Wound Infection Model Research Articles

Biofilm Microenvironment-Sensitive Anti-Virulent and Immunomodulatory Nano-on-Nanodroplets to Combat Refractory Biofilm Infection Through Toxin Neutralization and Phagocytosis.

Biofilm-associated wound infection is principally perceived as the bacterial defense mechanism that hinders antibiotic penetration, causes toxin impairment, and suppresses the immunological responses of the host immune system. Several antibiofilm agents have been developed, but the least of these agents can simultaneously cornerstone on the biofilm-associated immunosuppression and bacterial toxin-induced cellular dysfunction. Inspired by the fusogenic property of nanodroplets and immunomodulatory functions of metal nanoparticles, biofilm targeted anti-virulent immunomodulatory cationic nanoparticle shelled nanodroplets (C-AgND) is fabricated to completely disintegrate and eradicate the Staphylococcus aureus (S. aureus) biofilm. The specific binding of C-AgND neutralizes the negatively charged EPS layer, causing their destabilization followed by penetration of the nanoformulation into the biofilm matrix, killing the persister cells. Consequently, C-AgND eliminates the virulence property of the S. aureus biofilm through α-hemolysin neutralization. C-AgND promotes a strong immunomodulatory effect by polarizing macrophages into their M1 phenotype to induce phagocytosis of the disintegrated biofilm-released residual cells, rejuvenating the host's innate immune responses for the complete eradication of the biofilm. Moreover, the ex vivo skin wound infection model illustrates an excellent biofilm eradication efficacy of C-AgND in comparison to the commercial ones, rendering them to be a promising replacement of existing antibiofilm agents in clinical application.

Radical-Scavenging Violet Phosphorus Nanosheets for Attenuating Hyperinflammation and Promoting Infected Wound Healing.

Antibacterial therapy targeting the regulation of macrophage polarization may be a useful approach for normalizing the immune environment and accelerating wound healing. Inspired by black phosphorus-based nanoplatforms, more stable yet less-explored violet phosphorus nanosheets (VPNSs) are expected to provide a superior solution for effectively combating bacterial infections. In this study, an average thickness of 5-7nm VPNSs arefabricated through the liquid-phase exfoliation method to serve as an immunoregulatory dressing for the treatment of infected wounds. VPNSs attenuated excessive reactive oxygen species (ROS) and reduced the accumulation of proinflammatory M1 macrophages, showing notable antioxidant and anti-inflammatory properties. Comprehensive RNA sequencing further elucidated the potential immunoregulatory mechanisms of VPNSs, including modulation of the inflammatory response and enzyme regulator activity. Additionally, the inherent photothermal properties of the VPNSs contributed significantly to their antibacterial efficacy. When combined with near-infrared laser irradiation, VPNSs showed remarkable effectiveness in reducing infection-related complications and expediting wound healing in infected skin wound models. The rapid promotion of wound healing through ROS clearance, the regulation of macrophage polarization, and hyperthermia generation underscores the potential of the violet-phosphorus-based nanoplatforms as clinically viable agents for treating infected wounds. This study suggests that VPNSs are promising candidates for clinical anti-infective and anti-inflammatory applications.

Recombinant Keratin-Chitosan Cryogel Decorated with Gallic Acid-Reduced Silver Nanoparticles for Wound Healing.

