Gram-negative Bacterial Biofilms Research Articles

Exploring the dynamics of mixed-species biofilms involving Candida spp. and bacteria in cystic fibrosis.

Cystic fibrosis (CF) is an inherited disease that results from mutations in the gene responsible for the cystic fibrosis transmembrane conductance regulator (CFTR). The airways become clogged with thick, viscous mucus that traps microbes in respiratory tracts, facilitating colonization, inflammation and infection. CF is recognized as a biofilm-associated disease, it is commonly polymicrobial and can develop in biofilms. This review discusses Candida spp. and both Gram-positive and Gram-negative bacterial biofilms that affect the airways and cause pulmonary infections in the CF context, with a particular focus on mixed-species biofilms. In addition, the review explores the intricate interactions between fungal and bacterial species within these biofilms and elucidates the underlying molecular mechanisms that govern their dynamics. Moreover, the review addresses the multifaceted issue of antimicrobial resistance in the context of CF-associated biofilms. By synthesizing current knowledge and research findings, this review aims to provide insights into the pathogenesis of CF-related infections and identify potential therapeutic approaches to manage and combat these complex biofilm-mediated infections.

Biosynthesized silver nanoparticles-capped chondroitin sulfate nanogel targeting microbial infections and biofilms for biomedical applications

Medical devices are essential for patient care, but they can also serve as havens for dangerous microbes and the development of biofilm, which can lead to serious infections and higher death rates. To meet these issues, it is crucial to develop novel and effective antimicrobial coatings for medical devices. In this context, we have developed a new biofunctionalized nanosilver (ICS-Ag), employing itaconyl-chondroitin sulfate nanogel (ICSNG) as a synergistic reducing and stabilizing agent, to effectively eradicate microbial infections and biofilm formation. The antibacterial investigations showed that ICS-Ag nanocomposite is an intriguing antibiotic with excellent antibacterial indices (MIC/MBC (μg/mL): 2.29/4.58, 1.25/2.50, and 1.36/1.36 against S. aureus, E. coli, and P. aeruginosa, respectively), as well as antifungal capacity. Furthermore, ICS-Ag demonstrated efficacy superior to that of the antibiotic (ciprofloxacin, Cipro) against both Gram-positive and Gram-negative bacterial biofilms. TEM images of untreated and treated bacterial strains demonstrate synergistic actions that harm the bacterial cytomembrane, leading to the release of intracellular contents and bacterial death. Interestingly, ICS-Ag shows excellent biocompatibility, with an IC50 value (71.25 μg/mL) higher than MICs against tested microbes. Overall, the ICS-Ag film may provide multifunctional antimicrobial coatings for medical equipment to reduce microbial contamination and biofilm development.

Detection and imaging of bacterial biofilms with glutathione-stabilized gold nanoclusters

Bacterial biofilms colonize chronic wounds and surfaces of medical devices, thus making the development of reliable methods for imaging and detection of biofilms crucial. Although fluorescent identification of bacteria is sensitive and non-destructive, the lack of biofilm-specific fluorescent dyes limits the application of this technique to biofilm detection. Here, we demonstrate, for the first time, that fluorescent glutathione-stabilized gold nanoclusters (GSH-AuNCs) without targeting ligands can specifically interact with extracellular matrix components of Gram-negative and Gram-positive bacterial biofilms resulting in fluorescent staining of bacterial biofilms. By contrast, fluorescent bovine serum albumin-stabilized gold nanoclusters and 11-mercaptoundecanoic acid – stabilized gold nanoclusters do not stain the extracellular matrix of biofilms. According to molecular docking studies, GSH-AuNCs show affinity to several targets in extracellular matrix, including amyloid-anchoring proteins, matrix proteins and polysaccharides. Some experimental evidence was obtained for the interaction of GSH-AuNCs with the lipopolysaccharide (LPS) that was isolated from the matrix of Azospirillum baldaniorum biofilms. Based on GSH-AuNCs properties, we propose a new fluorescent method for the measurement of biofilm biomass with a limit of detection 1.7 × 105 CFU/mL. The sensitivity of the method is 10-fold higher than the standard biofilm quantification with the crystal violet assay. There is a good linear relationship between the fluorescence intensity from the biofilms and the number of CFU from the biofilms in the range from 2.6 × 105 to 6.7 × 107 CFU/mL. The developed nanocluster-mediated method of biofilm staining was successfully applied for quantitative detection of biofilm formation on urinary catheter surface. The presented data suggest that fluorescent GSH-AuNCs can be used to diagnose medical device-associated infections.

