UNCORRECTED PROOF Nanomedicine: Nanotechnology, Biology, and Medicine xxx (2017) xxx-xxx Contents lists available at ScienceDirect Nanomedicine: Nanotechnology, Biology, and Medicine journal homepage: www.elsevier.com Immobilization of bacteriophage in wound-dressing nanostructure Frederico Nogueira a, b , Natia Karumidze c , Ia Kusradze c , Marina Goderdzishvili c , Pilar Teixeira d , Isabel C. Gouveia b, ⁎ a CICS-UBI – Health Sciences Research Centre, University of Beira Interior, Portugal b FibEnTech – Fiber Materials and Environmental Technologies, University of Beira Interior, Portugal c G. Eliava Institute of Bacteriophages, Microbiology and Virology, Tbilisi, Georgia d Institute for Biotechnology and Bioengineering (IBB), Portugal ARTICLE INFO Article history: Received 25 March 2017 Accepted 9 August 2017 Available online xxx Key words: Bacteriophages Antimicrobial agents Surface immobilization Electrospinning Pseudomonas aeruginosa ABSTRACT Opportunistic bacteria that cause life-threatening infections are still a central problem associated with a healthcare setting. Bacteriophage capsid immobilization on nanostructured polymers maximizes its tail expo- sure and looks promising in applications toward skin-infections as alternative to antibiotics standardly used. The main goal of this work was to investigate the covalent immobilization of vB_Pae_Kakheti25 bacterio- phage capsid on polycaprolactone (PCL) nanofibers (non-woven textile), as a potential effective antimicro- bial, laundry resistant and non-toxic dressing for biomedical use. Surface analyses showed that the immobi- lization of vB_Pae_Kakheti25 bacteriophage capsid on PCL nanofibres oriented bacteriophage tails to inter- act with bacteria. Furthermore, antimicrobial assays showed a very effective 6 log bacterial reduction, which was equivalent to 99.9999%, after immediate and 2 hours of contact, even following 25 washing cycles (due to covalent bond). The activity of PCL-vB_Pae_Kakheti25 against P. aeruginosa was immediate and its re- duction was complete. © 2017. The skin of patients with inflammatory skin-diseases alongside with chronic or burn wounds and exit-sites of catheters is partic- ularly susceptible to infection by different microorganisms. Oppor- tunistic pathogens are the cause of skin diseases, infections, and the inability of chronic wounds to heal. 1–4 They are capable of produc- ing virulence factors, including enzymes that promote tissue invasion and extracellular polymers, which form the biofilm that contributes to the perpetuation of skin inflammation, even in normal-appearing skin. Fortunately, the majority of our resident skin microorganisms are non-pathogenic and many of these contribute to maintaining health. 1 Accordingly, skin-disease/injury management demands an integrated approach aimed not only at diminishing infection but also at regulat- ing the skin microbiome. 2,5 P. aeruginosa is the most common infectious agent among Pseudomonas spp. As a versatile and opportunistic microorganism it can colonize the skin, soft tissue, gastrointestinal tract, armpits, eye and ear. 6–8 P. aeruginosa is the agent responsible for the most com- mon infections under hospital settings, through catheter and ventila- tor contaminations leading to nosocomial infections, such as pneumo- nia, urinary tract and wound burn infections, as well as bacteremia, especially in patients with diabetes or immunodeficiency. 8 The major Acknowledgments: The authors acknowledge the Fundação para a Ciência e Tecnologia (FCT) for the PhD grant SFRH/BD/91444/2012 and Programa Operacional Capital Humano (POCH) and European Union for co-funding the work. ⁎ Corresponding author at: FibEnTech R&D Fiber Materials and Environmental Technologies, University of Beira Interior, 6201-001 Covilhã, Portugal. Email address: igouveia@ubi.pt (I.C. Gouveia) concerns about the control of nosocomial infections vary from the problems of drug safety associated with a high human toxicity, the long-term and large scale application of broad-spectrum antibiotic drugs, to the increased resistance to conventional therapies. These in- fections tend to chronicity and may fail to be treated with almost any combination of antibiotics, showing mortalities up to 61%. 9 The com- bination therapy to fight P. aeruginosa infections is very difficult to achieve, due to the compromised immune system of the majority of in- fected patients, and the intrinsic resistance of microorganisms to vari- ous antibiotics. 10 Recent research has been conducted on the three groups of natu- rally occurring antimicrobials as novel alternatives to antibiotics: bac- teriophages, bacterial cell wall hydrolases (BCWHs), and antimicro- bial peptides (AMPs). 11 Among them, bacteriophages are the most highly specific toward both Gram-positive and Gram-negative bac- teria and they are also highly efficient and relatively cost-effective. In contrast, AMPs have a broad-spectrum against bacteria and fun- gus, low level of induced resistance, but may cause toxicity at high doses in order to be efficient, and are more costly to produce. 11 BCWH has limitations toward Gram-negative bacteria, as a result of the pres- ence of the outer membrane, and important Gram-positive pathogens like S. aureus are already resistant to lysozymes. To overcome the changing tide of nosocomial diseases and increasing reports of mi- croorganism-acquired resistances, recently the United States, Canada and European countries have started to take a close interest in bac- teriophage-based therapies, 12,13 in which they act without mecha- nism-based host toxicity. The bacteriophage vB_Pae_Kakheti25 has a potent activity against P. aeruginosa and appears as an alternative approach to con https://doi.org/10.1016/j.nano.2017.08.008 1549-9634/© 2017.