Optimizing the Bacteriostatic and Cytocompatibility Properties of Poly(hexamethylene guanidine) Hydrochloride (PHMG) via the Guanidine/Alkane Ratio.

The emergence of "superbugs" is not only problematic and potentially lethal for infected subjects but also poses serious challenges for the healthcare system. Although existing antibacterial agents have been effective in some cases, the side effects and biocompatibility generally present difficulties. The development of new antibacterial agents is therefore urgently required. In this work, we have adapted a strategy for the improvement of poly(hexamethylene guanidine) hydrochloride (PHMG), a common antibacterial agent. This involves copolymerization of separate monomer units in varying ratios to find the optimum ratio of the hydrocarbon to guanidine units for antibacterial activity. A series of these copolymers, designated as PGB, was synthesized. By varying the guanidine/hydrophobic ratio and the copolymer molecular weight, a structure-optimized PGB was identified that showed broad-spectrum antibacterial activity and excellent biocompatibility in solution. In an antibacterial assay, the copolymer with the optimum composition (hydrophobic unit content 25%) inhibited >99% Staphylococcus aureus and was compatible with mammalian cells. A polyurethane emulsion containing this PGB component formed transparent, flexible films (PGB-PU films) on a wide range of substrate surfaces, including soft polymers and metals. The PGB-PU films showed excellent bacteriostatic efficiency against nosocomial drug-resistant bacteria, such as Pseudomonas aeruginosa and methicillin-resistant S. aureus (MRSA). It is concluded that our PGB polymers can be used as bacteriostatic agents generally and in particular for the design of antibacterial surfaces in medical devices.

"Hearing Loss" in QCM Measurement of Protein Adsorption to Protein Resistant Polymer Brush Layers.

Accurate quantification of nonspecific protein adsorption on biomaterial surfaces is essential for evaluation of their antifouling properties. The quartz crystal microbalance (QCM) is an acoustic sensor widely used for the measurement of protein adsorption. However, although the QCM is highly sensitive, it does have performance limitations when working with surfaces modified with thick viscous layers. In the case of polymer brush surfaces, factors such as the thickness and viscosity of the brush may bring such limitations. In the present work, three types of antifouling molecules were used to explore the applicability of QCM for the evaluation of the protein resistance of hydrophilic polymer brush surfaces. Adsorption was also measured by surface plasmon resonance (SPR) as a reference. It was shown that the detection of adsorbed protein requires that protein be located within a critical distance from the QCM chip surface, determined by the viscosity of polymer brush. For larger proteins like fibrinogen, adsorption is expected to occur mainly "on top" of the polymer brush, and brush thickness determines whether protein is located in the "detectable zone". For smaller proteins like lysozyme, adsorption is expected to occur mainly at the chip surface and within the polymer brush layer and to be detectable by QCM. However, the quantity of adsorbed lysozyme may be underestimated when secondary adsorption also occurred. It is concluded that QCM data suggesting very low protein adsorption on polymer brush surfaces should take account of these considerations and should be treated generally with caution.

125I-radiolabeling, surface plasmon resonance, and quartz crystal microbalance with dissipation: three tools to compare protein adsorption on surfaces of different wettability.

The extent of protein adsorption is an important consideration in the biocompatibility of biomaterials. Various experimental methods can be used to determine the quantity of protein adsorbed, but the results usually differ. In the present work, self-assembled monolayers (SAMs) were used to prepare a series of model gold surfaces varying systematically in water wettability, from hydrophilic to hydrophobic. Three commonly used methods, namely, surface plasmon resonance (SPR), quartz crystal microbalance with dissipation (QCM-D), and (125)I-radiolabeling, were employed to quantify fibrinogen (Fg) adsorption on these surfaces. This approach allows a direct comparison of the mass of Fg adsorbed using these three techniques. The results from all three methods showed that protein adsorption increases with increasing surface hydrophobicity. The increase in the mass of Fg adsorbed with increasing surface hydrophobicity in the SPR data was parallel to that from (125)I-radiolabeling, but the absolute values were different and there does not seem to be a "universally congruent" relationship between the two methods for surfaces with varying wettability. For QCM-D, the variation in protein adsorption with wettability was different from that for SPR and radiolabeling. On the more hydrophobic surfaces, QCM-D gave an adsorbed mass much higher than from the two other methods, possibly because QCM-D measures both the adsorbed Fg and its associated water. However, on the more hydrophilic surfaces, the adsorbed mass from QCM-D was slightly greater than that from SPR, and both were smaller than from (125)I-radiolabeling; this was true no matter whether the Sauerbrey equation or the Voigt model was used to convert QCM-D data to adsorbed mass.

