Search Results (1,231)

Biofilms are microbial communities embedded in an extracellular matrix that facilitates their attachment to surfaces. This lifestyle provides advantages to pathogenic bacteria, including increased survival in the presence of antibiotics and an enhanced capacity to develop resistance. Once a biofilm is established, infections get difficult to eradicate and frequently become chronic. There is, therefore, an urgent need to develop novel strategies to counteract biofilm-associated antibiotic resistance. Here, we developed a method to monitor the evolution of antimicrobial resistance, aiming to evaluate novel drugs against bacterial resistance to antibiotics. We validated this methodology using an RNase chimera with antibiofilm activity and a reported ability to hinder colistin resistance in planktonic cultures of Acinetobacter baumannii (A. baumannii). We assessed the emergence of resistance in A. baumannii biofilms by repeated cycles of colistin exposure. This method not only preserves biofilm structure throughout treatment but also enables controlled induction of resistance acquisition while monitoring antimicrobial efficacy. Although the RNase enhanced the antibiotic’s activity against biofilms by reducing by 50% the effective dose, it did not prevent the emergence of colistin resistance, indicating that the protein may use distinct mechanisms against planktonic and biofilm communities. Nonetheless, our findings highlight the potential of this methodology for evaluating antibiotic-adjuvant candidates to combat antibiotic resistance in biofilms. Full article

Background/Objectives: Oleuropein is a bioactive phenolic compound from olive leaves with antimicrobial and antioxidant activity. This study aimed to develop a sprayable oral gel containing an oleuropein-rich aqueous extract and to evaluate its pharmaceutical performance antimicrobial efficacy and in ovo biological response. Methods: Oleuropein content was quantified using a validated chromatographic method. Polymeric systems were screened to select an optimized sprayable formulation. Physicochemical stability, dose uniformity, and antimicrobial activity against major cariogenic bacteria were evaluated. In ovo biological evaluation was conducted using the chick chorioallantoic membrane angiogenesis model together with histopathological examination of embryonic heart and liver tissues. Results: Oleuropein content was determined as 288.6 µg/mL in the olive leaf extract and 255.1 µg/mL in the final formulation. The optimized oral spray showed stable physicochemical properties, with pH maintained at 6.90 ± 0.02 and no relevant changes in viscosity during storage. The mean delivered dose per actuation was 0.128 ± 0.015 g, corresponding to 32.6 µg oleuropein per spray. The formulation exhibited inhibitory activity against all tested cariogenic microorganisms, with MIC values ranging from 13.3 to 170.7 µg/mL and MBC values generally two-fold higher. In the CAM assay, significant concentration- and time-dependent antiangiogenic effects were observed after 24–48 h at moderate and higher concentrations. Histopathological evaluation revealed dose-dependent acute degenerative and congestive changes in heart and liver tissues without evidence of fibrosis or steatosis. Conclusions: The oleuropein-based sprayable oral gel is a promising localized delivery system with adequate stability dose uniformity and antimicrobial efficacy. In ovo findings provide a conservative assessment of systemic exposure and support further development for oral biofilm and caries-related applications. Full article

Poly(hexamethylene biguanide) (PHMB) is a polycationic antimicrobial polymer exhibiting broad-spectrum activity against bacteria, fungi, and viruses, and is widely used in medical settings for infection prevention and control. However, the relationship between chemical structure and antimicrobial activity remains unclear. In this study, we synthesised and characterised a series of polymeric biguanides with systematically varied alkyl chain lengths to examine the effects of structural variation on physicochemical properties and antimicrobial activity. H NMR spectroscopy and FTIR confirmed successful polymerisation. Solubility measurements revealed a progressive decrease in aqueous solubility with increasing alkyl chain length, consistent with increased hydrophobicity. Dynamic light scattering indicated reversible folding and unfolding of polymer chains in aqueous solution, with stabilisation at higher concentrations. Diffusion-ordered spectroscopy was used to calculate hydrodynamic diameters and polydispersity indices. Antimicrobial assays against Staphylococcus aureus and Pseudomonas aeruginosa showed that polymers containing heptamethylene and octamethylene chains exhibited the highest antibacterial activity, whereas tetramethylene- and pentamethylene-containing polymers showed greater fungicidal activity against Candida albicans. Highly hydrophobic polymers showed increased aggregation, resulting in reduced antimicrobial efficacy. Overall, these results indicate that both charge density and alkyl chain length are key determinants of antimicrobial activity. This polymeric biguanide series provides a platform for further investigation of structure–activity relationships and mechanisms of action against pathogenic microorganisms and their biofilms. Full article

