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Antibiotics
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Bacterial Extracellular Vesicles: Vehicles for Pathogenesis and Antibiotic Resistance
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Bacterial Extracellular Vesicles: Vehicles for Pathogenesis and Antibiotic Resistance

A special issue of Antibiotics (ISSN 2079-6382). This special issue belongs to the section "Mechanism and Evolution of Antibiotic Resistance".

Deadline for manuscript submissions: 30 June 2026 | Viewed by 4745

Special Issue Editor

Prof. Dr. Eva Sapi

E-Mail Website
Guest Editor
Department of Biology and Environment Science, University of New Haven, West Haven, CT 06516, USA
Interests: Lyme disease; Borrelia burgdorferi; host pathogen interaction; biofilm; antibiotic resistance; outer membrane vesicles; infection origins of breast cancer
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Recent research has revealed that bacterial extracellular vesicles (BEVs) play a central role in intercellular communication by transporting proteins, lipids, nucleic acids, and other bioactive molecules. These vesicles are now recognized as important mediators in a range of physiological and pathological processes, including immune modulation, inflammation, cancer progression, tissue repair, and angiogenesis.

Importantly, BEVs have also been implicated in the development and dissemination of antibiotic resistance. They can carry resistance genes and enzymes such as β-lactamases, facilitate horizontal gene transfer, and influence bacterial adaptation under antibiotic stress. This highlights their potential not only as key players in resistance mechanisms but also as possible targets for novel therapeutic interventions.

This Special Issue will provide a comprehensive overview of our current understanding of bacterial extracellular vesicles, with a focus on their roles in pathogenesis and antimicrobial resistance. We welcome contributions that address, but are not limited to, the following topics:

  • Mechanisms behind BEV formation, release, and regulation;
  • BEV involvement in host–pathogen interactions;
  • Roles of BEVs in horizontal gene transfer and resistance gene dissemination;
  • BEVs as carriers of antibiotic resistance determinants;
  • The impact of BEVs on microbial survival and adaptation under antibiotic pressure;
  • The diagnostic and therapeutic potential of BEVs in infectious diseases;
  • Novel approaches to targeting or engineering BEVs for antimicrobial applications;
  • Methodologies for isolating, characterizing, and analyzing BEVs;
  • Comprehensive reviews on BEV biology and clinical relevance.

For this Special Issue, we welcome manuscript submissions of various types, such as original research articles, short communications, comprehensive reviews, case reports, and perspectives.

Prof. Dr. Eva Sapi
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 250 words) can be sent to the Editorial Office for assessment.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Antibiotics is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2900 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • bacterial extracellular vesicles (BEVs)
  • outer membrane vesicles (OMVs)
  • antibiotic resistance
  • pathogenesis
  • host-pathogen interaction

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Published Papers (3 papers)

