Combined release of LL37 peptide and zinc ion from a mussel-inspired coating on porous titanium for infected bone defect repairing.

Implant-associated infections impose great burden on patient health and public healthcare. Antimicrobial peptides and metal ions are generally incorporated onto implant surface to deter bacteria colonization. However, it is still challenging to efficiently prevent postoperative infections at non-cytotoxic dosages. Herein, a scaffold based on porous titanium coated with a mussel-inspired dual-diameter TiO2 nanotubes is developed for loading dual drugs of LL37 peptide and Zn2+ with different sizes and characteristics. Benefiting from in-situ formed polydopamine layer and dual-diameter nanotubular structure, the scaffold provides an efficient platform for controllable drugs elution: accelerated release under acidic condition and sustained release for up to 28 days under neutral/alkalescent circumstances. Such combination of dual drugs simultaneously enhanced antibacterial efficacy and osteogenesis. In antibacterial test, LL37 peptide serving as bacteria membrane puncture agent, and Zn2+ acting as ROS generator, cooperatively destroyed bacterial membrane integrity and subsequently damaged bacterial DNA, endowing dual-drug loaded scaffold with remarkable bactericidal efficiency of > 92 % in vitro and > 99 % in vivo. Noteworthily, dual-drug loaded scaffold promoted bone-implant osteointegration under infectious microenvironment, overmatching single-drug load ones. It provides a promising strategy on surface modification of implant for infected bone defect repairing.

Fabrication of reversible bacteria-killing and bacteria-releasing cotton fabrics with anti-bacteria adhesion capacity for potential application in reusable medical materials.

Nowadays, more and more smart antibacterial materials have been prepared to meet some specific application area, and most of these materials have complex fabrication processes or incompatible biocompatibility. In this paper, a smart monomer that can switch between the form of quaternary ammonium salt and zwitterionic betaine was prepared and grafted onto cotton fabric. This finished cotton was smart too, it had nice antibacterial performance (99.89 % for E. coli and 99.97 % for S. aureus) in the form of quaternary ammonium salt, and it could release most of the attached bacteria when transferred to the form of zwitterionic betaine in PBS, and the form of zwitterionic betaine could converse back to the state of quaternary ammonium salt in HAC. Simultaneously, it was biocompatible in the form of zwitterionic betaine form. Furthermore, this smart material had nice function reproducibility after repeated transformations. In general, the smart antibacterial cotton could switch between bacteria-killing and bacteria-releasing reversibly, and had good biocompatibility and nice reproducibility, showing a potential application in reusable medical protective materials.

Leveraging the Dielectric Barrier Discharge Plasma Process to Create Regenerative Biocidal ePTFE Membranes.

The utilization of dielectric barrier discharge (DBD) plasma treatment for modifying substrate surfaces constitutes an easy and simple approach with a potential for diverse applications. This technique was used to modify the surface of a commercial porous expanded poly(tetrafluoroethylene) (ePTFE) film with either dimethylaminoethyl methacrylate (DMAEMA) or (trimethylamino)ethyl methacrylate chloride (TMAEMA) monomers, aiming to obtain antibacterial ePTFE. Physicochemical analyses of the membranes revealed that DBD successfully enhanced the surface energy and surface charge of the membranes while maintaining high porosity (>75%) and large pore size (>1.0 µm). Evaluation of the bacteria killing-releasing (K-R) function revealed that both DMAEMA and TMAEMA endowed ePTFE with the ability to kill Escherichia coli bacteria. However, only TMAEMA-grafted ePTFE allowed for the release of dead bacteria from the surface upon washing with sodium hexametaphosphate (SHMP) saline solution, owing to its cationic charge derived from the quaternary amine. Washing with SHMP disturbed the electrostatic force between the polymer brushes and dead bacteria, which caused the release of the dead bacteria. Lastly, dead-end bacteria filtration showed that the TMAEMA-grafted ePTFE was able to kill 99.78% of the bacteria, while approximately 61.55% of bacteria were killed upon contact. The present findings support the feasibility of using DBD plasma treatment for designing surfaces that target bacteria and aid in the containment of disease-causing pathogens.

Reversible bacteria-killing and bacteria-releasing cotton fabric with anti-bacteria adhesion ability for potential sustainable protective clothing applications.