Wound healing is a complex physiological process that can be roughly divided into four stages: hemostasis, inflammation, proliferation, and remodeling. Conventional wound dressings often fail to meet the diverse needs of these healing stages due to their limited functionality. Cryogels, however, possess several attractive properties, such as large, interconnected pores, good mechanical strength, and ease of modification, making them suitable for developing advanced dressings with multiple functions. In this study, we developed a multifunctional cryogel dressing, with biocompatible polysaccharides as the main component, designed to provide a breathable, moist, and antibacterial microenvironment for chronic infected wounds, thereby promoting wound healing. Recombinant keratin 31 (RK31) was combined with chitosan (CS) to produce a CS/RK31 cryogel, referred to as CK. Gallic acid-reduced silver nanoparticles (GA/Ag NPs) were incorporated as the active antibacterial component to create the CS/K31@GA/Ag cryogel, known as CKGA. The cryogel was characterized using scanning electron microscopy (SEM) and a universal testing machine, and its biocompatibility was assessed in vitro. The dynamic hemostatic performance of the cryogel was evaluated with a rat tail amputation bleeding model. Additionally, the antibacterial effects of the cryogel against Staphylococcus aureus and Escherichia coli were tested using agar diffusion assays and turbidimetry. The antioxidant capacity of the CKGA cryogel was also measured in vitro. Finally, the cryogel's ability to promote wound healing was tested in an SD rat model of infected wounds. Characterization results showed that the CKGA cryogel features an interpenetrating porous network structure and exhibits excellent mechanical properties, with a swelling rate of up to 1800%. Both in vitro and in vivo experiments confirmed that the cryogel has good biocompatibility, effectively absorbs exudates, and rapidly stops bleeding. The addition of GA/Ag NPs provided significant antibacterial effects, achieving an inhibition rate of over 99.9% against both S. aureus and E. coli. Furthermore, CKGA cryogels demonstrated a strong scavenging capacity for ROS in a dose-dependent manner. Studies using the SD rat infected wound model showed that the cryogel effectively inhibited bacterial proliferation on wound surfaces, reduced local tissue inflammation, and promoted the healing of infected wounds. The multifunctional cryogel, with its rapid hemostatic, antibacterial, and antioxidant properties, as well as its ability to promote cell proliferation, could be widely used as a wound dressing for the healing of bacterial infections.

Novel antibacterial pullulan derivatives modified with quaternary phosphonium salts for infected wound treatments

The development of novel antibacterial agents is urgently needed to tackle bacterial infection, the major global issue menacing human health. Among them, polymeric quaternary phosphonium salts are worth noticing owing to their strong antibacterial activities and other merits including low bacterial drug resistance. Herein, pullulan modified with quaternary phosphonium salts (PQP) was synthesized using esterification reactions of pullulan and (5-carboxypentyl)triphenylphosphonium bromide (CPTPPB) mediated by N,N′‑carbonyldiimidazole (CDI), and was evaluated as novel antibacterial agents for treating wound infection for the first time. The chemical structures, chemical bonding, elemental compositions, crystalline properties and thermostability of PQP were systematically investigated. PQP exhibited in vitro antibacterial activities against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli), which were unveiled by the spread plate method and possibly resulted from the damage of bacterial cell walls/membranes mediated by electrostatic and hydrophobic interactions according to preliminary mechanism studies. Weak cytotoxicity and excellent hemocompatibility of PQP at the effective bactericidal concentration were observed. Moreover, in the infected wound model of mice, PQP was capable of disinfecting the wound and accelerating the healing. We opine PQP in this work is promising for antibacterial applications and will inspire the synthesis of novel antibacterial agents derived from natural polymers.

A pH-modulated polymeric drug-release system simultaneously combats biofilms and planktonic bacteria to eliminate bacterial biofilm

Bacterial infection, leading to biofilm formation, raises grave concerns. With the aim of eliminating both integrity of biofilms and individual bacterial cells, it is desirable to develop bacterial biofilm-eliminating material to be combined with dispersal and anti-microbial agents. Herein, a biofilm-eliminating enzyme (α amylase) was incorporated in novel pH-sensitive polymer hydrogel, Poly (methacrylic acid-co-2-(dimethylamino)ethyl methacrylate), which encapsulates drug-loaded porous Zeolitic Imidazolate Framework-8 (ZIF-8). At body temperature (37 °C) the maximum swelling of the hydrogel was observed at pH 8.5, which assisted in rupturing biofilm by releasing α amylase. Amylase, encapsulated in polymer matrix reached 90% release after 6 days at 8.5, while CIP was released 96% on day 23 at pH 8.5. The antibiofilm activity of drug-releasing system was assessed against E. coli and S. aureus. The overall bacterial viability of biofilms showed significant time-dependent declines, when exposed to drug-loaded hydrogel. Importantly, cytotoxicity experiments showed excellent biocompatibility of hydrogel. In the in vivo deep wound infection model, drug-loaded hydrogel treatment showed complete wound closure with proper tissue regeneration. No inflammatory, or systemic toxicity of hydrogel was observed. The drug release from hydrogel is initiated only upon receiving physiological cue arising due to biofilm formation. Therefore, this hydrogel serves as reservoir for antibacterial drug, which release it only in presence of biofilm and thus work for sustained release for long period, compared to conventional drug loaded films or coatings. A powerful modality for management of biofilm-associated infections can be provided by the proposed synergistic strategy for complete elimination of biofilms.

