Killing Of Methicillin-resistant Staphylococcus Aureus Research Articles

Gelatin emulsion gels loaded with host defence peptides for the treatment of antibiotic-resistant infections

The surge in multidrug-resistant bacteria against conventional antibiotics is a rapidly developing global health crisis necessitating novel infection management strategies. Host defence peptides (HDPs), also known as antimicrobial peptides (AMPs), offer a promising alternative to traditional antibiotics, but their practical translation is limited by their susceptibility to proteases and potential off-site cytotoxicity. In this paper, we investigate the feasibility of using gelatin emulsion gels (GELs), prepared using a water-in-oil (W/O) method, for the delivery of HDPs DJK-5 and IDR-1018 to improve their clinical utility. DJK-5-loaded GELs exhibited complete eradication of planktonic Methicillin-resistant Staphylococcus aureus (MRSA) at 4 - and 24-h intervals. Similarly, IDR-1018-loaded GELs demonstrated almost complete killing of MRSA and Escherichia coli (E. coli) after 4 h. Importantly, none of the GEL formulations investigated exhibited in vitro cytotoxicity. Overall, these HDP loaded GELs are a promising solution for the treatment of antibiotic-resistant infections.

Riboflavin-Vancomycin Conjugate Enables Simultaneous Antibiotic Photo-Release and Photodynamic Killing against Resistant Gram-Positive Pathogens.

Decades of antibiotic misuse have led to alarming levels of antimicrobial resistance, and the development of alternative diagnostic and therapeutic strategies to delineate and treat infections is a global priority. In particular, the nosocomial, multidrug-resistant "ESKAPE" pathogens such as Gram-positive methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus spp (VRE) urgently require alternative treatments. Here, we developed light-activated molecules based on the conjugation of the FDA-approved photosensitizer riboflavin to the Gram-positive specific ligand vancomycin to enable targeted antimicrobial photodynamic therapy. The riboflavin-vancomycin conjugate proved to be a potent and versatile antibacterial agent, enabling the rapid, light-mediated, killing of MRSA and VRE with no significant off-target effects. The attachment of riboflavin on vancomycin also led to an increase in antibiotic activity against S. aureus and VRE. Simultaneously, we evidenced for the first time that the flavin subunit undergoes an efficient photoinduced bond cleavage reaction to release vancomycin, thereby acting as a photoremovable protecting group with potential applications in drug delivery.

Probiotic disruption of quorum sensing reduces virulence and increases cefoxitin sensitivity in methicillin-resistant Staphylococcus aureus

Therapies which target quorum sensing (QS) systems that regulate virulence in methicillin-resistant Staphylococcus aureus (MRSA) are a promising alternative to antibiotics. QS systems play a crucial in the regulation of MRSA antibiotic resistance, exotoxin production, antioxidant protection and immune cell evasion, and are therefore attractive therapeutic targets to reduce the virulence of a pathogen. In the present work the the effects of bioactive peptides isolated from two strains of lactic acid bacteria were tested against antibiotic resistance, carotenoid production, resistance to oxidative killing and biofilm structure in two clinical MRSA isolates. The results obtained from fractional-inhibitory concentration assays with bulk and semi-purified bioactive molecules showed a significant synergistic effect increasing cefoxitin mediated killing of MRSA. This was coupled to a six-fold decrease of the major membrane pigment staphyloxanthin, and a 99% increase in susceptibility to oxidative stress mediated killing. Real-time quantitative PCR analysis of the QS-genes agrA and luxS, showed differential expression between MRSA strains, and a significant downregulation of the hemolysin gene hla. Light microscopy and scanning electron microscopy revealed alteration in biofilm formation and clustering behavior. These results demonstrate that bioactive metabolites may be effectively applied in tandem with beta-lactam antibiotics to sensitize MRSA to cefoxitin. Moreover, these results shown that several key QS-controlled virulence mechanisms are diminished by probiotic metabolites.