Ciprofloxacin-Loaded Mixed Polymeric Micelles as Antibiofilm Agents.

In this work, mixed polymeric micelles (MPMs) based on a cationic poly(2-(dimethylamino)ethyl methacrylate)-b-poly(ε-caprolactone)-b-poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA29-b-PCL70-b-PDMAEMA29) and a non-ionic poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene oxide) (PEO99-b-PPO67-b-PEO99) triblock copolymers, blended at different molar ratios, were developed. The key physicochemical parameters of MPMs, including size, size distribution, and critical micellar concentration (CMC), were evaluated. The resulting MPMs are nanoscopic with a hydrodynamic diameter of around 35 nm, and the ζ-potential and CMC values strongly depend on the MPM's composition. Ciprofloxacin (CF) was solubilized by the micelles via hydrophobic interaction with the micellar core and electrostatic interaction between the polycationic blocks, and the drug localized it, to some extent, in the micellar corona. The effect of a polymer-to-drug mass ratio on the drug-loading content (DLC) and encapsulation efficiency (EE) of MPMs was assessed. MPMs prepared at a polymer-to-drug mass ratio of 10:1 exhibited very high EE and a prolonged release profile. All micellar systems demonstrated their capability to detach pre-formed Gram-positive and Gram-negative bacterial biofilms and significantly reduced their biomass. The metabolic activity of the biofilm was strongly suppressed by the CF-loaded MPMs indicating the successful drug delivery and release. The cytotoxicity of empty and CF-loaded MPMs was evaluated. The test reveals composition-dependent cell viability without cell destruction or morphological signs of cell death.

The role of nanocomposites against biofilm infections in humans.

The use of nanomaterials in several fields of science has undergone a revolution in the last few decades. It has been reported by the National Institutes of Health (NIH) that 65% and 80% of infections are accountable for at least 65% of human bacterial infections. One of their important applications in healthcare is the use of nanoparticles (NPs) to eradicate free-floating bacteria and those that form biofilms. A nanocomposite (NC) is a multiphase stable fabric with one or three dimensions that are much smaller than 100 nm, or systems with nanoscale repeat distances between the unique phases that make up the material. Using NC materials to get rid of germs is a more sophisticated and effective technique to destroy bacterial biofilms. These biofilms are refractory to standard antibiotics, mainly to chronic infections and non-healing wounds. Materials like graphene and chitosan can be utilized to make several forms of NCs, in addition to different metal oxides. The ability of NCs to address the issue of bacterial resistance is its main advantage over antibiotics. This review highlights the synthesis, characterization, and mechanism through which NCs disrupt Gram-positive and Gram-negative bacterial biofilms, and their relative benefits and drawbacks. There is an urgent need to develop materials like NCs with a larger spectrum of action due to the rising prevalence of human bacterial diseases that are multidrug-resistant and form biofilms.

Kinetic Characterization and Computational Modeling of Escherichia coli Heptosyltransferase II: Exploring the Role of Protein Dynamics in Catalysis for GT-B Glycosyltransferase.

Glycosyltransferase (GT) enzymes promote the formation of glycosidic bonds between a sugar molecule and a diversity of substrates. Heptosyltransferase II (HepII) is a GT involved in the lipopolysaccharide (LPS) biosynthetic pathway that transfers the seven-carbon sugar (l-glycero-d-manno-heptose, Hep) onto a lipid-anchored glycopolymer (heptosylated Kdo2-Lipid A, Hep-Kdo2-Lipid A, or HLA). LPS plays a key role in Gram-negative bacterial sepsis, biofilm formation, and host colonization, and as such, LPS biosynthetic enzymes are targets for novel antimicrobial therapeutics. Three heptosyltransferases are involved in the inner-core LPS biosynthesis, with Escherichia coli HepII being the last to be quantitatively characterized in vivo. HepII shares modest sequence similarity with heptosyltransferase I (HepI) while maintaining a high degree of structural homology. Here, we report the first kinetic and biophysical characterization of HepII and demonstrate the properties of HepII that are shared with HepI, including sugar donor promiscuity and sugar acceptor-induced secondary structural changes, which results in significant thermal stabilization. HepII also has an increased catalytic efficiency and a significantly tighter binding affinity for both of its substrates compared to HepI. A structural model of the HepII ternary complex, refined by molecular dynamics simulations, was developed to probe the potentially important substrate-protein contacts. Ligand binding-induced changes in Trp fluorescence in HepII enabled the determination of substrate dissociation constants. Combined, these efforts meaningfully enhance our understanding of the heptosyltransferase family of enzymes and will aid in future efforts to design novel, potent, and specific inhibitors for this family of enzymes.

