015 Available online 14 April 2015 Keywords: Antimicrobial peptides hBD-1 -Defensin-1 Human Dermaseptin Cys-LC-LL-37 Magainin 1 Biocompatibility Wound-dressing a b s t r a c t A large group of low molecular weight natural compounds that exhibit antimicrobial activity has been isolated from animals and plants during the past two decades. Among them, peptides are the most widespread resulting in a new generation of antimicrobial agents with higher specific activity. In the present study we have developed a new strategy to obtain antimicrobial wound-dressings based on the incorporation of antimicrobial peptides into polyelectrolyte multilayer films built by the alternate deposition of polycation (chitosan) and polyanion (alginic acid sodium salt) over cotton gauzes. Energy dispersive X ray microanalysis technique was used to determine if antimicrobial peptides penetrated within the films. FTIR analysis was performed to assess the chemical linkages, and antimicrobial assays were performed with two strains: Staphylococcus aureus (Gram-positive bacterium) and Klebsiella pneu- monia (Gram-negative bacterium). Results showed that all antimicrobial peptides used in this work have provided a higher antimicrobial effect (in the range of 4 log–6 log reduction) for both microor- ganisms, in comparison with the controls, and are non-cytotoxic to normal human dermal fibroblasts at the concentrations tested. © 2015 Elsevier Ltd. All rights reserved. 1. Introduction Several authors found that there was a significant absorption of antibiotic, when it is placed directly on the wound as a cream, which may increase the risk of cytotoxicity of the treated tissues, because in this case easily excessive amounts that can be used and it is difficult to control the optimal amount of cream. (Boosalis, McCall, Ahrenholz, Solem, & McClain, 1987; Mi et al., 2002; Wang, Wang, Zhang, Zapatasirvent, & Davies, 1985). Likewise, it is important to develop a method to control the release of antimicrobial agents. It also has been reported that higher concentrations of some compounds are toxic to tissue and may be a burden to organs or lead to the development of antibiotic resistance (Boateng, Matthews, Stevens, & Eccleston, 2008; Dave, Joshi, & Venugopalan, 2012; Hidalgo & Dominguez, 1998). Compounds most commonly incorporated into dressings to control or prevent infection are silver (Boateng et al., 2008), povidone–iodine (Misra & Nanchahal, ∗ Corresponding author. Tel.: +35 1917248532. E-mail address: igouveia@ubi.pt (I.C. Gouveia). 2003) and polyhexamethylene biguanide (Motta, Milne, & Corbett, 2004). On the other hand, semi-solid preparations such as silver sulphadiazine cream (Hudspith & Rayatt, 2004) and silver nitrate ointment (Moir & Serra, 2012) are used to treat bacterial infection on the surface of the wound but direct application onto open wounds can be very painful (Thakoersing et al., 2012) and the scientific evidence for the efficacy of these agents in wounds is scarce. Common topical antibiotics also include mupirocin (Rode, Hanslo, Dewet, Millar, & Cywes, 1989), neosporin (Sinha, Agarwal, & Agarwal, 1997) and tetracycline (Kumar, Bai, & Krishnan, 2004). However, these antibiotics are ineffective when resistant bacteria colonize the wound (Cookson, 1998; Hetem & Bonten, 2013). Moreover, it is important that slow release of antimicrobial agent from wound dressing have the advantage of treating infected wounds in a mild way (Elsner, Berdicevsky, & Zilberman, 2011; Kostenko, Lyczak, Turner, & Martinuzzi, 2010). Since the beginning of the antibiotic era in the 1940s, the use of antibiotics has resulted in the continual emergence of resistant strains of bacteria, further complicating the clearance of infection in cutaneous wounds (Gibson et al., 2012). Therefore, a new and innovative strategy is needed to combat infected cutaneous http://dx.doi.org/10.1016/j.carbpol.2015.03.089 0144-8617/© 2015 Elsevier Ltd. All rights reserved.