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Encapsulation in Polymeric Microparticles Improves Daptomycin Activity Against Mature Staphylococci Biofilms—a Thermal and Imaging Study

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Abstract

Eradication of Gram-positive biofilms is a critical aspect in implant-associated infection treatment. Although antibiotic-containing particulate carriers may be a promising strategy for overcoming biofilm tolerance, the assessment of their interaction with biofilms has not been fully explored. In the present work, the antibiofilm activity of daptomycin- and vancomycin-loaded poly(methyl methacrylate) (PMMA) and PMMA-Eudragit RL 100 (EUD) microparticles against methicillin-resistant Staphylococcus aureus (MRSA) and polysaccharide intercellular adhesin-positive S. epidermidis biofilms was investigated using isothermal microcalorimetry (IMC) and fluorescence in situ hybridization (FISH). The minimal biofilm inhibitory concentrations (MBIC) of MRSA biofilms, as determined by IMC, were 5 and 20 mg/mL for daptomycin- and vancomycin-loaded PMMA microparticles, respectively. S. epidermidis biofilms were less susceptible, with a MBIC of 20 mg/mL for daptomycin-loaded PMMA microparticles. Vancomycin-loaded microparticles were ineffective. Adding EUD to the formulation caused a 4- and 16-fold reduction of the MBIC values of daptomycin-loaded microparticles for S. aureus and S. epidermidis, respectively. FISH corroborated the IMC results and provided additional insights on the antibiofilm effect of these particles. According to microscopic analysis, only daptomycin-loaded PMMA-EUD microparticles were causing a pronounced reduction in biofilm mass for both strains. Taken together, although IMC indicated that a biofilm inhibition was achieved, microscopy showed that the biofilm was not eradicated and still contained FISH-positive, presumably viable bacteria, thus indicating that combining the two techniques is essential to fully assess the effect of microparticles on staphylococcal biofilms.

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Acknowledgements

The authors acknowledge Novartis Pharma (Basel, Switzerland) for the gift of daptomycin. The paper is based upon work from COST action (TD1305, IPROMEDAI).

Funding

This work was supported by the Portuguese government (Fundação para a Ciência e a Tecnologia) and FEDER Grants SFRH/BD/69260/2010, research projects proj. 6818 FCT/DAAD and EXCL/CTM-NAN/0166/2012 and strategic project iMed.ULisboa (UID/DTP/04138/2013).

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Author notes

  1. Inês Santos Ferreira and Judith Kikhney contributed equally to this work.

Authors and Affiliations

  1. Research Institute for Medicines (iMed.ULisboa), Faculdade de Farmácia, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003, Lisbon, Portugal

    Inês Santos Ferreira, Lídia M. D. Gonçalves, Ana Francisca Bettencourt & António J. Almeida

  2. Biofilmcenter, Deutsches Herzzentrum Berlin, Charité—Universitätsmedizin Berlin, Hindenburgdamm 30, 12203, Berlin, Germany

    Judith Kikhney, Laura Kursawe, Stefanie Kasper & Annette Moter

  3. Institute for Microbiology and Hygiene, Charité—Universitätsmedizin Berlin, Hindenburgdamm 30, 12203, Berlin, Germany

    Judith Kikhney

  4. Center for Musculoskeletal Surgery, Charité—Universitätsmedizin Berlin, Free and Humboldt-University of Berlin, Charitéplatz 1, 10117, Berlin, Germany

    Andrej Trampuz

Authors
  1. Inês Santos Ferreira
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  2. Judith Kikhney
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Corresponding authors

Correspondence to Ana Francisca Bettencourt or António J. Almeida.

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Santos Ferreira, I., Kikhney, J., Kursawe, L. et al. Encapsulation in Polymeric Microparticles Improves Daptomycin Activity Against Mature Staphylococci Biofilms—a Thermal and Imaging Study. AAPS PharmSciTech 19, 1625–1636 (2018). https://doi.org/10.1208/s12249-018-0974-7

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