Facet-Dependent Bactericidal Activity of Ag3PO4 Nanostructures against Gram-Positive/Negative Bacteria

Ag3PO4 nanostructures (APNs) containing silver (Ag metal; of the noble metal families) have the potential to exhibit enzyme-mimetic activity. A nanostructure shape, including its surface facets, can improve the bioactivity of enzyme mimicry, yet the molecular mechanisms remain unclear. Herein, we report facet-dependent peroxidase and oxidase-like activity of APNs with both antibacterial and biofilm degrading properties through the generation of reactive oxygen species. Cubic APNs had superior antibacterial effects than rhombic dodecahedral shapes when inhibiting Gram-positive and Gram-negative bacterial pathogen proliferation and biofilm degradation. A similar performance was observed for rhombic dodecahedral shapes, being greater than tetrahedral-shaped APNs. The extent of enzyme-mimetic activity is attributed to the facets {100} present in cubic APNs that led the peroxide radicals to inhibit the proliferation of bacteria and degrade biofilm. These facets were compared to rhombic dodecahedral APNs {110} and tetrahedral APNs {111}, respectively, to reveal a facet-dependent enhanced antibacterial activity, providing a plausible mechanism for shape-dependent APNs material enzyme-mimetic effects on bacteria. Thus, our research findings can provide a direction to optimize bactericidal materials using APNs in clinically relevant applications.

A novel smaller β-defensin-derived peptide is active against multidrug-resistant bacterial strains.

Antibiotic resistance is becoming a severe obstacle in the fight against acute and chronic infectious diseases that accompany most degenerative illnesses from neoplasia to osteo-arthritis and obesity. Currently, the race is on to identify pharmaceutical molecules or combinations of molecules able to prevent or reduce the insurgence and/or progression of infectivity. Attempts to substitute antibiotics with antimicrobial peptides have, thus far, met with little success against multidrug-resistant (MDR) bacterial strains. During the last decade, we designed and studied the activity and features of human β-defensin analogs, which are salt-resistant, and hence active also under high salt concentrations as, for instance, in cystic fibrosis. Herein, we describe the design, synthesis, and major features of a new 21 aa long molecule, peptide γ2. The latter derives from the γ-core of the β-defensin natural molecules, a small fragment of these molecules still bearing high antibacterial activity. We found that peptide γ2, which contains only one disulphide bond, recapitulates most of the biological properties of natural human β-defensins and can also counteract both Gram-positive and Gram-negative MDR bacterial strains and biofilm formation. Moreover, it has great stability in human serum thereby enhancing its antibacterial presence and activity without cytotoxicity in human cells. In conclusion, peptide γ2 is a promising new weapon also in the battle against intractable infectious diseases.

Functional Nanomaterials and 3D-Printable Nanocomposite Hydrogels for Enhanced Cell Proliferation and for the Reduction of Bacterial Biofilm Formation.

Biomaterial-associated infections are a major cause of biomaterial implant failure. To prevent the initial attachment of bacteria to the implant surface, researchers have investigated various surface modification methods. However, most of these approaches also prevent the attachment, spread, and growth of mammalian cells, resulting in tissue integration failure. Therefore, the success of biomaterial implants requires an optimal balance between tissue integration (cell adhesion to biomaterial implants) and inhibition of bacterial colonization. In this regard, we synthesize bifunctional nanomaterials by functionalizing the pores and outer surfaces of periodic mesoporous organosilica (PMO) with antibacterial tetracycline (Tet) and antibacterial and cell-adhesive bipolymer poly-d-lysine (PDL), respectively. Then, the fabricated TetPMO-PDL nanomaterials are incorporated into alginate-based hydrogels to create injectable and 3D-printable nanocomposite (NC) hydrogels (AlgL-TetPMO-PDL). These bifunctional nanomaterial and 3D-printable NC hydrogel show pH-dependent release of Tet over 7 days. They also enhance the proliferation of eukaryotic cells (fibroblasts). TetPMO-PDL is inactive in reducing Pseudomonas aeruginosa, Staphylococcus aureus, and Enterococcus faecalis biofilms. However, AlgL-TetPMO-PDL shows significant antibiofilm activity against P. aeruginosa. These results suggest that the incorporation of TetPMO-PDL into AlgL may have a synergistic effect on the inhibition of the Gram-negative bacterial (P. aeruginosa) biofilm, while this has no effect on the reduction of the Gram-positive bacterial (S. aureus and E. faecalis) biofilm.