Multifunctional antibacterial surfaces are playing an essential role in various areas. Smart antibacterial materials equipped with switchable "bacteria-killing" and "bacteria-releasing" abilities have been created by scientists. However, most of them are either biologically incompatible, or complex fabricating procedures, or cannot prevent themselves from being attached by bacteria. In this work, a double-layer smart antibacterial surface was created easily by simple surface initiate atom transfer radical polymerization: the upper layer PSBMA provides anti-bacteria adhesion capacity, the NCl bond can show bacteria-killing ability and the under layer PNIPAM can exhibit bacteria-releasing property. Remarkably, the NCl bond can interconvert with the NH bond easily, which allows switching between bacteria-killing and bacteria-releasing. As a result, the functional cotton fabrics can resist about 99.66 % of bacteria attaching, kill nearly 100 % of attached bacteria after 5 min contacting and release about 99.02 % of the formerly attached bacteria. Furthermore, the functional cotton fabric kept excellent anti-bacteria adhesion ability (about 99.27 %) and bacteria-releasing capacity (about 98.30 %) after 9 cycles of re-chlorination. In general, a reversible "bacteria-killing" and "bacteria-releasing" cotton fabric was fabricated with well anti-bacteria adhesion capacity in a simple way, and this smart multifunctional cotton fabric shows a great potential application in reusable protective clothing.

pH/NIR-responsive and self-healing coatings with bacteria killing, osteogenesis, and angiogenesis performances on magnesium alloy.

Although biodegradable polymer coatings can impede corrosion of magnesium (Mg)-based orthopedic implants, they are prone to excessive degradation and accidental scratching in practice. Bone implant-related infection and limited osteointegration are other factors that adversely impact clinical application of Mg-based biomedical implants. Herein, a self-healing polymeric coating is constructed on the Mg alloy together with incorporation of a stimuli-responsive drug delivery nanoplatform by a spin-spray layer-by-layer (SSLbL) assembly technique. The nanocontainers are based on simvastatin (SIM)-encapsulated hollow mesoporous silica nanoparticles (S@HMSs) modified with polydopamine (PDA) and polycaprolactone diacrylate (PCL-DA) bilayer. Owing to the dynamic reversible reactions, the hybrid coating shows a fast, stable, and cyclical water-enabled self-healing capacity. The antibacterial assay indicates good bacteria-killing properties under near infrared (NIR) irradiation due to synergistic effects of hyperthermia, reactive oxygens species (ROS), and SIM leaching. In vitro results demonstrate that NIR laser irradiation promotes the cytocompatibility, osteogenesis, and angiogenesis. The coating facilitates alkaline phosphatase activity and expedites extracellular matrix mineralization as well as expression of osteogenesis-related genes. This study reveals a useful strategy to develop multifunctional coatings on bioabsorbable Mg alloys for orthopedic implants.

Tattooing as a phenotypic gambit.

OBJECTIVES: Tattooing is not an evolved behavior, but it may be a phenotypic gambit to highlight immunological health. Phenotypic gambits are traits or behaviors that appear costly but occur at high rates as a honing process of natural selection not constrained by genetics. Tattooing is an ancient practice that is increasing in popularity worldwide, but it involves wounding the body, which seems counterintuitive because it challenges the immune system and makes one more susceptible to infection. But tattooing may represent a costly honest signal of fitness by "upping the ante" in an era of hygiene or a means to stimulate the immune system in a way that improves and highlights underlying fitness. MATERIALS AND METHODS: We investigated this hypothesis by assessing bacteria killing activity (BKA) in saliva samples collected during two studies of tattooing (N = 40). We compared previous tattoo experience (extent of body tattooed and hours spent being tattooed) to BKA before and after getting a new tattoo. RESULTS: Tattoo experience positively predicts post-tattoo BKA (ß = 0.48, p = 0.01), suggesting that people with more tattoo experience have a relatively more immediate and active immune response than those with less tattoo experience. DISCUSSION: Tattoo experience may elevate innate immunological vigilance, which could aid in protecting against future dermal insults.

One-step synthesized antimicrobial peptide-functionalized gold nanoclusters for selective imaging and killing of pathogenic bacteria.