Antimicrobial resistance (AMR) remains a major clinical challenge, with Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species (ESKAPE) accounting for a substantial share of multidrug-resistant (MDR) infections worldwide. These organisms undermine antibiotic efficacy through reduced permeability, surface shielding, biofilm formation, and rapid genetic adaptation, mechanisms that primarily restrict effective exposure at infection sites. Bacteriophages, phage-derived enzymes, and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-based antimicrobials provide selective and mechanistically distinct alternatives to conventional antibiotics, but their performance in vivo is often limited by instability in physiological environments, immune neutralization, uneven tissue distribution, and insufficient access to bacteria protected by biofilms or surface-associated barriers. This narrative review examines how nanotechnology-based delivery systems can address these constraints. We first outline the delivery-relevant biological barrier characteristic of ESKAPE pathogens, then summarize the therapeutic potential and inherent limitations of whole phages, phage-derived enzymes, and CRISPR-based antimicrobials when used without formulation. Major nanotechnology platforms for antibacterial delivery are reviewed, followed by analysis of how nano-enabled systems can improve stability, localization, and persistence of these biological agents. A pathogen-aware integration framework is presented that links dominant barriers in each ESKAPE pathogen to the biological modality and nano-enabled delivery strategy most likely to enhance exposure at infection sites. Translational challenges, regulatory considerations, and emerging directions, including responsive delivery systems and personalized approaches, are also discussed. Overall, nano-enabled phage-based therapeutics represent a realistic and adaptable strategy for managing MDR ESKAPE infections. Therapeutic success depends on both continued discovery and engineering of antibacterial agents and effective delivery design. Full article

Vanadium carbide (V₂C) MXene shows great potential for addressing challenging implant-associated infections in bone regeneration due to its strong photothermal conversion efficiency. However, its photothermal efficacy is restricted to the near-infrared I (NIR-I) region due to a limited absorption range. To address this, we designed platinum nanoparticle-decorated V₂C heterostructures (Pt@V₂C) via an in situ growth method, leveraging Pt’s plasmonic and catalytic properties to extend the photoresponse to the NIR-II window. Subsequently, Pt@V₂C was integrated into poly-L-lactic acid (PLLA) to fabricate PLLA-Pt@V₂C scaffolds with photothermal antibacterial function by selective laser sintering. The optimized PLLA-Pt@V₂C scaffold achieves a record photothermal conversion efficiency (56.03% at 1064 nm), triggering simultaneous hyperthermia (>52 °C) and catalytic ·OH radical generation. In vitro studies demonstrate exceptional antibacterial efficacy against Staphylococcus aureus and Escherichia coli, achieving over 99% killing rates upon 1064 nm near-infrared irradiation. Furthermore, the scaffold demonstrated significant inhibition of biofilm formation, achieving over 90% reduction in biofilm biomass. Moreover, the scaffold demonstrated high cell viability, confirming its dual functionality of potent bactericidal activity and biocompatibility that supports tissue regeneration. This work provides a feasible strategy for combating implant-associated infections. Full article

Background: Periodontal disease is a chronic multifactorial inflammatory condition caused by dysbiosis of the dental biofilm, leading to destruction of the connective tissue attachment, alveolar bone resorption, and potentially tooth loss. Non-surgical periodontal therapy (NSPT), involving subgingival instrumentation, aims to restore periodontal health by reducing the probing pocket depth (PPD) and bleeding on probing (BOP) and by improving the clinical attachment level (CAL). The adjunctive use of chemical agents, such as sodium hypochlorite/amino acids (NaOCl) and cross-linked hyaluronic acid (xHyA) gels, has been proposed to enhance the efficacy of NSPT. Objective: This systematic review aimed to evaluate the clinical effectiveness of the subgingival application of NaOCl and xHyA gels as adjunctive therapies to NSPT in patients with periodontal disease. Materials and Methods: A comprehensive literature search was conducted in the MEDLINE (PubMed), Cochrane Library, Web of Science, and Scopus databases following PRISMA guidelines. The review was registered in PROSPERO (CRD420251074045). Randomized clinical trials (RCTs) in human subjects with a follow-up of at least 6 months were included if they assessed outcomes such as PPD, CAL, BOP, or radiographic bone loss (RBL). Studies involving the adjunctive use of NaOCl and xHyA gels were selected according to the PICOS strategy. Results: Two RCTs published between 2023 and 2024, with follow-ups ranging from 6 to 9 months and involving 48–50 patients, met the inclusion criteria. Both studies demonstrated significant improvements in clinical outcomes when sodium hypochlorite and hyaluronic acid were used adjunctively with NSPT compared to when NSPT was used alone. Sites treated with adjunctive therapy showed significantly greater reductions in PPD and greater CAL gains over time. Pocket closure rates were also markedly higher in deep sites (>7 mm) in the adjunctive group than in the control group, indicating a substantial regenerative potential and a possible reduction in the need for surgical intervention. Gingival recession exhibited more favorable recovery trends in the adjunctive group, while BOP frequency decreased in both groups without statistically significant differences. Conclusions: The adjunctive use of NaOCl and xHyA gels in non-surgical periodontal therapy significantly enhances clinical outcomes compared with the use of mechanical debridement alone. Full article

