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      Medical-Grade Silicone Coated with Rhamnolipid R89 Is Effective against Staphylococcus spp. Biofilms

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          Abstract

          Staphylococcus aureus and Staphylococcus epidermidis are considered two of the most important pathogens, and their biofilms frequently cause device-associated infections. Microbial biosurfactants recently emerged as a new generation of anti-adhesive and anti-biofilm agents for coating implantable devices to preserve biocompatibility. In this study, R89 biosurfactant (R89BS) was evaluated as an anti-biofilm coating on medical-grade silicone. R89BS is composed of homologues of the mono- (75%) and di-rhamnolipid (25%) families, as evidenced by mass spectrometry analysis. The antimicrobial activity against Staphylococcus spp. planktonic and sessile cells was evaluated by microdilution and metabolic activity assays. R89BS inhibited S. aureus and S. epidermidis growth with minimal inhibitory concentrations (MIC 99) of 0.06 and 0.12 mg/mL, respectively and dispersed their pre-formed biofilms up to 93%. Silicone elastomeric discs (SEDs) coated by R89BS simple adsorption significantly counteracted Staphylococcus spp. biofilm formation, in terms of both built-up biomass (up to 60% inhibition at 72 h) and cell metabolic activity (up to 68% inhibition at 72 h). SEM analysis revealed significant inhibition of the amount of biofilm-covered surface. No cytotoxic effect on eukaryotic cells was detected at concentrations up to 0.2 mg/mL. R89BS-coated SEDs satisfy biocompatibility requirements for leaching products. Results indicate that rhamnolipid coatings are effective anti-biofilm treatments and represent a promising strategy for the prevention of infection associated with implantable devices.

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          Most cited references42

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          Microbial biosurfactants production, applications and future potential.

          Microorganisms synthesise a wide range of surface-active compounds (SAC), generally called biosurfactants. These compounds are mainly classified according to their molecular weight, physico-chemical properties and mode of action. The low-molecular-weight SACs or biosurfactants reduce the surface tension at the air/water interfaces and the interfacial tension at oil/water interfaces, whereas the high-molecular-weight SACs, also called bioemulsifiers, are more effective in stabilising oil-in-water emulsions. Biosurfactants are attracting much interest due to their potential advantages over their synthetic counterparts in many fields spanning environmental, food, biomedical, and other industrial applications. Their large-scale application and production, however, are currently limited by the high cost of production and by limited understanding of their interactions with cells and with the abiotic environment. In this paper, we review the current knowledge and the latest advances in biosurfactant applications and the biotechnological strategies being developed for improving production processes and future potential.
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            Rhamnolipids: diversity of structures, microbial origins and roles

            Rhamnolipids are glycolipidic biosurfactants produced by various bacterial species. They were initially found as exoproducts of the opportunistic pathogen Pseudomonas aeruginosa and described as a mixture of four congeners: α-L-rhamnopyranosyl-α-L-rhamnopyranosyl-β-hydroxydecanoyl-β-hydroxydecanoate (Rha-Rha-C10-C10), α-L-rhamnopyranosyl-α-L-rhamnopyranosyl-β-hydroxydecanoate (Rha-Rha-C10), as well as their mono-rhamnolipid congeners Rha-C10-C10 and Rha-C10. The development of more sensitive analytical techniques has lead to the further discovery of a wide diversity of rhamnolipid congeners and homologues (about 60) that are produced at different concentrations by various Pseudomonas species and by bacteria belonging to other families, classes, or even phyla. For example, various Burkholderia species have been shown to produce rhamnolipids that have longer alkyl chains than those produced by P. aeruginosa. In P. aeruginosa, three genes, carried on two distinct operons, code for the enzymes responsible for the final steps of rhamnolipid synthesis: one operon carries the rhlAB genes and the other rhlC. Genes highly similar to rhlA, rhlB, and rhlC have also been found in various Burkholderia species but grouped within one putative operon, and they have been shown to be required for rhamnolipid production as well. The exact physiological function of these secondary metabolites is still unclear. Most identified activities are derived from the surface activity, wetting ability, detergency, and other amphipathic-related properties of these molecules. Indeed, rhamnolipids promote the uptake and biodegradation of poorly soluble substrates, act as immune modulators and virulence factors, have antimicrobial activities, and are involved in surface motility and in bacterial biofilm development.
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              Healthcare-associated infections, medical devices and biofilms: risk, tolerance and control.

              Biofilms are of great importance in infection control and healthcare-associated infections owing to their inherent tolerance and 'resistance' to antimicrobial therapies. Biofilms have been shown to develop on medical device surfaces, and dispersal of single and clustered cells implies a significant risk of microbial dissemination within the host and increased risk of infection. Although routine microbiological testing assists with the diagnosis of a clinical infection, there is no 'gold standard' available to reveal the presence of microbial biofilm from samples collected within clinical settings. Furthermore, such limiting factors as viable but non-culturable micro-organisms and small-colony variants often prevent successful detection. In order to increase the chances of detection and provide a more accurate diagnosis, a combination of microbiological culture techniques and molecular methods should be employed. Measures such as antimicrobial coating and surface alterations of medical devices provide promising opportunities in the prevention of biofilm formation on medical devices.
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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                Molecules
                Molecules
                molecules
                Molecules
                MDPI
                1420-3049
                25 October 2019
                November 2019
                : 24
                : 21
                : 3843
                Affiliations
                [1 ]Department of Pharmaceutical Sciences, Università del Piemonte Orientale “A. Avogadro”, 28100 Novara, Italy; chiara.ceresa@ 123456uniupo.it (C.C.); erica.tambone@ 123456unitn.it (E.T.); emanuele.fedeli@ 123456uniupo.it (E.F.); gianna.allegrone@ 123456uniupo.it (G.A.); maurizio.rinaldi@ 123456uniupo.it (M.R.)
                [2 ]BIOtech Center for Biomedical Technologies, Department of Industrial Engineering, Università di Trento, 38123 Trento, Italy; francesco.tessarolo@ 123456unitn.it (F.T.); devid.maniglio@ 123456unitn.it (D.M.); giandomenico.nollo@ 123456unitn.it (G.N.)
                [3 ]Healthcare Research and Innovation Program (IRCS-FBK-PAT), Bruno Kessler Foundation, 38123 Trento, Italy
                [4 ]Department of Mechanical and Aerospace Engineering, Politecnico di Torino, 10129 Turin, Italy; irene.carmagnola@ 123456polito.it (I.C.); valeria.chiono@ 123456polito.it (V.C.)
                [5 ]Section of Electron Microscopy, Department of Medicine Laboratory, Azienda Provinciale per i Servizi Sanitari di Trento, 38123 Trento, Italy; iole.caola@ 123456gmail.com
                Author notes
                [* ]Correspondence: letizia.fracchia@ 123456uniupo.it ; Tel.: +39-0321-375839
                [†]

                These Authors equally contributed to this work.

                Author information
                https://orcid.org/0000-0003-4022-5602
                https://orcid.org/0000-0002-1653-861X
                https://orcid.org/0000-0001-6080-317X
                https://orcid.org/0000-0003-2067-7732
                https://orcid.org/0000-0003-1346-1634
                https://orcid.org/0000-0003-3505-1919
                Article
                molecules-24-03843
                10.3390/molecules24213843
                6864460
                31731408
                857387c5-a22a-4389-afaa-bfc922e9fa65
                © 2019 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 12 September 2019
                : 22 October 2019
                Categories
                Article

                biofilm,biosurfactants,staphylococcus spp.,antimicrobial,anti-adhesive,anti-biofilm,biomaterials,silicone,medical devices

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