The development of multifunctional nanomaterials with bacterial imaging and killing activities is of great importance for the rapid diagnosis and timely treatment of bacterial infections. Herein, peptide-functionalized gold nanoclusters (CWR11-AuNCs) with high-intensity red fluorescence were successfully synthesized via a one-step method using CWR11 as a template and by optimizing the ratio of CWR11 to HAuCl4, reaction time, pH, and temperature. The CWR11-AuNCs bound to bacteria and exhibited selective fluorescence microscopy imaging properties, which is expected to provide a feasible method for locating and imaging bacteria in complex in vivo environments. In addition, CWR11-AuNCs not only retained the antibacterial and bactericidal activities of CWR11 but also exhibited certain inhibitory or killing effects on gram-negative and gram-positive bacteria and biofilms. The MICs of CWR11-AuNCs against Escherichia coli and Staphylococcus aureus were 178 and 89 µg/ml, respectively. Surprisingly, cell viability in the CWR11-AuNC-treated group was greater than that in the CWR11-treated group, and the low cytotoxicity exhibited by the CWR11-AuNCs make them more promising for clinical applications.

Antibacterial surfaces: Strategies and applications.

Antibacterial surfaces are surfaces that can resist bacteria, relying on the nature of the material itself. It is significant for safe food and water, human health, and industrial equipment. Biofilm is the main form of bacterial contamination on the material surface. Preventing the formation of biofilm is an efficient way to develop antibacterial surfaces. The strategy for constructing the antibacterial surface is divided into bacteria repelling and bacteria killing based on the formation of the biofilm. Material surface wettability, adhesion, and steric hindrance determine bacteria repelling performance. Bacteria should be killed by surface chemistry or physical structures when they are attached to a material surface irreversibly. Killing approaches are usually in the light of the cell membrane of bacteria. This review summarizes the fabrication methods and applications of antibacterial surfaces from the view of the treatment of the material surfaces. We also present several crucial points for developing long-term stability, no drug resistance, broad-spectrum, and even programable antibacterial surfaces.

Vision redemption: Self-reporting AIEgens for combined treatment of bacterial keratitis.

Bacterial keratitis (BK) is one of the most commonly leading causes of visual impairment and blindness worldwide, and suffers the risk of drug-resistant infections due to the abuse of antibiotics. Herein, we report a cationic diphenyl luminogen with aggregation-induced emission called IQ-Cm containing isoquinolinium and coumarin units for theranostic study of BK. IQ-Cm has no obvious cytotoxicity to mammalian cells below a certain concentration, and could preferentially bind to bacteria over mammalian cells. IQ-Cm can be used as a sensitive self-reporting probe to rapidly discriminate live and dead bacteria by the visual emission colors. The intrinsic dark toxicity to bacteria and generation of reactive oxygen species under light irradiation endow IQ-Cm with excellent antibacterial activity in vitro and in BK rabbit models infected with S. aureus. The present study provides a sensitive and efficient theranostic strategy for rapid discrimination of various bacterial states and the combined treatment of BK based on the intrinsic dark antibacterial activity and photodynamic therapy effect.

Universal Intraductal Surface Antifouling Coating Based on an Amphiphilic Copolymer.

Surface modification on the inner wall of medical or industrial polymeric catheters with a high length/diameter ratio is highly desired. Herein, a universal and facile method based on an amphiphilic copolymer was developed to immobilize an intraductal surface antifouling coating for a variety of polymeric catheters. A fouling-repelled thin layer was formed by swelling-driven adsorption via directly perfusing an amphiphilic copolymer [polyvinylpyrrolidone-polydimethylsiloxane-polyvinylpyrrolidone (PVP-PDMS-PVP)] solution into catheters. In this copolymer, hydrophobic PDMS was embedded into a shrinking cross-linked network of catheters; also, PVP segments migrated to the surface under driving water to form a hydrophilic antifouling coating. Moreover, because of the coordination between I2 and pyrrolidone of PVP, the copolymer-modified intraductal surface was then infused with aqueous I2 to form the PVP-I2 complex, endowing this coating with bactericidal activity. Notably, diverse catheters with arbitrary shapes (circular, rectangular, triangular, and hexagonal) and different components (silicone, polyurethane, and polyethylene) were also verified to work using this interfacial interpenetration strategy. The findings in this work provide a new avenue toward facile and universal fabrication of intraductal surface antifouling catheters, creating a superior option for decreasing the consumable costs in industrial production and alleviating the pain of replacing catheters for patients.