Bacterial metal nanoparticles to develop new weapons against bacterial biofilms and infections.

The widespread use of antibiotics has resulted in the outbreak and spread of antibiotic-resistant pathogens. Bacterial antibiotic resistance may develop at cellular and community levels. In the latter case, it is based on tolerance which implicates the shift from a free-living form of life (i.e., planktonic) to a sessile multi-stratified community (i.e., biofilm). Metal nanoparticles (MNPs) have been shown to be promising candidates as antimicrobial agents. MNPs are able to interact with and penetrate bacterial biofilms, thus, resulting effective antibiofilm compounds. Another interesting aspect is the possibility of using plants, fungi, yeasts, and bacteria to obtain biogenic MNPs (BMNP). Bacteria are able to grow in presence of many different toxic heavy metal ions thanks to different metal resistance gene clusters that allow a variety of biochemical counters (formation of harmless complexes, efflux, precipitation, reduction, etc.). The formation of BMNPs by bacterial cells could be, in most cases, just a consequence of metal detoxification mechanisms. This review focuses on BMNPs from bacterial origin that may represent a good source of compounds with a broad spectrum of activity against common Gram-positive and Gram-negative pathogens and bacterial biofilms thereof. In particular, the state of art on BMNP synthesis by bacteria is presented and potential applications in the fight against biofilm-associated infections and resistant pathogens are highlighted. In addition, critical aspects on BMNP bacterial synthesis and utilization are commented.Key points• New antimicrobials to fight antibiotic-resistant pathogens are urgently needed.• Biogenic metal nanoparticles can efficiently hit biofilm-forming pathogens.• Metal-nanoparticle composition could confer specific antibiofilm activity.

Antibiofilm activity of Fmoc-phenylalanine against Gram-positive and Gram-negative bacterial biofilms.

Biofilm associated infections are the major contributor of mortality, morbidity and financial burden in patients with a bacterial infection. About 65% of all bacterial infections are associated with the information of bacterial biofilms. Bacterial biofilms not only reduce the efficacy of antibacterial treatment but also increases the threat of developing antibacterial resistance. Recently, our group has discovered the antibacterial activity of Fmoc-phenylalanine (Fmoc-F) and other Fmoc-amino acids (Fmoc-AA). Fmoc-F and other Fmoc-AA showed antibacterial activity due to their surfactant properties. Surfactants are known to eradicate biofilm and enhance antimicrobial activity in biofilm. Thus, in the present study, we evaluated the anti-biofilm activity of Fmoc-F against clinically relevant bacteria. We found that Fmoc-F not only inhibits the biofilm formation in Staphylococcus aureus and Pseudomonas aeruginosa, but also eradicates the already formed biofilms over the surface. Further, Fmoc-F coated glass surface resists S. aureus and P. aeruginosa biofilm formation and attachment, when biofilm is grown over the surface. The mechanistic investigation suggests that Fmoc-F reduces the extracellular matrix (ECM) components such as proteins, carbohydrates and eDNA in the biofilm and affect its stability via direct interactions with ECM components and/ or indirectly through reducing bacterial cell population. Finally, we showed that Fmoc-F treatment in combination with vancomycin and ampicillin synergistically inhibit biofilm formation. Overall, the study demonstrates the potential application of Fmoc-F and other Fmoc-AA molecules individually as well as in combination as anti-biofilm coating material for treating biofilm associated infections.

In Vitro Destruction of Pathogenic Bacterial Biofilms by Bactericidal Metallic Nanoparticles via Laser-Induced Forward Transfer.