In this study, a silver-based metal–organic framework (Ag-MOF) nanozyme was synthesized for the synergistic eradication of drug-resistant Salmonella enteritidis in food matrices. Ag-MOF exhibits multiple enzyme-like activities, namely oxidase (OXD)-, peroxidase (POD)-, and superoxide dismutase (SOD)-like activities. It demonstrated excellent antibacterial and antibiofilm activities against erythromycin- and chloramphenicol-resistant S. enteritidis strains (N29 and P23). Specifically, treatment with 20 mg mL⁻¹ Ag-MOF resulted in nearly complete eradication of S. enteritidis in in vitro suspension assays, including 1 × 10⁷ CFU mL⁻¹ N29 strain and 6 × 10⁶ CFU mL⁻¹ P23 strain. Moreover, treatment with 1 mg mL⁻¹ Ag-MOF led to 80~90% biofilm inhibition of S. enteritidis. Mechanistic investigations revealed that Ag-MOF effectively interacted with amino-rich structures on the bacterial surface (such as membrane proteins and peptidoglycan components), generated abundant reactive oxygen species (ROS), released Ag⁺ ions, and depleted intracellular glutathione, which collectively disrupted cell membrane integrity and induced severe leakage of intracellular proteins and nucleic acids. Importantly, Ag-MOF maintained high antibacterial efficacy in complex simulated food matrices (pork, milk, and egg shell). Overall, this study offers key insights into enzyme-mimicking antibacterial materials and a promising strategy to combat multidrug resistant foodborne pathogens. Full article

Campylobacter jejuni is an important foodborne pathogen. To prevent human infections, special attention should be paid to prevention. Recently, methods involving essential oils have been considered as a means of reducing the number of contaminants in and on foods. This review summarizes the results of studies in which essential oils (EOs) with anti-campylobacter effects were tested. The most widely studied EOs were clove (28%), oregano (24%), thyme (22%), rosemary (8%), lavender (7%), sage (7%), and tea tree (4%), with other EOs studied to a lesser extent. The anti-Campylobacter efficacies of these EOs were demonstrated in vitro using a broad repertoire of methods, such as minimal inhibitory and bactericidal concentrations, agar diffusion, time-kill assays, adhesion and biofilm inhibitory assays, two-dimensional polyacrylamide gel electrophoresis, quantitative reverse-transcription PCR, and liquid chromatography–mass spectrometry. Recent studies have also focused on the practical application of such EOs, with experiments performed on different food matrices, typically chicken, duck, and beef. The most frequent treatment methods were mixing, dipping, and short-time freezing, either in packed or unpacked forms, and storage at different temperatures (typically 4 °C), although experiments were also performed at 25 °C, 32 °C, and 42 °C using different EO concentrations. In summary, these experiments revealed the anti-Campylobacter effects of thyme, cinnamon, coriander, lime, oregano, chrysanthemum, and basil. Full article

Chronic Pseudomonas aeruginosa lung infections in cystic fibrosis (CF) represent one of the most treatment-refractory bacterial diseases, sustained by biofilm formation, metabolic dormancy, and adaptive antibiotic resistance evolution. While bacteriophage (phage) therapy has emerged as a promising alternative for multidrug-resistant (MDR) pathogens, clinical studies in CF have demonstrated transient reductions in bacterial burden without achieving complete eradication. This review integrates molecular, evolutionary, and immunological findings to explain the multifactorial barriers that limit phage therapeutic efficacy in chronic CF infections. We highlight three major obstacles: (i) bacterial dormancy and persistence within biofilms that restrict phage adsorption and replication; (ii) hypermutability and extensive genotypic diversification of CF-adapted P. aeruginosa, which accelerate phage resistance evolution and necessitate broad host-range coverage; and (iii) CF-specific immune constraints—including a dysfunctional innate immune system and phage-neutralizing humoral immunity—that reduce phage bioavailability and undermine sustained bacterial clearance. Emerging strategies to overcome these challenges include the discovery of dormant-targeting phages capable of replicating in metabolically quiescent cells, evolution-informed phage training to delay resistance evolution, and synthetic phage engineering approaches designed to disrupt biofilms and expand host-range coverage. In parallel, computational or artificial intelligence (AI)-guided frameworks for phage cocktail design and cystic fibrosis transmembrane conductance regulator (CFTR) modulator-mediated restoration of host immune function together offer a more integrated therapeutic paradigm that unites phage biology and host immune context. By unifying clinical outcomes with mechanistic, evolutionary, and immunological perspectives, this review outlines a next-generation framework for phage therapy in CF aimed at achieving more durable therapeutic outcomes. Full article