Membrane Interactions of Virus-like Mesoporous Silica Nanoparticles.

In the present study, we investigated lipid membrane interactions of silica nanoparticles as carriers for the antimicrobial peptide LL-37 (LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES). In doing so, smooth mesoporous nanoparticles were compared to virus-like mesoporous nanoparticles, characterized by a "spiky" external surface, as well as to nonporous silica nanoparticles. For this, we employed a combination of neutron reflectometry, ellipsometry, dynamic light scattering, and ζ-potential measurements for studies of bacteria-mimicking bilayers formed by palmitoyloleoylphosphatidylcholine/palmitoyloleoylphosphatidylglycerol. The results show that nanoparticle topography strongly influences membrane binding and destabilization. We found that virus-like particles are able to destabilize such lipid membranes, whereas the corresponding smooth silica nanoparticles are not. This effect of particle spikes becomes further accentuated after loading of such particles with LL-37. Thus, peptide-loaded virus-like nanoparticles displayed more pronounced membrane disruption than either peptide-loaded smooth nanoparticles or free LL-37. The structural basis of this was clarified by neutron reflectometry, demonstrating that the virus-like nanoparticles induce trans-membrane defects and promote incorporation of LL-37 throughout both bilayer leaflets. The relevance of such effects of particle spikes for bacterial membrane rupture was further demonstrated by confocal microscopy and live/dead assays on Escherichia coli bacteria. Taken together, these findings demonstrate that topography influences the interaction of nanoparticles with bacteria-mimicking lipid bilayers, both in the absence and presence of antimicrobial peptides, as well as with bacteria. The results also identify virus-like mesoporous nanoparticles as being of interest in the design of nanoparticles as delivery systems for antimicrobial peptides.

Maternal provisioning and fluctuating thermal regimes enhance immune response in a reptile with temperature-dependent sex determination.

The Charnov-Bull model of differential fitness is often used to explain the evolution and maintenance of temperature-dependent sex determination (TSD). Most tests of the model focus on morphological proxies of fitness, such as size traits, whereas early life physiological traits that are closely related to lifetime fitness might provide a framework for generalizing the Charnov-Bull model across taxa. One such trait is the strength of the early-life immune response, which is strongly linked to early-life survival and fitness. Here, we manipulated temperature, variance in temperature, and sex to test the Charnov-Bull model using a physiological trait, immune system strength, in the snapping turtle (Chelydra serpentina). We found no evidence of sex-specific differences in bactericidal capacity of hatchling blood, and no evidence that mean temperature influences bactericidal capacity. However, we did find that fluctuating incubation temperature (i.e. a more naturalized incubation regime) is associated with a greater bactericidal capacity compared with constant temperature incubation. We also found that egg mass, a proxy for maternal provisioning, is positively associated with bactericidal capacity. Our findings suggest that the evolution of temperature-dependent sex determination in reptiles is unrelated to our measure of early-life innate immunity. Our study also underlines how immune response is condition dependent in early life, and questions the biological relevance of constant temperature incubation in experimental studies on ectotherm development.

Effect of plasma-induced oxidative stress on the glycolysis pathway of Escherichia coli.

Antibiotic resistance is one of the world's most urgent public health problems. Due to its antibacterial properties, cold atmospheric plasma (CAP) may serve as an alternative method to antibiotics. It is claimed that oxidative stress caused by CAP is the main reason of bacteria inactivation. In this work, we computationally investigated the effect of plasma-induced oxidation on various glycolysis metabolites, by monitoring the production of the biomass. We observed that in addition to the significant reduction in biomass production, the rate of some reactions has increased. These reactions produce anti-oxidant products, showing the bacterial defense mechanism to escape the oxidative damage. Nevertheless, the simulations show that the plasma-induced oxidation effect is much stronger than the defense mechanism, causing killing of the bacteria.

The Role of Properdin in Killing of Non-Pathogenic Leptospira biflexa.