A novel, successful method of bactericidal treatment of pathogenic bacterial biofilms in vitro by laser-induced forward transfer of metallic nanoparticles from a polyethylene terephthalate polymeric substrate was suggested. Transferred nanoparticles were characterized by scanning and transmission electron microscopy, energy-dispersive X-ray and Raman spectroscopy. The antibacterial modality of the method was tested on Gram-positive (Staphylococcus aureus) and Gram-negative (Pseudomonas Aeruginosa) bacterial biofilms in vitro, revealing their complete destruction. The proposed simple, cost-effective and potentially mobile biofilm treatment method demonstrated its high and broad bactericidal efficiency.

Impact of the antibiotic-cargo from MSNs on Gram-positive and Gram-negative bacterial biofilms.

Mesoporous silica nanoparticles (MSNs) are promising drug nanocarriers for infection treatment. Many investigations have focused on evaluating the capacity of MSNs to encapsulate antibiotics and release them in a controlled fashion. However, little attention has been paid to determine the antibiotic doses released from these nanosystems that are effective against biofilm during the entire release time. Herein, we report a systematic and quantitative study of the direct effect of the antibiotic-cargo released from MSNs on Gram-positive and Gram-negative bacterial biofilms. Levofloxacin (LVX), gentamicin (GM) and rifampin (RIF) were separately loaded into pure-silica and amino-modified MSNs. This accounts for the versatility of these nanosystems since they were able to load and release different antibiotic molecules of diverse chemical nature. Biological activity curves of the released antibiotic were determined for both bacterial strains, which allowed to calculate the active doses that are effective against bacterial biofilms. Furthermore, in vitro biocompatibility assays on osteoblast-like cells were carried out at different periods of times. Albeit a slight decrease in cell viability was observed at the very initial stage, due to the initial burst antibiotic release, the biocompatibility of these nanosystems is evidenced since a recovery of cell viability was achieved after 72 h of assay. Biological activity curves for GM released from MSNs exhibited sustained patterns and antibiotic doses in the 2-6 μg/mL range up to 100 h, which were not enough to eradicate biofilm. In the case of LVX and RIF first-order kinetics featuring an initial burst effect followed by a sustained release above the MIC up to 96 h were observed. Such doses reduced by 99.9% bacterial biofilm and remained active up to 72 h with no emergence of bacterial resistance. This pioneering research opens up promising expectations in the design of personalized MSNs-based nanotherapies to treat chronic bone infection.

Improved hemocompatibility and reduced bacterial adhesion on superhydrophobic titania nanoflower surfaces

Thrombosis formation and bacterial infection are key challenges for blood-contacting medical devices. When blood components encounter a device's surface, proteins are adsorbed, followed by the adhesion and activation of platelets as well as an immune response. This culminates in clot formation via the trapping of red blood cells in a fibrin matrix, which can block the device's function and cause severe complications for the patient. In addition, bacteria may adhere to a device's surface. This can lead to the formation of a biofilm, a protective layer for bacteria that significantly increases resistance to antibiotics. Despite years of research, no long-term solutions have been discovered to combat these issues. To impede thrombosis, patients often take antiplatelet drugs for the life of their device, which can cause excess bleeding and other complications. Patients can take antibiotics to fight bacterial infection, but these are often ineffective if biofilms are formed. Superhydrophobic surfaces show promise in reducing both thrombosis and bacterial infection on devices by impeding contact between biological components and the biomaterial. In this study, superhydrophobic titania nanoflower surfaces were successfully fabricated on a titanium alloy Ti-6Al-4V substrate with hydrothermal synthesis and vapor-phase silanization. The surface topography, surface wettability, surface chemistry, and surface crystallography of the surfaces was subsequently characterized. Surface hemocompatibility was investigated through lactate dehydrogenase (LDH) cytotoxicity analysis, blood-plasma protein adsorption, platelet and leukocyte adhesion and activation, and whole blood clotting analysis. Surface bacterial infection was characterized through Gram-positive and Gram-negative bacterial adhesion and biofilm morphology. The results indicated a reduction of protein adsorption, platelet and leukocyte adhesion and activation, bacterial adhesion, and biofilm formation as well as improved contact angle stability compared to control surfaces.