Dual action electrochemical bandage operated by a programmable multimodal wearable potentiostat

We have developed electrochemical bandage (e-bandage) prototypes that generate the reactive oxygen species hypochlorous acid (HOCl) or hydrogen peroxide (H2O2) for potential use to treat biofilm-infected wounds in humans. We have shown that both e-bandage-generated HOCl and H2O2 kill biofilms in vitro and in infected wounds on mice, with the former being more active in vitro. The H2O2-generating e-bandage, more so than the HOCl-generating e-bandage, was associated with improved healing of infected wounds. Here, a strategy in which H2O2 and HOCl are alternately generated—for dual action—was explored. The goal was to develop a programmable multimodal wearable potentiostat [PMWP] that can generate HOCl or H2O2, as needed. An ultralow-power microcontroller unit was developed to manage operation of the PMWP. The system was operated with a 260-mAh capacity coin battery and weighed 4.6 g, making it suitable for future small animal experiments (and ultimately, potential evaluation in humans). As assessed using electrochemical parameters, the device functioned comparably to a commercial benchtop potentiostat. To confirm antimicrobial activity, PMWP-controlled e-bandages were tested in vitro against clinical isolates of methicillin-resistant Staphylococcus aureus, Pseudomonas aeruginosa, Acinetobacter baumannii, Enterococcus faecium, and Candida auris. When programmed to deliver HOCl followed by H2O2, PMWP-controlled e-bandages exhibited activity against biofilms of all study isolates tested. Finally, we demonstrated the PMWP's usability in a murine wound infection model.

Screening of the Pandemic Response Box library identified promising compound candidate drug combinations against extensively drug-resistant Acinetobacter baumannii

Infections caused by antimicrobial-resistant Acinetobacter baumannii pose a significant threat to human health, particularly in the context of hospital-acquired infections. As existing antibiotics lose efficacy against Acinetobacter isolates, there is an urgent need for the development of novel antimicrobial agents. In this study, we assessed 400 structurally diverse compounds from the Medicines for Malaria Pandemic Response Box for their activity against two clinical isolates of A. baumannii: A. baumannii 5075, known for its extensive drug resistance, and A. baumannii QS17-1084, obtained from an infected wound in a Thai patient. Among the compounds tested, seven from the Pathogen box exhibited inhibitory effects on the in vitro growth of A. baumannii isolates, with IC50s ≤ 48 µM for A. baumannii QS17-1084 and IC50s ≤ 17 µM for A. baumannii 5075. Notably, two of these compounds, MUT056399 and MMV1580854, shared chemical scaffolds resembling triclosan. Further investigations involving drug combinations identified five synergistic drug combinations, suggesting potential avenues for therapeutic development. The combination of MUT056399 and brilacidin against A. baumannii QS17-1084 and that of MUT056399 and eravacycline against A. baumannii 5075 showed bactericidal activity. These combinations significantly inhibited biofilm formation produced by both A. baumannii strains. Our findings highlight the drug combinations as promising candidates for further evaluation in murine wound infection models against multidrug-resistant A. baumannii. These compounds hold potential for addressing the critical need for effective antibiotics in the face of rising antimicrobial resistance.

Multifunctional EGCG@ZIF-8 Nanoplatform with Photodynamic Therapy/Chemodynamic Therapy Antibacterial Properties Promotes Infected Wound Healing.