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Research

14 pages, 2357 KB  
Article
Oxidative Stress Reshapes Porphyromonas gingivalis Outer Membrane Vesicles and Impairs OMV-Mediated Invasion and Persistence in Trophoblast Cells
by Ailén Fretes, Brenda Lara, Mateo N. Diaz Appella, Carolina López, Claudia Pérez Leirós, Paula M. Tribelli and Vanesa Hauk
Antibiotics 2026, 15(2), 152; https://doi.org/10.3390/antibiotics15020152 - 2 Feb 2026
Cited by 1 | Viewed by 982
Abstract
Background: Porphyromonas gingivalis outer membrane vesicles (OMVs) are key mediators of host–pathogen interactions and have been implicated in both periodontal disease and systemic conditions, including pregnancy complications. Although OMV production and cargo are known to be influenced by environmental stress, how oxidative [...] Read more.
Background: Porphyromonas gingivalis outer membrane vesicles (OMVs) are key mediators of host–pathogen interactions and have been implicated in both periodontal disease and systemic conditions, including pregnancy complications. Although OMV production and cargo are known to be influenced by environmental stress, how oxidative stress reshapes P. gingivalis OMVs and their functional impact on trophoblast cells remains poorly understood. Here, we investigated how exposure to hydrogen peroxide (H2O2) affects OMV biogenesis, composition, and their ability to modulate bacterial invasion in trophoblast cells. Methods: P. gingivalis was cultured anaerobically and exposed to 30 mM H2O2 during the final 24 h of growth. OMVs were isolated by differential ultracentrifugation and characterized by nanoparticle tracking analysis and transmission electron microscopy and OMV protein cargo was analyzed by proteomics. Functional effects were assessed using invasion and persistence assays in HTR-8/SVneo trophoblast cells pretreated with OMVs. Results: Oxidative stress did not significantly alter total OMV yield but resulted in smaller vesicles (control OMV 168.2 ± 8.7 nm vs. OMV from H2O2-treated cultures 130.0 ± 13.8 nm) with reduced negative surface charge and increased membrane-associated FM4-64 fluorescence. Proteomic analysis revealed a remodeling of the OMV protein cargo under oxidative stress, including the selective enrichment of a von Willebrand factor type A domain-containing protein. Functionally, OMVs from control cultures led to a 2.5-fold increase in P. gingivalis invasion and a 4-fold increase in intracellular persistence in trophoblast cells, whereas OMVs produced under oxidative stress failed to promote these processes. Conclusions: Together, these findings highlight oxidative stress as a key determinant of OMV-mediated host–pathogen interactions at the maternal–fetal interface. Full article
(This article belongs to the Special Issue Bacterial Extracellular Vesicles: Vehicles for Pathogenesis and Antibiotic Resistance)
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15 pages, 1518 KB  
Article
Biophysical Features of Outer Membrane Vesicles (OMVs) from Pathogenic Escherichia coli: Methodological Implications for Reproducible OMV Characterization
by Giorgia Barbieri, Linda Maurizi, Maurizio Zini, Federica Fratini, Agostina Pietrantoni, Ilaria Bellini, Serena Cavallero, Eleonora D’Intino, Federica Rinaldi, Paola Chiani, Valeria Michelacci, Stefano Morabito, Barbara Chirullo and Catia Longhi
Antibiotics 2026, 15(2), 117; https://doi.org/10.3390/antibiotics15020117 - 26 Jan 2026
Cited by 2 | Viewed by 1311
Abstract
Background/Objectives: Bacterial outer membrane vesicles (OMVs) play a role in bacterial communication, virulence, antimicrobial resistance, and host–pathogen interaction. OMV isolation is a key step for studying these particles’ functions; nevertheless, isolation procedures can greatly influence the yield, purity, and structural integrity of [...] Read more.
Background/Objectives: Bacterial outer membrane vesicles (OMVs) play a role in bacterial communication, virulence, antimicrobial resistance, and host–pathogen interaction. OMV isolation is a key step for studying these particles’ functions; nevertheless, isolation procedures can greatly influence the yield, purity, and structural integrity of OMVs, thereby affecting downstream biological analyses and functional interpretation. Methods: In this study, we compared the efficacy of two OMV isolation techniques, differential ultracentrifugation (dUC) and size-exclusion chromatography (SEC), in separating and concentrating vesicles produced by two Escherichia coli strains belonging to uropathogenic (UPEC) and Shiga toxin-producing (STEC) pathotypes. The isolated OMVs were characterized using a multi-analytical approach including transmission and scanning electron microscopy (TEM, SEM), nanoparticle tracking analysis (NTA), dynamic light scattering (DLS), ζ-potential measurement, and protein quantification to assess the purity of the preparations. Results: Samples obtained by dUC exhibited higher total protein content, broader particle size distributions, and more pronounced contamination by non-vesicular material. In contrast, SEC yielded morphologically homogeneous and structurally well-preserved vesicles, higher particle-to-protein ratios, and lower total protein content, reflecting reduced co-isolation of protein aggregates. NTA and DLS analyses revealed polydisperse populations in samples obtained with both isolation methods, with DLS measurements highlighting the contribution of larger or transient aggregates. ζ-potential values were close to neutrality for all samples, consistent with limited electrostatic repulsion and with the aggregation tendencies observed in some preparations. Conclusions: This study describes features of OMV produced by two relevant E. coli strains considering two isolation strategies which exert method- and strain-dependent effects on vesicle properties, including size distribution and surface charge, and emphasizes the trade-offs between yield, purity, and vesicle integrity. Full article
(This article belongs to the Special Issue Bacterial Extracellular Vesicles: Vehicles for Pathogenesis and Antibiotic Resistance)
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19 pages, 2610 KB  
Article
Evaluating Outer Membrane Vesicle Isolation Techniques for Borrelia burgdorferi and Their Impact on Vesicle Composition, Gene Expression Profile and Uptake
by Jasmine Jathan, Jay M. Pandya, Mahima Jain, Tejasri Kaithalapuram, Dhara Cherukuri and Eva Sapi
Antibiotics 2025, 14(11), 1079; https://doi.org/10.3390/antibiotics14111079 - 27 Oct 2025
Cited by 2 | Viewed by 1873
Abstract
Background: Borrelia burgdorferi, the causative agent of Lyme disease, releases outer membrane vesicles (OMVs) that may contribute to infection and modulate the host immune response. Although interest in OMVs is growing, few studies have systematically compared methods for isolating OMVs from [...] Read more.
Background: Borrelia burgdorferi, the causative agent of Lyme disease, releases outer membrane vesicles (OMVs) that may contribute to infection and modulate the host immune response. Although interest in OMVs is growing, few studies have systematically compared methods for isolating OMVs from B. burgdorferi. Methods: In this study, we evaluated two OMV isolation techniques—standard ultracentrifugation and an ion-exchange chromatography-based ExoBacteria™ kit—and examined how serum supplements (rabbit serum vs. exosome-depleted fetal bovine serum, ED-FBS) influence Bb-OMV yield and composition. Gene expression profiles were assessed using RT-PCR, and specific protein content was identified by Western blot analyses. To assess the ability of Bb-OMVs to interact with host cells, Bb-OMVs were co-cultured with MDA-MB-231 triple-negative breast cancer cells. Results: Transmission electron microscopy confirmed that both methods produced spherical Bb-OMVs with intact membrane bilayers. Ultracentrifugation generated larger vesicles (15–180 nm), while the ExoBacteria™ kit yielded smaller vesicles (<50 nm) with a higher double-stranded DNA (dsDNA) content, and protein levels were similar across samples. Cultures grown with rabbit serum produced more Bb-OMVs and had cleaner backgrounds in the TEM images than those grown with ED-FBS. All Bb-OMV samples lacked intracellular markers (DnaK and 16S rRNA) and consistently expressed the outer surface protein OspA, confirming high purity. All isolated Bb-OMVs were taken up by the cells, as indicated by OspA expression, without detectable 16S rRNA, confirming vesicle internalization without bacterial contamination. Conclusions: These findings indicate that isolated OMVs are biologically active and capable of interacting with mammalian cells, highlighting their potential role in host–pathogen interactions and the broader relevance of OMVs in studying bacterial modulation of mammalian cell behavior. Overall, both isolation methods produced high-quality OMVs, with ultracentrifugation yielding slightly more pure vesicles, emphasizing the importance of selecting appropriate isolation methods and culture conditions for functional OMV studies. Full article
(This article belongs to the Special Issue Bacterial Extracellular Vesicles: Vehicles for Pathogenesis and Antibiotic Resistance)
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