Soluble C-Type Lectin-Receptor Ligands Stimulate ROS Production in Dendritic Cells and Potentiate Killing of MRSA as Well as the MRSA Induced IL-12 Production.

Methicillin resistant Staphylococcus aureus (MRSA) has developed resistance to most β-lactam antibiotics leaving few treatment options against infections with MRSA. Through mannose receptors, mannan potentiates IL-12 production induced by Gram-positive bacteria, a cytokine crucial in the clearance of S. aureus infection. We investigated the IL-12 potentiating effect of mannan pre-treatment of bone marrow-derived dendritic cells prior to stimulation with clinical MRSA strains. Mannan almost doubled IL-12 as well as IFN-β production in response to USA300, also when USA300 was treated with the β-lactam cefoxitin. The MRSA-induced IL-12 production was dependent on bacterial uptake and reactive oxygen species (ROS). Mannan alone induced ROS production, and in combination with USA300, the ROS produced corresponded to the sum induced by mannan and USA300. Addition of a monoclonal antibody against the mannose receptor likewise enhanced USA300-induced IL-12 and induced ROS production. Mannan addition further increased the endocytosis as well as the rate of endosomal killing of bacteria. Pre-treatment with soluble β-glucans also induced ROS and potentiated the USA300-induced IL-12 indicating that other C-type receptors may play a similar role. In the presence of the pro-inflammatory mediators, GM-CSF or IFN-γ, the mannan-enhanced IL-12 production increased further. The USA300-induced and the mannan-facilitated enhanced IFN-β and IL-12 showed same dependency on MAPK c-Jun N-terminal kinase signaling, suggesting that mannan enhances the signals already induced by the bacteria, rather than changing them. We suggest that the C-type lectin-induced ROS production is a key factor in the IFN-β and IL-12 potentiation.

Fibrotic lung disease inhibits immune responses to staphylococcal pneumonia via impaired neutrophil and macrophage function.

Idiopathic pulmonary fibrosis (IPF) is a progressive and fatal disease characterized by collagen deposition within the lung interstitium. Bacterial infection is associated with increased morbidity and more rapid mortality in IPF patient populations, and pathogens such as methicillin-resistant Staphylococcus aureus (MRSA) are commonly isolated from the lungs of hospitalized patients with IPF. Despite this, the effects of fibrotic lung injury on critical immune responses to infection remain unknown. In the present study, we show that, like humans with IPF, fibrotic mice infected with MRSA exhibit increased morbidity and mortality compared with uninfected fibrotic mice. We determine that fibrosis conferred a defect in MRSA clearance compared with nonfibrotic mice, resulting from blunted innate immune responses. We show that fibrosis inhibited neutrophil intracellular killing of MRSA through impaired neutrophil elastase release and oxidative radical production. Additionally, we demonstrate that lung macrophages from fibrotic mice have impaired phagocytosis of MRSA. Our study describes potentially novel impairments of antimicrobial responses upon pulmonary fibrosis development, and our findings suggest a possible mechanism for why patients with IPF are at greater risk of morbidity and mortality related to infection.

Inhibition of Bacterial Efflux Pumps by Crude Extracts and Essential Oil from Myristica fragrans Houtt. (Nutmeg) Seeds against Methicillin-Resistant Staphylococcus aureus.

Myristicafragrans Houtt. (Nutmeg) is a widely known folk medicine across several parts of Asia, particularly used in antimicrobial treatment. Bacterial resistance involves the expression of efflux pump systems (chromosomal norA and mepA) in methicillin-resistant Staphylococcus aureus (MRSA). Crude extract (CE) and essential oil (EO) obtained from nutmeg were applied as efflux pump inhibitors (EPIs), thereby enhancing the antimicrobial activity of the drugs they were used in. The major substances in CE and EO, which function as EPIs, in a descending order of % peak area include elemicin, myristicin, methoxyeugenol, myristicin, and asarone. Here, we investigated whether the low amount of CE and EO used as EPIs was sufficient to sensitize MRSA killing using the antibiotic ciprofloxacin, which acts as an efflux system. Interestingly, synergy between ciprofloxacin and CE or EO revealed the most significant viability of MRSA, depending on norA and mepA, the latter being responsible for EPI function of EO. Therefore, CE and EO obtained from nutmeg can act as EPIs in combination with substances that act as efflux systems, thereby ensuring that the MRSA strain is susceptible to antibiotic treatment.