Damaged skin is susceptible to invasion by harmful microorganisms, especially Staphylococcus aureus and Escherichia coli, which can delay healing. Epigallocatechin-3-gallate (EGCG) is a natural compound known for effectively promoting wound healing and its potent anti-inflammatory effects. However, its application is limited due to its susceptibility to oxidation and isomerization, which alter its structure. The use of zeolitic imidazolate framework-8 (ZIF-8) can effectively tackle these issues. This study introduces an oxygen (O2) and hydrogen peroxide (H2O2) self-supplying ZIF-8 nanoplatform designed to enhance the bioavailability of EGCG, combining photodynamic therapy (PDT) and chemodynamic therapy (CDT) to improve antibacterial properties and ultimately accelerate wound healing. For this purpose, EGCG and indocyanine green (ICG), a photosensitizer, were successively integrated into a ZIF-8, and coated with bovine serum albumin (BSA) to enhance biocompatibility. The outer layer of this construct was further modified with manganese dioxide (MnO2) to promote CDT and calcium peroxide (CaO2) to supply H2O2 and O2, resulting in the final nanoplatform EGCG-ICG@ZIF-8/BSA-MnO2/CaO2 (EIZBMC). In in vitro experiments under 808 nm laser, EIZBMC exhibited synergistic antibacterial effects through PDT and CDT. This combination effectively released reactive oxygen species (ROS), which mediated oxidative stress to inhibit the bacteria. Subsequently, in a murine model of wound infection, EIZBMC not only exerted antibacterial effects through PDT and CDT but also alleviated the inflammatory condition and promoted the regeneration of collagen fibers, which led to accelerated wound healing. Overall, this research presents a promising approach to enhancing the therapeutic efficacy of EGCG by leveraging the synergistic antibacterial effects of PDT and CDT. This multifunctional nanoplatform maximizes EGCG's anti-inflammatory properties, offering a potent solution for promoting infected wound healing.

Three in One with Dual-Functional Hydrogel of Lactoferrin/NZ2114/LMSH Promoting Staphylococcus aureus-Infected Wound Healing.

Wound infections caused by Staphylococcus aureus often result in localized suppurative lesions that severely impede the healing process, so it is urgent to develop a dress with efficient antimicrobial and pro-healing functions. In this study, the bifunctional injectable hydrogel lactoferrin (Lf)/NZ2114/lithium magnesium silicate hydrogel (LMSH) was first successfully prepared through the electrostatic interaction method. The physical, biological, and efficacy properties are systematically analyzed with good shear-thinning capacity and biocompatibility. More importantly, it inhibits infection and promotes wound healing in a mouse wound infection model after 14 d treatment, and the bactericidal rate and healing rate were over 99.92% and nearly 100%, respectively. Meanwhile, the massive reduction of inflammatory cells, restoration of tissue structure, and angiogenesis in mice showed the anti-inflammatory and pro-healing properties of the hydrogel. The healed wounds showed thickening with more hair follicles and glands, suggesting that the hydrogel Lf/NZ2114/LMSH (Three in One) could be a better dressing candidate for the treatment of S. aureus-induced wound infections.

PH-responsive mupirocin-loaded hybrid nanoparticles in hydrogel and film forming spray against resistant bacterial wound infections