Lower respiratory tract infections caused by Pseudomonas aeruginosa are a global concern. Patients with chronic lung diseases such as cystic fibrosis and non-cystic fibrosis bronchiectasis often do not receive adequate antibiotic delivery through conventional routes. P. aeruginosa employs several mechanisms, including biofilm formation and efflux pumps to limit the accumulation of bactericidal drug concentrations. Direct drug delivery to the lung epithelial lining fluid can increase antibiotic concentration and reduce treatment failure rates. This review discusses current research and developments in inhaled antibiotic formulations for treating P. aeruginosa infections. Recent studies on particle engineering for the dry powder inhalers of antibiotics emphasized three fundamental principles of development: micro, nano, and nano-in-microparticles. Carrier-free microparticles showed potential for high-dose delivery but suffered from poor aerosolization, which could be improved through a drug–drug combination. Amino acids in a co-spray-dried system improved powders’ aerodynamics and reduced moisture sensitivity while incorporating the chitosan/poly(lactic-co-glycolic acid) (PLGA)-modified release of the drug. Nano-in-microsystems, embedding lipid carriers, showed improved antibiofilm activity and controlled release. We also highlight emerging biologics, including antibacterial proteins/peptides, vaccines, bacteriophages, and probiotics. Research on antibiotics and biologics for inhalation suggests excellent safety profiles and encouraging efficacy for some formulations, including antimicrobial peptides and bacteriophage formulations. Further research on novel molecules and synergistic biologic combinations, supported by comprehensive animal lung safety investigations, will be required in future developments. Full article

Background/Objectives: Wound infections caused by multidrug-resistant Gram-negative bacteria, such as Escherichia coli, Pseudomonas aeruginosa, and Acinetobacter baumannii, pose a major clinical challenge. This study evaluated the interactions between gamma-polyglutamic acid (γ-PGA), cefiderocol, and silver nanoparticles (AgNPs) within multilayer wound dressing configurations. The primary goal was to clarify the dual role of γ-PGA as a healing promoter and a potential protector of bacterial cells against antimicrobial agents. Methods: Multilayer dressing models were assembled in 96-well plates to simulate vertical stratification of antimicrobial layers4. Bacterial viability was assessed through relative OD₆₀₀ measurements following incubation with varying concentrations and spatial arrangements of cefiderocol, AgNPs, and γ-PGA. Data were analyzed using generalized linear modeling (GLM) with a gamma distribution and random forest regression to determine the relative importance of each factor in modulating bacterial survival. Results: γ-PGA concentration emerged as the dominant factor influencing bacterial viability, accounting for nearly 100% of variable importance in random forest analysis. Despite high antimicrobial pressure from cefiderocol and AgNPs, bacterial viability stabilized at approximately 40% in the presence of γ-PGA. The vertical positioning of γ-PGA significantly impacted survival; direct physical contact between the polymer and bacteria, particularly at high concentrations, enhanced bacterial persistence in P. aeruginosa and E. coli. Cefiderocol showed strain-specific potency, while AgNPs provided consistent growth inhibition. Conclusions: γ-PGA plays a paradoxical role in wound care by providing moisture retention while simultaneously acting as a cytoprotective agent that reduces antimicrobial efficacy, likely by facilitating biofilm formation. These findings underscore the necessity of optimizing the spatial layering and concentration of biopolymers in advanced dressings. Strategic design is crucial to balance regenerative benefits with maximal antimicrobial control to improve clinical outcomes in chronic wound management. Full article