DNA Aptamer-Conjugated Magnetic Graphene Oxide for Pathogenic Bacteria Aggregation: Selective and Enhanced Photothermal Therapy for Effective and Rapid Killing.

Methicillin-resistant Staphylococcus aureus (MRSA), often called “superbug”, is a nosocomial and multidrug resistance bacterium that shows resistance to β-lactam antibiotics. There has been high demand to develop an alternative treatment model to antibiotics for efficiently fighting MRSA. Herein, we developed DNA aptamer-conjugated magnetic graphene oxide (Apt@MGO) as a multifunctional and biocompatible nanoplatform for selective and rapid eradication of MRSA and evaluated heat generation and cell death performance of Apt@MGO for the first time under dispersed and aggregated states. The aptamer sequence was specifically selected for MRSA and acted as a molecular targeting probe for selective MRSA recognition and antibiotic-free therapy. Magnetic graphene oxide (MGO) serves as a nanoplatform for aptamer conjugation and as a photothermal agent by converting near-infrared (NIR) light to heat. Iron oxide nanoparticles (Fe3O4 NPs) are formed on GO to prepare MGO, which shows magnetic properties for collecting MRSA cells in a certain area in the reaction tube by an external magnet. The collected MGO induces remarkably high local heating and eventual MRSA cell death under NIR laser irradiation. We demonstrate that Apt@MGO resulted in ∼78% MRSA and over >97% MRSA cell inactivation in dispersed and aggregated states, respectively, under 200 seconds (sn) exposure of NIR irradiation (808 nm, 1.1 W cm–2). An in vitro study highlights that Apt@MGO is considered a targeted, biocompatible, and light-activated photothermal agent for efficient and rapid killing of MRSA in the aggregated state under NIR light.

Contrasting effects of linezolid on healthy and dysfunctional human neutrophils: reducing C5a-induced injury

Methicillin-resistant Staphylococcus aureus (MRSA) is an important cause of ventilator-associated pneumonia (VAP). Patients with VAP have poorly functioning neutrophils, related to increased levels of the complement fragment C5a. The antibiotic linezolid has been useful in controlling MRSA-related VAP infections; however clinical benefit does not always correlate with antimicrobial effect, suggesting the possibility of immunomodulatory properties. Here the effects of linezolid on healthy and dysfunctional neutrophils (modelled by C5a-induced injury) was investigated. Functional assays (killing, phagocytosis, transmigration, and respiratory burst) were used to assess the effects of pre-, co- and post-incubating linezolid (0.4–40 mg/L) with healthy neutrophils relative to those with C5a-induced injury. C5a decreased neutrophil killing, and phagocytosis of MRSA. Furthermore, C5a significantly decreased neutrophil transmigration to IL-8, but did not affect respiratory burst. Co-incubation of linezolid significantly improved killing of MRSA by dysfunctional neutrophils, which was supported by concomitant increases in phagocytosis. Conversely linezolid impaired killing responses in healthy neutrophils. Pre- or post-incubation of linezolid prior or following C5a induced injury had no effect on neutrophil function. This study suggests that linezolid has immunomodulatory properties that protect human neutrophils from injury and provides insight into its mode of action beyond a basic antibiotic.

Dual-wavelength photo-killing of methicillin-resistant Staphylococcus aureus.