The aim of this work was to fabricate smart Mupirocin loaded Lipid Polymeric Hybrid Nanoparticles (MUP-LPHNs) based hydrogel and Film forming spray (FFS) for targeted release of MUP in response to wound pH. The MUP-LPHNs were optimized through design expert v.12. The optimized MUP-LPHNs had a particle size of 270.7 ± 3.20 nm with zeta potential of 17.05 ± 1.20 mV and entrapment efficiency of 88.0 ± 0.35 %. MUP-LPHNs showed no major chemical interaction, spherical morphology and amorphous nature as confirmed by FTIR, SEM and XRD, respectively. MUP-LPHNs were loaded to MUP-LPHNs based hydrogel (MUP-LPHNs-HG) and MUP-LPHNs based film forming spray (MUP-LPHNs-FFS) with successful characterization. In vitro release data confirmed sustained and pH dependent release from both MUP-LPHNs-HG and MUP-LPHNs-FFS as higher release was observed at pH 7.4. MUP-LPHNs-HG and MUP-LPHNs-FFS showed high efficiency and long-term inhibition of Staphylococcus aureus (SA), Methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa (P. aeruginosa) with 4–6 fold increase in the antibacterial activity compared to MUP solution. MUP-LPHNs-HG and MUP-LPHNs-FFS exhibited both in vitro and in vivo wound pH contingent, sustained drug release. Moreover, the prepared nanocarriers systems flaunted significant therapeutic efficacy against MRSA infected wound model by significant reduction in bacterial load and wound area, increased hydroxy proline level and antioxidant enzymes and reduced proinflammatory cytokines. Similarly, immunohistochemistry, histopathological and trichrome assessment showed significant reduction in level of inflammatory markers, accelerated and enhanced re-epithelization, improved connective tissue remodeling and effectively promote collagen deposition with effective wound regeneration, respectively. It can be concluded that LPHNs could be a potential carrier system for enhancing the therapeutic efficacy of MUP against resistant bacterial wound infections.

Dual-species proteomics and targeted intervention of animal-pathogen interactions

IntroductionHost-microbe interactions are important to human health and ecosystems globally, so elucidating the complex host-microbe interactions and associated protein expressions drives the need to develop sensitive and accurate biochemical techniques. Current proteomics techniques reveal information from the point of view of either the host or microbe, but do not provide data on the corresponding partner. Moreover, it remains challenging to simultaneously study host-microbe proteomes that reflect the direct competition between host and microbe. This raises the need to develop a dual-species proteomics method for host-microbe interactions. ObjectivesWe aim to establish a forward + reverse Stable Isotope Labeling with Amino acids in Cell culture (SILAC) proteomics approach to simultaneously label and quantify newly-expressed proteins of host and microbe without physical isolation, for investigating mechanisms in direct host-microbe interactions. MethodsUsing Caenorhabditis elegans-Pseudomonas aeruginosa infection model as proof-of-concept, we employed SILAC proteomics and molecular pathway analysis to characterize the differentially-expressed microbial and host proteins. We then used molecular docking and chemical characterization to identify chemical inhibitors that intercept host-microbe interactions and eliminate microbial infection. ResultsBased on our proteomics results, we studied the iron competition between pathogen iron scavenger and host iron uptake protein, where P. aeruginosa upregulated pyoverdine synthesis protein (PvdA) (fold-change of 5.2313) and secreted pyoverdine, and C. elegans expressed ferritin (FTN-2) (fold-change of 3.4057). Targeted intervention of iron competition was achieved using Galangin, a ginger-derived phytochemical that inhibited pyoverdine production and biofilm formation in P. aeruginosa. The Galangin-ciprofloxacin combinatorial therapy could eliminate P. aeruginosa biofilms in a fish wound infection model, and enabled animal survival. ConclusionOur work provides a novel SILAC-based proteomics method that can simultaneously evaluate host and microbe proteomes, with future applications in higher host organisms and other microbial species. It also provides insights into the mechanisms dictating host-microbe interactions, offering novel strategies for anti-infective therapy.

Comparison of St. John's wort oil and thiocilline ointment on wound healing in a diabetic rat model