Synergy between Sophorolipid Biosurfactant and SDS Increases the Efficiency of P. aeruginosa Biofilm Disruption

Biofilms are communities of bacteria encased in self-secreted extracellular polymeric substances (EPS) that adhere stubbornly to submerged surfaces. Once established, these communities can cause serious chronic illnesses in medical settings, while they can promote corrosion and biofouling in industrial settings. Due to the difficulty of their removal, strongly oxidizing chemicals and detergents can be used to degrade and remove biofilms by killing the cells and degrading the matrix; however, the choice of compounds is limited in delicate environments due to the potential damage they may cause. In the case of detergents, most are synthesized from nonrenewable petrochemicals that have a degree of aquatic toxicity. There is a growing need to identify and characterize alternatives to synthetic surfactants. Biosurfactants, which are surfactants produced by microorganisms, are a promising alternative since they can be synthesized from renewable resources, have low environmental toxicity, and have been shown to have higher degrees of specificity in the mechanism of action. Sophorolipids are a class of glycolipid surfactants produced by yeast that have demonstrated great promise due to large yields from renewable feedstocks and for antimicrobial properties; however, the effect of the application of sophorolipids to Gram-negative bacterial biofilms has not been well studied. We investigate the antibiofilm properties of sophorolipids by demonstrating its ability to cause the catastrophic disruption of Pseudomonas aeruginosa PAO1 biofilms in microfluidic channels. We show that while sophorolipids inflict little damage to the bacteria, they weaken the EPS biofilm matrix, leading to surface-detachment and breakup of the biofilm. Furthermore, we find that sophorolipids act cooperatively with the widely used surfactant, sodium dodecyl sulfate. When combined, concentrations ∼100-fold lower than the minimum effective concentration, when used independently, recover potency. Biosurfactants are typically expensive to produce, thus our work demonstrates a means to improve efficacy while simultaneously reducing both cost and the amount of environmentally harmful substances used.

Functionalized Polymers Enhance Permeability of Antibiotics in Gram-negative MDR Bacteria and Biofilms for Synergistic Antimicrobial Therapy.

The emergence of multi-drug resistant pathogenic bacteria constitutes a key threat to global health. Infections caused by multi-drug resistant Gram-negative bacteria are particularly challenging to treat due to the ability of pathogens to prevent antibiotic penetration inside the bacterial membrane. Antibiotic therapy is further rendered ineffective due to biofilm formation where the protective Extracellular Polymeric Substance (EPS) matrix limits the diffusion of antibiotics inside the biofilm. We hypothesized that careful engineering of chemical groups on polymer scaffolds could enable polymers to penetrate the barriers of Gram-negative bacterial membrane and biofilm matrix. Here, we present the use of engineered polymeric nanoparticles in combination with antibiotics for synergistic antimicrobial therapy. These polymeric nanoparticles enhance the accumulation of antibiotics inside Gram-negative bacteria and biofilm matrix, resulting in increased potency of antibiotics in combination therapy. Sub-lethal concentrations of engineered polymeric nanoparticles reduce the antibiotic dosage by 32-fold to treat MDR bacteria and biofilms. Tailoring of chemical groups on polymers demonstrate a strong-structure activity relationship in generating additive and synergistic combinations with antibiotics. This study demonstrates the ability of polymeric nanoparticles to 'rejuvenate' antibiotics rendered ineffective by resistant bacteria and provides a rationale to design novel compounds to achieve effective antimicrobial combination therapies.

Identification of new dermaseptins with self-assembly tendency: membrane disruption, biofilm eradication, and infected wound healing efficacy