With the effectiveness of antimicrobials declining as antimicrobial resistance continues to threaten public health, we must look to alternative strategies for the treatment of infections. In this study, we investigated an innovative, drug-free, dual-wavelength irradiation approach that combines 2 wavelengths of light, 460 nm and 405 nm, against methicillin-resistant Staphylococcus aureus (MRSA). MRSA was initially irradiated with 460-nm light (90-360 J/cm2) and subsequently irradiated with aliquots of 405-nm light (54-324 J/cm2). For in vivo studies, mouse skin was abraded and infected with approximately 107 CFUs of MRSA and incubated for 3 hours before irradiating with 460 nm (360 J/cm2) and 405 nm (342 J/cm2). Naive mouse skin was also irradiated to investigate apoptosis. We found that staphyloxanthin, the carotenoid pigment in MRSA cells, promoted resistance to the antimicrobial effects of 405-nm light. In addition, we found that the photolytic effect of 460-nm light on staphyloxanthin attenuated resistance of MRSA to 405-nm light killing. Irradiation of 460 nm alone did not elicit any antimicrobial effect on MRSA. In a proof-of-principle mouse skin abrasion infection model, we observed significant killing of MRSA using the dual-wavelength irradiation approach. However, when either wavelength of light was administered alone, no significant decrease in bacterial viability was observed. Moreover, exposure of the dual-wavelength irradiation to naive mouse skin did not result in any visible apoptosis. In conclusion, a dual-wavelength irradiation strategy may offer an innovative, effective, and safe approach for the treatment of skin infections caused by MRSA.

Cardiolipin dynamics regulate proinflammatory cytokine production in methicillin-resistant Staphylococcus aureus--infected macrophages

Abstract During the immune response, mitochondria serve as a hub for innate immune signaling molecules, such as mitochondrial antiviral signaling protein (MAVS) and Nod Like Receptor Family Pyrin Domain Containing 3 (NLRP3). Cardiolipin, a class of phospholipids found primarily in the inner mitochondrial membrane (IMM), regulates mitochondrial function and cellular stress. In lipopolysaccharide (LPS)-stimulated macrophages, CL translocates from the IMM to the outer mitochondrial membrane (OMM) and recruits NLRP3 to promote inflammasome activation. However, the role of CL in macrophage antibacterial responses remains ill defined. Here, we have made CL-deficient RAW264.7 (RAW) cells by stably knocking down Cardiolipin Synthase (Crls1). Upon infection with methicillin-resistant Staphylococcus aureus (MRSA), Crls1 KD RAW cells showed markedly decreased expression and production of cytokines IL-6, IL-1β, and IFN-β compared to control cells, while TNF-α levels remained similar. For IL-6 and IFN-β, transcript and protein levels in Crls1 KD RAW cells were comparable to unstimulated controls. Surprisingly, these cells showed normal MRSA killing and NF-kB p65 nuclear translocation. These findings suggest a novel role for CL in transcript-level regulation of cytokines during infection. To further investigate CL-dependent innate immune functions, we will use RAW cells expressing a GFP-tagged CL-binding protein, Stomatin-like Protein-2-GFP, to track CL dynamics at membrane-level resolution using 3D structured illumination microscopy. This approach supports earlier studies that describe increased CL OMM localization in response to LPS. Our data suggest an underappreciated role for CL in macrophage responses to MRSA infection.

The Droplet-Size Effect Of Squalene@cetylpyridinium Chloride Nanoemulsions On Antimicrobial Potency Against Planktonic And Biofilm MRSA.