Aims: In diabetic patients, wound healing is impaired and wounds are often infected with multifactorial agents. This study aimed to compare the effectiveness of St. John’s wort and ointment containing bacitracin-neomycin (thiocilline) to improve wound healing in a diabetic infected wound model. Methods: Rats in which diabetes was induced by administering 60 mg/kg streptozotocin (STZ) were considered diabetic if their blood sugar levels were above 300 mg/dl 72 hours later. Group 1: Control (Non diabetic) group, Group 2: Diabetes group. During wound care, both groups were disinfected with povidone iodine (PI) and the right lumbar region of each rat was dressed with thiocilline and the left lumbar region was dressed with St. John’s wort oil. Considering the wound healing period, the study was terminated after an average of 20 days. In histopathological examination, ulceration, necrosis, epithelialization, congestion, edema, polymorphous nucleated leukocyte (PNL), monocyte, fibroblast, and neovascularization were evaluated. Results: In histopathological evaluation, there was a statistically significant decrease in ulceration and necrosis in the group treated with St. John’s wort oil compared to the group given thiocilline (p=0.04). In terms of epithelialization, there was a statistically significant increase in the group dressed with St. John’s wort oil compared to the group given thiocilline (p=0.03). There was a statistically significant decrease in congestion and edema in the group treated with St. John’s wort oil compared to the group given thiocilline (p=0.03). There was a statistically significant increase in fibroblast and neovascularization in the group treated with St. John’s wort oil compared to the group given thiocilline (p=0.02). Conclusion: Histopathologic ally, epithelialization, fibroblast, and neovascularization, which have important functions in the wound healing process, increased in diabetic rats administered St. John’s wort. Although it is used in traditional medicine due to its antidepressant effectiveness, we believe that St. John’s wort can be used in wounds that develop in diabetic patients, as it has the potential to increase the wound healing process.

Optimized Dosing and Delivery of Bacteriophage Therapy for Wound Infections.

Lytic bacteriophages, viruses that lyse (kill) bacteria, hold great promise for treating infections, including wound infections caused by antimicrobial-resistant Pseudomonas aeruginosa. However, the optimal dosing and delivery strategies for phage therapy remain unclear. In a mouse wound infection model, we investigated the impact of dose, frequency, and administration route on the efficacy of phage therapy. We find that topical but not intravenous delivery is effective in this model. High-doses of phage reduces bacterial burden more effectively than low-doses, and repeated dosing achieves the highest eradication rates. Building on these insights, we developed "HydroPhage", a hyaluronan-based hydrogel system that uses dynamic covalent crosslinking to deliver high-titre phages over one week. HydroPhage eradicates infections five times more effectively than intravenous injection. We conclude that hydrogel-based sustained phage delivery enhances the efficacy of phage therapy and offers a practical, well-tolerated option for topical application.

Caffeic Acid and Cyclen-Based Hydrogel for Synergistic Antibacterial Therapy.

Caffeic acid is a natural product that contains both phenolic and acrylic functional groups and has been widely employed as an alternative drug to combat chronic infections induced by microbes such as bacteria, fungi, and viruses. Several strategies, including derivatization and nanoformulation, have been applied in order to overcome the issues of water insolubility, poor stability, and the bioavailability of caffeic acid. Here, caffeic acid and cyclen-Zn(II) are incorporated into a G4-assembly by using a phenylborate linker to form the mixed supramolecular prodrug GB-CA/Cy-Zn(II) hydrogel. The delivery system is expected to enhance antibacterial and anti-inflammatory properties during the wound healing process through the synergistic effect of caffeic acid and cyclen-Zn(II). The preparation and physicochemical and mechanical properties of the hydrogel were investigated by NMR, CD, TEM, and rheological assays. The typical inflammatory cytokines and in vitro antibacterial experiments indicated that inflammation and infection can be significant suppressed by the hydrogel treatment. An in vivo infected wound model treated by the hydrogel showed rapid wound healing capacity and biosafety. The current work depicts a simple method to prepare a caffeic acid hydrogel carrier, which facilitates synergistic treatment for inflammation and bacterial infections at the wound site.

Dual quorum-sensing control of purine biosynthesis drives pathogenic fitness of Enterococcus faecalis.