Severe infections associated with antibiotic-resistant bacteria and biofilms have attracted increasing interest as these diseases are difficult to treat with current antibiotics. Typical cationic antimicrobial peptides dermaseptins are considered to be the most promising next-generation antibiotics because of their broad-spectrum antimicrobial activities and minor side effects. Two new dermaseptin peptides, DMS-PS1 and DMS-PS2, have been identified by “shotgun” molecular cloning of encoding cDNAs in the crude skin secretions of the waxy monkey tree frog, Phyllomedusa sauvagei. The mature peptide sequences predicted from the cloned cDNAs were separated from crude skin secretions and confirmed by mass spectrometry. Chemically synthetic replicates were assessed for various biological activities. Both dermaseptins were potently effective against a broad spectrum of microorganisms including antibiotic-resistant bacteria and displayed significant potency against gram-positive and gram-negative bacterial biofilms with low toxicity towards mammalian red blood cells. Remarkably, DMS-PS2 was effective against infections in murine skin caused by methicillin-resistant Staphylococcus aureus as a result of an induced wound. The actions of DMS-PS2 were with a membrane permeabilization mode. Overall, the data provided convincing evidence for the development of anti-infectious agents and/or biomaterials as a new therapeutic approach against bacterial infections. Statement of SignificanceBacterial adhesion to biomaterials remains a major problem. Antimicrobial peptides (AMPs) are well-known components of the innate immune system that can be applied to overcome biofilm-associated infections. Cationic dermaseptin peptides showed significant broad-spectrum antimicrobial activities and activities against bacterial biofilms of persistent infections in association with weak toxicity for mammalian red blood cells. The membrane permeabilizing ability of DMS-PS2 was confirmed, and importantly, it demonstrated potent efficiency of the treatment of MRSA infected murine skin model. Furthermore, beyond our expectation, DMS-PS2 showed a self-aggregating parameter, indicating a promising potential for the use of immobilized AMPs in clinical applications., which makes it also a promising suggestion for infection-proof biomaterial development.

Antibacterial Liquid Metals: Biofilm Treatment via Magnetic Activation.

Antibiotic resistance has made the treatment of biofilm-related infections challenging. As such, the quest for next-generation antimicrobial technologies must focus on targeted therapies to which pathogenic bacteria cannot develop resistance. Stimuli-responsive therapies represent an alternative technological focus due to their capability of delivering targeted treatment. This study provides a proof-of-concept investigation into the use of magneto-responsive gallium-based liquid metal (LM) droplets as antibacterial materials, which can physically damage, disintegrate, and kill pathogens within a mature biofilm. Once exposed to a low-intensity rotating magnetic field, the LM droplets become physically actuated and transform their shape, developing sharp edges. When placed in contact with a bacterial biofilm, the movement of the particles resulting from the magnetic field, coupled with the presence of nanosharp edges, physically ruptures the bacterial cells and the dense biofilm matrix is broken down. The antibacterial efficacy of the magnetically activated LM particles was assessed against both Gram-positive and Gram-negative bacterial biofilms. After 90 min over 99% of both bacterial species became nonviable, and the destruction of the biofilms was observed. These results will impact the design of next-generation, LM-based biofilm treatments.

Photodynamic inactivation of Pseudomonas aeruginosa bacterial biofilms using photosensitizers based on octacationic derivatives of phthalocyanines and bacteriochlorins

Antibacterial photodynamic therapy is a promising method of treating local infected foci, especially surgical and burn wounds, trophic and diabetic ulcers. This work explores the photophysical and antibacterial properties of novel phthalocyanine- and synthetic-bacteriochlorin-based octacationic photosensitizers (PS). The results of the study confirm their low degree of aggregation at high concentrations, as well as high efficiency of photodynamic treatment of Gram-negative bacterial biofilms.

Toward the Development of an Innovative Implant: NiTi Alloy Functionalized by Multifunctional β-TCP+Ag/SiO2 Coatings.

In recent years, one of the more important and costly problems of modern medicine is the need to replace or supplement organs in order to improve the quality of human life. In this field, promising solutions seem to have been implants which are based on NiTi alloys with shape memory effects. Unfortunately, this material is susceptible to the corrosion and release of toxic nickel to the human organism. Hence, its application as a long-term material is strongly limited. Therefore, this paper presents a new solution which should help to improve the functionality of the NiTi alloy and elongate its medical stability to use. The idea was focused on functionalization of the implant surface by a biocompatible, multifunctional coating without any impact on the features of the substrate, i.e., the martensitic transformation responsible for shape memory effects. For this purpose, we prepared a colloidal suspension, composed of β-TCP (particle size ∼450 nm) and the Ag/SiO2 nanocomposite which due to the electrophoretic deposition (EPD) led to the formation of structurally atypical calcium phosphosilicate coating. Those biomaterials formed a crack-free coating, adhering well to the NiTi surface when distributed over the entire surface, with low concentration of metallic and oxide silver (<3 at. %). At the same time, the coat-forming materials had resulted in the growth of a Gram-negative bacterial biofilm. Additionally, the additive of the silver-silica composite enhances cell proliferation, effectively a few times higher than commonly used coat-forming materials (e.g., pure β-TCP).