BackgroundIt is important to explore the interaction between antibacterial nanoparticles and microbes for understanding bactericidal activity and developing novel applications. It is possible that the nanoparticulate size can govern the antibacterial potency.PurposeThe purpose of this study was to evaluate the antimicrobial and antibiofilm properties of cetylpyridinium chloride (CPC)–decorated nanoemulsions against methicillin-resistant Staphylococcus aureus (MRSA).MethodsThe droplet size could be adjusted by varying the percentage of squalene, the main ingredient of the oily core.ResultsWe fabricated cationic nanoemulsions of three different sizes, 55, 165, and 245 nm. The nanoemulsions showed greater storage stability than the self-assembled CPC micelles. The tested nanoemulsions exhibited more antimicrobial activity against Gram-positive bacteria than Gram-negative bacteria and fungi. The killing of MRSA was mainly induced by direct cell-membrane damage. This rupture led to the leakage of cytoplasmic DNA and proteins. The nanoemulsions might also degrade the DNA helix and disturb protein synthesis. The proteomic analysis indicated the significant downregulation of DNA-directed RNA polymerase (RNAP) subunits β and β’. The antibacterial effect of nanoemulsions increased with decreasing droplet size in the biofilm MRSA but not planktonic MRSA. The small-sized nanoemulsions had potent antibiofilm activity that showed a colony-forming unit (CFU) reduction of 10-fold compared with the control. The loss of total DNA concentration also negatively correlated with the nanoemulsion size.ConclusionThe present report established a foundation for the development of squalene@CPC nanosystems against drug-resistant S. aureus.

Photothermal inactivation of methicillin-resistant Staphylococcus aureus: anti-biofilm mediated by a polypyrrole-carbon nanocomposite.

Widespread resistance to antibiotics amongst pathogens has become a tremendous challenge of high morbidity and mortality rates which increases the needs to exploring novel methods of treatment. An efficient antimicrobial procedure to root out pathogenic bacteria is photothermal therapy. In this study, antimicrobial effects of a polypyrrole-carbon nanocomposite (PPy-C) upon laser irradiation in order to destroy the pathogenic gram-positive bacterium, methicillin-resistant Staphylococcus aureus (MRSA) were assessed. The bacterial cells were incubated with 500, 750 and 1000 μg ml-1 concentrations of PPy-C and irradiated with an 808-nm laser at a power density of 1.0 W cm-2. To indicate the biocompatibility and toxic effect of the nanocomposite without and with laser irradiation, the authors counted the number of CFUs and compared it to an untreated sample. Antibacterial mechanisms of PPy-C were assessed through temperature increment, reactive oxygen species production, and protein and DNA leakages. Photothermal heating assay showed that 26°C temperature increases in the presence of 1000 µg ml-1 PPy-C led to >98% killing of MRSA. Furthermore, 20 min radiation of near-infrared light to PPy-C in different concentrations indicated destruction and reduction in the MRSA biofilm formation. Therefore, PPy-C was introduced as a photothermal absorber with a bactericidal effect in MRSA.

IL-21/type I interferon interplay regulates neutrophil-dependent innate immune responses to Staphylococcus aureus.

Methicillin-resistant Staphylococcus aureus (MRSA) is a major hospital- and community-acquired pathogen, but the mechanisms underlying host-defense to MRSA remain poorly understood. Here, we investigated the role of IL-21 in this process. When administered intra-tracheally into wild-type mice, IL-21 induced granzymes and augmented clearance of pulmonary MRSA but not when neutrophils were depleted or a granzyme B inhibitor was added. Correspondingly, IL-21 induced MRSA killing by human peripheral blood neutrophils. Unexpectedly, however, basal MRSA clearance was also enhanced when IL-21 signaling was blocked, both in Il21r KO mice and in wild-type mice injected with IL-21R-Fc fusion-protein. This correlated with increased type I interferon and an IFN-related gene signature, and indeed anti-IFNAR1 treatment diminished MRSA clearance in these animals. Moreover, we found that IFNβ induced granzyme B and promoted MRSA clearance in a granzyme B-dependent fashion. These results reveal an interplay between IL-21 and type I IFN in the innate immune response to MRSA.