Enterococcus faecalis is a resident of the human gut, though upon translocation to the blood or body tissues, it can be pathogenic. Here we discover and characterize two peptide-based quorum-sensing systems that transcriptionally modulate de novo purine biosynthesis in E. faecalis. Using a comparative genomic analysis, we find that most enterococcal species do not encode this system; E. moraviensis, E. haemoperoxidus and E. caccae, three species that are closely related to E. faecalis, encode one of the two systems, and only E. faecalis encodes both systems. We show that these systems are important for the intracellular survival of E. faecalis within macrophages and for the fitness of E. faecalis in a murine wound infection model. Taken together, we combine comparative genomics, microbiological, bacterial genetics, transcriptomics, targeted proteomics and animal model experiments to describe a paired quorum sensing mechanism that directly influences central metabolism and impacts the pathogenicity of E. faecalis.

ANTIMICROBIAL WOUND DRESSINGS FOR FULL-THICKNESS INFECTED BURN WOUNDS.

Infection of wounds delays healing, increases treatment costs, and leads to major complications. Current methods to manage such infections include antibiotic ointments and antimicrobial wound dressings, both of which have significant drawbacks, including frequent reapplication and contribution to antimicrobial resistance. In this work, we developed wound dressings fabricated with a medical-grade polyurethane coating composed of natural plant secondary metabolites, cinnamaldehyde, and alpha-terpineol. Our wound dressings are easy to change and do not adhere to the wound bed. They kill gram-positive and -negative microbes in infected wounds due to the Food and Drug Administration-approved for human consumption components. The wound dressings were fabricated by dip coating. Antimicrobial efficacy was determined by quantifying the bacteria colonies after a 24 h of immersion. Wound healing and bacterial reduction were assessed in an in vivo full-thickness porcine burn model. Our antimicrobial wound dressings showed a > 5-log reduction (99.999%) of different gram-positive and gram-negative bacteria, while maintaining absorbency. In the in vivo porcine burn model, our wound dressings were superior to bacitracin in decreasing bacterial burden during daily changes, without interfering with wound healing. Additionally, the dressings had a significantly lower adhesion to the wound bed. Our antimicrobial wound dressings reduced the burden of clinically relevant bacteria more than commercial antimicrobial wound dressings. In an in vivo infected burn wound model, our coatings performed as well or better than bacitracin. We anticipate that our wound dressings would be useful for the treatment of various types of acute and chronic wounds.

Discovery of quaternized pyridine-thiazole-ruthenium complexes as potent anti-Staphylococcus aureus agents

Quaternization of ruthenium complexes may be a promising strategy for the development of new antibiotics. In response to the increasing bacterial resistance, we integrated the quaternary amine structure into the design of ruthenium complexes and evaluated their antibacterial activity. All the ruthenium complexes showed good antibacterial activity against the tested Staphylococcus aureus (S. aureus). Ru-8 was the most effective antibacterial agent that displayed excellent antibacterial activity against S. aureus (MIC = 0.78–1.56 μg/mL). In vitro experiments showed that all nine ruthenium complexes had low hemolytic toxicity to rabbit erythrocytes. Notably, Ru-8 was found to disrupt bacterial cell membranes, alter their permeability, and induce ROS production in bacteria, all the above leading to the death of bacteria without inducing drug resistance. To further explore the antibacterial activity of Ru-8in vivo, we established a mouse skin wound infection model and a G. mellonella larvae infection model. Ru-8 exhibited significant antibacterial efficacy against S. aureus in vivo and low toxicity to mouse tissues. The Ru-8 showed low toxicity to Raw264.7 cells (mouse monocyte macrophage leukemia cells). This study indicates that the ruthenium complex ruthenium quaternary was a promising strategy for the development of new antibacterial agents.

Bacteriophage vB_kpnS-Kpn15: Unveiling its potential triumph against extended-spectrum beta-lactamase-producing Klebsiella pneumoniae - Unraveling efficacy through innovative animal alternate models