Properdin (P) is a positive regulatory protein that stabilizes the C3 convertase and C5 convertase of the complement alternative pathway (AP). Several studies have suggested that properdin can bind directly to the surface of certain pathogens regardless of the presence of C3bBb. Saprophytic Leptospira are susceptible to complement-mediated killing, but the interaction of properdin with Leptospira spp. has not been evaluated so far. In this work, we demonstrate that properdin present in normal human serum, purified properdin, as well as properdin oligomers P2, P3, and P4, interact with Leptospira. Properdin can bind directly to the bacterial surface even in the absence of C3b. In line with our previous findings, AP activation was shown to be important for killing non-pathogenic L. biflexa, and properdin plays a key role in this process since this microorganism survives in P-depleted human serum and the addition of purified properdin to P-depleted human serum decreases the number of viable leptospires. A panel of pathogenic L.interrogans recombinant proteins was used to identify putative properdin targets. Lsa30, an outer membrane protein from L. interrogans, binds to unfractionated properdin and to a lesser extent to P2-P4 properdin oligomers. In conclusion, properdin plays an important role in limiting bacterial proliferation of non-pathogenic Leptospira species. Once bound to the leptospiral surface, this positive complement regulatory protein of the AP contributes to the formation of the C3 convertase on the leptospire surface even in the absence of prior addition of C3b.

Active pharmaceutical ingredient poly(ionic liquid)-based microneedles for the treatment of skin acne infection.

As an inflammatory skin disease of pilosebaceous follicles, Propionibacterium acnes (P. acnes) can aggravate local inflammatory responses and forms acne lesions. However, due to the skin barrier, various transdermal measures other than antibiotic creams are necessary. Microneedle (MN) patches are emerging platforms for the transdermal delivery of various therapeutics since it can effectively create transport pathways in the epidermis. Herein, we develop an active pharmaceutical ingredient poly(ionic liquid) (API PIL)-based MN patches containing salicylic acid (SA). The PIL-based MNs are simply prepared through photo-crosslinking of an imidazolium-type ionic liquid (IL) monomer in MN micro-molds, and following by anion exchange with salicylic acid anions (SA-). The fabricated SA-loaded PIL-MNs exhibited therapeutic efficiency in the topical treatment of P. acnes infection in vitro and in vivo. These active pharmaceutical ingredient PIL-based MNs can improve acne treatment, demonstrating potential applications for skin diseases. STATEMENT OF SIGNIFICANCE: Microneedle (MN) patches can be used as platforms for transdermal delivery of various therapeutics to treat bacterial infection. Here, a facile strategy was developed to synthesize active pharmaceutical ingredient poly(ionic liquid)-based microneedle patches by anion-exchange with salicylic acid anion (SA-). The fabricated SA-loaded PIL-MNs are active on not only anti-bacteria but also anti-inflammation in P. acnes treated mice, and may have potential applications for skin acne infection.

Thorn-like flexible Ag2C2O4/TiO2 nanofibers as hierarchical heterojunction photocatalysts for efficient visible-light-driven bacteria-killing.

TiO2-based fibrous membranes as plasmonic heterojunction photocatalysts would hold great promise in the field of water disinfection, however, it still existed a great challenge to design and construct such materials. Here, we presented the fabrication of continuous, hierarchical, and easy-to-recycle flexible Ag2C2O4/TiO2 heterostructured nanofibrous membranes (NMs) that were composed of thorn-like nanofibers through electrospinning technique followed by successive ionic layer adsorption and reaction (SILAR) process. Ag2C2O4 nanoplates were firmly anchored on the surface of TiO2 and the obtained Ag2C2O4/TiO2 heterojunction photocatalysts underwent a silent-to-active transition of visible-light response under light irradiation due to the surface plasmon resonance (SPR) effect of Ag nanoparticles derived from Ag2C2O4, forming a new plasmonic heterojunction photocatalyst. By virtue of the hierarchical structure, enhanced visible light absorption and efficient charge carriers separation, Ag2C2O4/TiO2 NMs possessed high bactericidal efficiency of >99.999% within 30 min, strong reactive oxygen species (ROS) producing capability (1510 µg g-1 and 659 µg g-1 for superoxide radicals and hydroxyl radicals, respectively), and good reusability. This work may offer new insights into the design of antibacterial materials for pathogenic microorganism-contaminated water purification.