Comparison of Effectiveness of Germania Honey Compared to Manuka Honey in Methicillin-Resistant Staphylococcus aureus (MRSA) Killing

Purpose: Manuka honey is currently used in medical-grade sterile wound treatment products and has been shown to be effective in methicillin-resistant Staphylococcus aureus (MRSA) killing in vitro and in wound healing in a number of case studies and series. Locally produced honey in Pakistan and Chile have been proposed to be as effective as Manuka honey in bacterial killing in vitro, presenting potentially more accessible and affordable alternatives. In this study, we compared the effectiveness of a local Germania honey from Saudi Arabia to Manuka honey MGO 550 for in vitro killing of MRSA. Methodology: Overnight Muller Hinton broth cultures of 50 wound culture isolates of MRSA from 50 patients were incubated with a series of dilutions of Manuka honey MGO 550 and corresponding Germania honey dilutions for 24 h. Turbidity was assessed to determine whether bacterial growth had occurred, and no growth was confirmed by a further 24 h sub-culture on blood agar. Results/Key findings: Manuka honey MGO 550 was significantly more effective than Germania honey at MRSA killing at 100% v/v, 50% v/v and 25% v/v (p=0.025, 0.000265, and 0.000112 respectively) Conclusion: Manuka honey MGO 550 is significantly more effective in killing MRSA in vitro than Germania honey. Germania honey does not appear to be a promising locally produced alternative to Manuka honey for the development of honey-based wound dressings. Further experiments could determine if Germania honey is effective against other bacterial species.

Photo-inactivation and efflux pump inhibition of methicillin resistant Staphylococcus aureus using thiolated cobalt doped ZnO nanoparticles

Multidrug resistance (MDR) in bacteria is a major concern these days. One of the reasons is the mutation in efflux pump that prevents the retention of antibiotics and drugs in the bacterial cell. The current work is a step to overcome MDR in bacteria via inhibition of efflux pump and further photoinhibition by thiolated chitosan coated cobalt doped zinc oxide nanoparticles (Co-ZnO) in visible light. Co-ZnO were synthesized in a size range of 40–60 nm. Antibacterial activity of the Co-ZnO against methicillin resistant Staphylococcus aureus (MRSA) was found 100% at a concentration of 10 μg/ml upon activation in sunlight for 15 min. Interestingly, it was found that cobalt as a dopant was able to increase the photodynamic and photothermal activity of Co-ZnO, as in dark conditions, there was only 3–5% of inhibition at 10 μg/ml of nanoparticle concentration. Upon excitation in light, these nanoparticles were able to generate reactive oxygen species (ROS) with a quantum yield of 0.23 ± 0.034. The nanoparticles were also generating heat, Because of the magnetic nature, thus helping in more killing. Thiolated chitosan further helped in blocking the efflux pump of MRSA. The current nanoparticles were also found biocompatible on human red blood cells (LD50 = 214 μg/ml). These data suggest that the MRSA killing ability was facilitated through efflux inhibition and oxidative stress upon excitation in visible light hence, were in accordance with previous findings.

Effect of far ultraviolet light emitted from an optical diffuser on methicillin-resistant Staphylococcus aureus in vitro.

Drug-resistant bacteria such as methicillin-resistant Staphylococcus aureus (MRSA) are a target for new antimicrobial technologies. Far-UVC technology is an emerging disinfection method that directly kills microorganisms using light. In contrast with conventional UV sterilization, far-UVC light has antimicrobial capabilities without apparent harm to mammalian cells. This study examines the application of 224 nm far-UVC light delivered from a laser using an optical diffuser towards the goal of protecting against bacterial invasion around skin penetrating devices. Delivery of far-UVC using a laser and optical fibers enables exposure to unique geometries that would otherwise be shielded when using a lamp. Testing of the bactericidal potential of diffusing the far-UVC laser output over a large area was tested and yielded qualitative area killing results. The killing of MRSA using this method was also examined using an in vitro survival assay. Results followed a classic log-linear disinfection model with a rate constant of k = 0.51 cm2/mJ, which corresponds to an inactivation cross section of D90 = 4.5 mJ/cm2. This study establishes far-UVC delivered from a laser through an optical diffuser as a viable solution for disinfection of susceptible regions such as around catheters, drivelines, or other skin penetrating medical devices.