This in vitro study compared the antimicrobial and antibiofilm efficacy of four commercially available chlorhexidine (CHX)-based mouthwashes, with different nominal CHX concentrations, against clinically relevant postoperative oral pathogens, including Staphylococcus aureus, Streptococcus mutans, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, and Candida auris. Antimicrobial potency was evaluated using MIC and CEMIC indices, while biofilm thickness reduction was quantified using 3D digital microscopy and custom image analysis software. Among the tested formulations, the excipient-enriched formulation exhibited the lowest MIC values and the most significant reduction in biofilm thickness, particularly against Gram-negative bacteria and Candida species. All mouthwashes achieved CEMIC < 0.1, confirming high theoretical applicability margins; however, CEMIC reflects potential clinical usefulness rather than clinical superiority. The findings demonstrate that the antimicrobial and antibiofilm activity of CHX rinses is formulation-dependent and cannot be predicted solely by CHX concentration. The influence of excipients is discussed as a possible contributing factor, but related mechanisms remain speculative and require direct validation in future studies. This work supports a formulation-driven, evidence-based approach to antiseptic comparison in postoperative dentistry, without assessing clinical wound-healing outcomes. Full article

This review focuses on the research progress on natural products as β-lactam antibiotic adjuvants, aiming to address the escalating challenge of antibiotic resistance, particularly the inactivation of antibiotics caused by β-lactamases. The article provides an in-depth analysis of the mechanisms by which plant-derived (e.g., flavonoids, tannins, phenolics, terpenoids, and alkaloids) and microbial-derived (e.g., clavulanic acid, fungal metabolites, bacteriophages) natural products enhance antimicrobial efficacy. Key potentiation strategies discussed include efflux pump inhibition, membrane permeability alteration, biofilm disruption, PBP2a inhibition, and direct β-lactamase inhibition. Additionally, the review outlines in vitro methods (e.g., dilution and checkerboard assays) and in vivo models (e.g., mouse infection models) used to assess synergistic effects. It also addresses major challenges in identifying active compounds, elucidating mechanisms of action, and pharmacokinetic characterization. Looking forward, the article highlights the potential of multi-omics approaches, artificial intelligence, and nanotechnology to overcome existing bottlenecks, providing novel strategies for the development of effective and safe antibiotic adjuvants. These advances are expected to provide both theoretical insights and practical guidance for combating antibiotic-resistant bacterial infections. Full article

The valorization of agri-food residues is crucial for advancing circular bioeconomy strategies and mitigating environmental impacts. Turnip greens (Brassica rapa subsp. sylvestris) are a traditional vegetable cultivated in southern Italy. While the edible portions include flower sprouts, buds, and young leaves, the more leathery leaves and stems are typically discarded. These wastes represent valuable sources of compounds with antioxidant and antimicrobial potential. This study aims to develop the extraction of phenolic compounds from turnip green residues using two techniques: silent maceration and ultrasound-assisted extraction (UAE). Ethanol was selected over methanol as a food-safe alternative solvent, with preliminary tests confirming equivalent efficiency. A Design of Experiments (DoE) approach was applied to both leaves and stems to assess the effects of solvent composition, solvent-to-matrix ratio, and extraction time on Total Phenolic Content and Trolox Equivalent Antioxidant Capacity. DoE results identified UAE as the most effective method for stems, while for leaves, the solvent-to-dry-mass ratio was the key parameter. HPLC-DAD analysis was performed to identify and quantify the phenolic acids in selected extracts. The antibacterial activity of these extracts against biofilms of six pathogenic strains was evaluated using crystal violet and MTT assays, confirming efficacy in both biofilm formation and mature stages. Full article

Due to the limitations of conventional antibiotics, antimicrobial peptides (AMPs) have emerged as promising therapeutic alternatives for the prevention and treatment of oral infections. This study systematically evaluated in vitro evidence regarding the antimicrobial and anti-biofilm activity of natural AMPs against oral pathogens. A systematic search using the PICOT strategy was conducted in PubMed, EMBASE, and Scopus, retrieving 7711 articles. After title and abstract screening, 109 studies were selected for full-text analysis, resulting in 26 articles that met the eligibility criteria. Among the AMPs evaluated, nisin (n = 15) and LL-37 (n = 5) were the most frequently investigated, while other peptides included lactoferrin, lactoferricin, melittin, lysozyme, histatin-5, cystatin C, chromogranin A, parasin-1, protamine, AmyI-1-18, and DCD-1L. Natural AMPs of human and animal origin demonstrated antimicrobial activity against bacteria associated with oral infections, particularly Streptococcus mutans and Enterococcus faecalis. These peptides were tested in different formulations, including solutions, incorporation into dental materials and polymers, and application in sonodynamic antimicrobial therapy. Overall, the findings indicate that natural AMPs represent a promising class of biomolecules for controlling oral biofilms; however, further clinical studies are required to validate their long-term efficacy and safety. Full article