Egg Incubation Temperature Affects Development of Innate Immune Function in Nestling American Robins (Turdus migratorius).

The innate immune system provides important first-line defenses against invading pathogens and is considered especially important for developing organisms. However, we know little about how early-life conditions influence these defenses in wild animals. For oviparous species such as birds, embryonic development occurs in the egg, which can be subject to variation in thermal conditions. There is evidence from cavity-nesting species and species with precocial young that reduced incubation temperatures can result in reduced measures of innate immunity. Whether and how this thermal variation impacts innate immunity for open-cup-nesting species with altricial offspring has not been examined. In this study, we experimentally manipulated egg incubation temperature for American robins (Turdus migratorius) and compared the bacteria-killing ability (BKA) of the nestlings' blood plasma. We collected baseline and poststressor samples on day 7 and day 10 after hatch to gain additional insights into the ontogeny of this immune measure, as well as into whether any changes were linked to levels of the glucocorticoid hormone corticosterone (CORT). We found that nestlings incubated at the low treatment (36.1°C) had significantly reduced BKA compared with nestlings incubated at the high treatment (37.8°C) when controlling for the posthatch nest environment. We also documented a significant reduction in poststressor levels of BKA, as well as an increase in BKA from day 7 to day 10. We found a weak inverse association between CORT and BKA but no other indications that BKA was mediated via treatment-induced variation in CORT. Our results suggest that incubation temperature can affect development of innate immunity in open-cup-nesting passerines.

Bacteria killer enzyme attached magnetic nanoparticles.

The objective of the present work was to develop immobilized lysozyme systems through adsorption on magnetic nanoparticles for potential usage in bacteria killing studies. For this, magnetic poly(HEMA-GMA) nanoparticles were prepared by surfactant free emulsion polymerization technique and functionalized with dye ligand Reactive Green 5. Synthesized magnetic nanoparticles were then characterized by FTIR, SEM, EDX and ESR studies. Particle size range of the polymers was found to be as 90-120 nm. Magnetic behavior was also demonstrated by ESR with the g value of 2.48. Maximum lysozyme loading was found to be as 1045.1 mg/g nanopolymer. Repeated usability of the magnetic nanoparticles was also studied. Immobilized form of lysozyme protected 85.85% of its initial activity at the end of the immobilization process. Bacteria killing capacity of the lysozyme immobilized magnetic nanoparticles were investigated by using Micrococcus lysodeikticus bacteria and it was demonstrated that all bacteria were successfully destroyed by the lysozyme immobilized magnetic nanoparticles within 5 min.

Localization of all four ZnT zinc transporters in Dictyostelium and impact of ZntA and ZntB knockout on bacteria killing.

Professional phagocytes have developed an extensive repertoire of autonomous immunity strategies to ensure killing of bacteria. Besides phagosome acidification and the generation of reactive oxygen species, deprivation of nutrients and the lumenal accumulation of toxic metals are essential to kill ingested bacteria or inhibit the growth of intracellular pathogens. Here, we used the soil amoeba Dictyostelium discoideum, a professional phagocyte that digests bacteria for nutritional purposes, to decipher the role of zinc poisoning during phagocytosis of nonpathogenic bacteria and visualize the temporal and spatial dynamics of compartmentalized, free zinc using fluorescent probes. Immediately after particle uptake, zinc is delivered to phagosomes by fusion with 'zincosomes' of endosomal origin, and also by the action of one or more zinc transporters. We localized the four Dictyostelium ZnT transporters to endosomes, the contractile vacuole and the Golgi complex, and studied the impact of znt knockouts on zinc homeostasis. We show that zinc is delivered into the lumen of Mycobacterium smegmatis-containing vacuoles, and that Escherichia coli deficient in the zinc efflux P1B-type ATPase ZntA are killed faster than wild-type bacteria.

Measuring Neutrophil Bactericidal Activity.

The best-known role of neutrophils is control of pathogen growth. Neutrophils contain and kill pathogens through a variety of antimicrobial activities. Regardless of the mechanism, the ability to kill pathogens is a vital outcome. This chapter describes a method to measure the in vitro bactericidal activity of isolated neutrophils as the endpoint of converging innate immune functions.