Investigating the antibacterial effect of Sca1+ lung mesenchymal stem cells on multidrug resistant bacterial species

Abstract Rationale Although bone marrow-derived mesenchymal stem cells have been extensively studied in host protection against bacterial pneumonia, the role of lung stem/mesenchymal stem cells in pneumonia caused by the multidrug-resistant bacteria remains elusive. Methods We have compared the extracellular (using 10 MOI) and intracellular (using 1 MOI) killing effects of stem cell antigen 1+ lung mesenchymal stem cells (Sca1+ LMSC), bone marrow-derived neutrophils (BMDN), and bone marrow-derived macrophages (BMDM) by infecting them with Gram positive methicillin-resistant Staphylococcus aureus (MRSA) and Gram negative carbapenem-resistant Klebsiella pneumoniae (CRKP) at different time points. We evaluated the extracellular and intracellular killing at each time point by plating them on Tryptic soy agar and MacConkey agar plates, respectively. We also have examined expression of inflammasomes or inflammasome-associated proteins in MRSA (20 MOI for 6 h) infected LMSC using Western blotting. Results Sca1+ LMSC is potent in killing (extracellular) both MRSA and CRKP at 3 h post-infection. However, BMDN did not show significant extracellular killing on MRSA and CRKP at all time points examined. Interestingly, Sca1+ LMSC showed a remarkably increased intracellular killing of MRSA than BMDM. Western blotting revealed the increased level of inflammasome and inflammasome-associated proteins (NLRP3, NLRP6 and ASC) in MRSA infected LMSC. Conclusions These findings have solidified the potential of LMSC-based therapies for pneumonia caused by MRSA and CRKP. However, in vivo (animal) studies are required to determine the effectiveness of LMSC in the clearance of these bacteria and modulation of innate immune responses in the host.

Target-oriented photofunctional nanoparticles (TOPFNs) for selective photodynamic inactivation of Methicillin-resistant Staphylococcus aureus (MRSA)

To inactivate methicillin-resistant Staphylococcus aureus (MRSA) with minimum damage to host cells and tissue, target-oriented photofunctional nanoparticles (TOPFNs) were fabricated and characterized. MRSA is a predominant infective pathogen even in hospital and non-hospital environments due to its ability to develop high levels of resistance to several classes of antibiotics through various pathways. To solve this major problem, photodynamic inactivation (PDI) method applies to treat antibiotic-resistant bacteria. PDI involves the photosensitizer (PS) and light with a specific wavelength to be able to apply for a non-invasive therapeutic procedure to treat pathogenic bacteria by inducing apoptosis or necrosis of microorganisms. However, most current PDI researches have suffered from the instability of PDI agents in the biological environment due to the lack of selectivity and low solubility of PDI agents, which leads to the low PDI efficiency. In this study, the TOPFNs were fabricated by an esterification reaction to introduce hematoporphyrin (HP) and MRSA antibody to the surface of Fe3O4 nanoparticles. The TOPFNs were designed as dispersible PDI agent in biological condition, which was effectively used for selectively capturing and killing of MRSA. The capture efficiency TOPFNs was compared with PFNs as a negative control. The results showed that the capture efficiency of TOPFNs and PFNs was 95.55% and 6.43% in MRSA and L-929 cell mixed condition, respectively. And TOPFNs have a selective killing ability for MRSA with minimum damage to L-929 cells. Furthermore, PDI effect of TOPFNs was evaluated on the mice in vivo condition in order to check the possibility of practical medical application.

Micellar chemotherapeutic platform based on a bifunctional salicaldehyde amphiphile delivers a "combo-effect" for heightened killing of MRSA.