Aim –To isolate bacteriophages targeting extended-spectrum beta-lactamase-producing K. pneumoniae and evaluate their effectiveness across diverse models, incorporating innovative alternatives in animal testing. Methods and resultsvB_kpnS-Kpn15 was isolated from sewage sample from Thane district. It produced a clear plaques on K. pneumoniae ATCC 700603. It has a flexible, non-contractile long tail and an icosahedral head and the Siphoviridae family of viruses in the order Caudovirales matched all of its structural criteria. Sequencing of vB_kpnS-Kpn15 revealed a 48,404 bp genome. The vB_KpnS-Kpn15 genome was found to contain 50 hypothetical proteins, of which 16 were found to possess different functions. The vB_KpnS-Kpn15 was also found to possess enzymes for its DNA synthesis. It was found to be lytic for the planktonic cells of K. pneumoniae and bactericidal for up to 48 h and potentially affected established K. pneumoniae biofilms. It demonstrated a broad host range and caused lytic zones on about 46 % of K. pneumoniae multi-drug resistant strains. In an in vitro wound and burn infection model, phage vB_kpnS-Kpn15 in combination with other phages resulted in successful cell proliferation and wound healing. Based on vB_kpnS-Kpn15's lytic properties, it can be incorporated in a bacteriophage cocktail to combat ESBL strains. ConclusionsThe phages isolated during this research are better candidates for phage therapy, and therefore provide new and exciting options for the successful control of antibiotic-resistant bacterial infections in the future. The utilization of animal alternative models in this study elucidates cellular proliferation and migration, underscoring its significance in screening novel drugs with potential applications in the treatment of wound and burn infections. Significance and impact of the researchThe findings of this research have implications for the creation of innovative, promising strategies to treat ESBL K. pneumoniae infections.

Novel benzothiazole derivatives target the Gac/Rsm two-component system as antibacterial synergists against Pseudomonas aeruginosa infections

The management of antibiotic-resistant, bacterial biofilm infections in skin wounds poses an increasingly challenging clinical scenario. Pseudomonas aeruginosa infection is difficult to eradicate because of biofilm formation and antibiotic resistance. In this study, we identified a new benzothiazole derivative compound, SN12 (IC50 = 43.3 nmol/L), demonstrating remarkable biofilm inhibition at nanomolar concentrations in vitro. In further activity assays and mechanistic studies, we formulated an unconventional strategy for combating P. aeruginosa-derived infections by targeting the two-component (Gac/Rsm) system. Furthermore, SN12 slowed the development of ciprofloxacin and tobramycin resistance. By using murine skin wound infection models, we observed that SN12 significantly augmented the antibacterial effects of three widely used antibiotics—tobramycin (100-fold), vancomycin (200-fold), and ciprofloxacin (1000-fold)—compared with single-dose antibiotic treatments for P. aeruginosa infection in vivo. The findings of this study suggest the potential of SN12 as a promising antibacterial synergist, highlighting the effectiveness of targeting the two-component system in treating challenging bacterial biofilm infections in humans.

Bactericidal effectiveness of high-intensity pulsed broadband irradiation in treating infected wounds

The study aimed to investigate the bactericidal eŠcacy of high-intensity pulsed broadband irradiation in the treatment of infected wounds. An experimental study was conducted on 90 mature male Wistar rats. An infected wound model was created by contaminating with Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, and Candida albicans. Animals in Group 1 received high-intensity pulsed broadband irradiation. Animals in Group 2 received traditional UV irradiation. Animals in Group 3 had their wounds cleaned with 0.1% chlorhexidine solution. By the 3rd day of treatment, animals that received pulsed high-intensity broadband irradiation showed a signifiant reduction in contamination by Staphylococcus aureus, Klebsiella pneumoniae, and Pseudomonas aeruginosa compared to Group 3. By the 7th day of treatment, half or the majority of animals in Groups 1 and 2 showed complete decontamination of wounds concerning Staphylococcus aureus and Klebsiella pneumoniae. Most animals in Group 1 showed complete wound clearance of Pseudomonas aeruginosa. By the 10th day, nearly all animals in Group 1 demonstrated complete decontamination of wounds. Statistical analysis revealed a signifiant difference in the reduction of wound contamination with Staphylococcus aureus and Klebsiella pneumoniae by the 10th day in Groups 1 and 2 compared to Group 3. Thus, the use of high-intensity pulsed broadband irradiation of wounds reduces the degree of pathogenic microorganism contamination in a shorter time frame.