The devastating infections caused by methicillin-resistant Staphylococcus aureus (MRSA) coupled with its high resistance towards antibiotics underscores the need for an effective anti-MRSA therapeutic. The present study illustrates the use of a salicylaldehyde based bactericidal amphiphile (C1) in generating a micellar carrier that renders delivery of therapeutic antibiotics. The inherent membrane-targeting activity of C1 present in the micelle could be leveraged to counter the resistance of MRSA and enhance cellular uptake of the released antibiotics, resulting in effective elimination of the pathogen. The inherent bactericidal and antibiofilm activity of C1 was captured in FESEM analysis, solution-based assays and fluorescence microscopy. ANS-based fluorescence spectroscopy indicated that the critical micelle concentration (CMC) for C1 was 18.5 μM in water. DLS studies and FESEM analysis indicated that the average particle size for micelles based on C1 (C1M) and rifampicin-loaded C1M (C1M- R) was smaller than vancomycin-loaded C1M (C1M- V). C1M- R and C1M- V rendered sustained release of the antibiotics in physiologically relevant fluids. Notably, following interaction with MRSA for 3 h, the relative anti-MRSA activity of C1M- R and C1M- V was nearly 12-fold and 8-fold higher, respectively, as compared to the free antibiotics at equivalent concentration, highlighting the merit of leveraging the activity of C1 and the antibiotic concurrently in the micellar system. The relative cell-free antibiotic was also manifold lower in the case of C1M- R and C1M- V treated MRSA as against treatment with free antibiotics, suggesting that the amphiphilic warhead breached the membrane barrier and enhanced cellular uptake of the released antibiotics. Interestingly, C1M- R and C1M- V exhibited a high therapeutic index, being non-toxic to HEK 293 cells at concentrations higher than their minimum inhibitory concentration (MIC) against MRSA and they could be employed as an antibacterial coating to prevent MRSA biofilm formation on surgical silk sutures. The antibiotic-replete biocompatible micelles based on a self-assembling membrane-targeting amphiphile described herein represent a promising framework to integrate multiple warheads and generate a potent anti-MRSA therapeutic material.

Multifunctional Biochar for Highly Efficient Capture, Identification, and Removal of Toxic Metals and Superbugs from Water Samples.

According to the World Health Organization, more than two billion people in our world use drinking water sources which are not free from pathogens and heavy metal contamination. Unsafe drinking water is responsible for the death of several millions in the 21st century. To find facile and cost-effective routes for developing multifunctional materials, which has the capability to resolve many of the challenges associated with drinking water problem, here, we report the novel design of multifunctional fluorescence-magnetic biochar with the capability for highly efficient separation, identification, and removal of pathogenic superbugs and toxic metals from environmental water samples. Details of synthesis and characterization of multifunctional biochar that exhibits very good magnetic properties and emits bright blue light owing to the quantum confinement effect are reported. In our design, biochar, a carbon-rich low-cost byproduct of naturally abundant biomass, which exhibits heterogeneous surface chemistry and strong binding affinity via oxygen-containing group on the surface, has been used to capture pathogens and toxic metals. Biochar dots (BCDs) of an average of 4 nm size with very bright photoluminescence have been developed for the identification of pathogens and toxic metals. In the current design, magnetic nanoparticles have been incorporated with BCDs which allow pathogens and toxic metals to be completely removed from water after separation by an external magnetic field. Reported results show that owing to the formation of strong complex between multifunctional biochar and cobalt(II), multifunctional biochar can be used for the selective capture and removal of Co(II) from environmental samples. Experimental data demonstrate that multifunctional biochar can be used for the highly efficient removal of methicillin-resistant Staphylococcus aureus (MRSA) from environmental samples. Reported results also show that melittin, an antimicrobial peptide-attached multifunctional biochar, has the capability to completely disinfect MRSA superbugs after magnetic separation. A possible mechanism for the selective separation of Co(II), as well as separation and killing of MRSA, has been discussed.