An integrated array of microfluidic oxygenators as a neonatal lung assist device: in vitro characterization and in vivo demonstration.

A miniaturized oxygenator device that is perfused like an artificial placenta via the umbilical vessels may have significant potential to save the lives of newborns with respiratory insufficiency. Recently we presented the concept of an integrated modular lung assist device (LAD) that consists of stacked microfluidic single oxygenator units (SOUs) and demonstrated the technical details and operation of SOU prototypes. In this article, we present a LAD prototype that is designed to accommodate the different needs of term and preterm infants by permitting changing of the number of parallel-stacked microfluidic SOUs according to the actual body weight. The SOUs are made of polydimethylsiloxane, arranged in parallel, and connected though 3D-printed polymeric interconnects to form the LAD. The flow characteristics and the gas exchange properties were tested in vitro using human blood. We found that the pressure drop of the LAD increased linearly with flow rate. Gas exchange rates of 2.4-3.8 µL/min/cm(2) (0.3-0.5 mL/kg/min) and 6.4-10.1 µL/min/cm(2) (0.8-1.3 mL/kg/min) for O2 and CO2 , respectively, were achieved. We also investigated protein adsorption to provide preliminary information on the need for application of anticoagulant coating of LAD materials. Albumin adsorption, as measured by gold staining, showed that surface uptake was evenly distributed and occurred at the monolayer level (>0.2 µg/cm(2) ). Finally, we also tested the LAD under in vivo conditions using a newborn piglet model (body weight 1.65-2.0 kg). First, the effect of an arteriovenous bypass via a carotid artery-to-jugular vein shortcut on heart rate and blood pressure was investigated. Heart rate and mean arterial blood pressure remained stable for extracorporeal flow rates of up to 61 mL/kg/min (101 mL/min). Next, the LAD was connected to umbilical vessels (maximum flow rate of 24 mL/min [10.4 mL/kg/min]), and O2 gas exchange was measured under hypoxic conditions (Fi O2 = 0.15) and was found to be 3.0 µL/min/cm(2) . These results are encouraging and support the feasibility of an artificial placental design for an LAD.

Modification of polyurethane surface with an antithrombin-heparin complex for blood contact: influence of molecular weight of polyethylene oxide used as a linker/spacer.

Polyurethane (PU) was modified using isocyanate chemistry to graft polyethylene oxide (PEO) of various molecular weights (range 300-4600). An antithrombin-heparin (ATH) covalent complex was subsequently attached to the free PEO chain ends, which had been functionalized with N-hydroxysuccinimide (NHS) groups. Surfaces were characterized by water contact angle and X-ray photoelectron spectroscopy (XPS) to confirm the modifications. Adsorption of fibrinogen from buffer was found to decrease by ~80% for the PEO-modified surfaces compared to the unmodified PU. The surfaces with ATH attached to the distal chain end of the grafted PEO were equally protein resistant, and when the data were normalized to the ATH surface density, PEO in the lower MW range showed greater protein resistance. Western blots of proteins eluted from the surfaces after plasma contact confirmed these trends. The uptake of ATH on the PEO-modified surfaces was greatest for the PEO of lower MW (300 and 600), and antithrombin binding from plasma (an indicator of heparin anticoagulant activity) was highest for these same surfaces. The PEO-ATH- and PEO-modified surfaces also showed low platelet adhesion from flowing whole blood. It is concluded that for the PEO-ATH surfaces, PEO in the low MW range, specifically MW 600, may be optimal for achieving an appropriate balance between resistance to nonspecific protein adsorption and the ability to take up ATH and bind antithrombin in subsequent blood contact.

Cell adhesion on a POEGMA-modified topographical surface.

It is well known that adsorbed proteins play a major role in cell adhesion. However, it has also been reported that cells can adhere to a protein-resistant surface. In this work, the behavior of L02 and BEL-7402 cells on a protein-resistant, 3D topographical surface was investigated. The topographical gold nanoparticle layer (GNPL) surfaces were prepared by chemical gold plating, and the topography was described by roughness parameters acquired from a multiscale analysis. Both smooth Au and GNPL surfaces were modified with POEGMA polymer brushes using surface-initiated ATRP. The dry and hydrated thicknesses of POEGMA brushes on both smooth and rough surfaces were measured by AFM using a nanoindentation method. Protein adsorption experiments using (125)I radiolabeling revealed similarly low levels of protein adsorption on smooth and GNPL surfaces modified with POEGMA, thus allowing an investigation of the effects of topography on cell behavior under conditions of minimal protein adsorption. The roles of VN and FN adsorption in both L02 cells and BEL-7402 cells adhesion were investigated using cell culturing with and without a serum supplement. It was found that initial cell adhesion occurred via proteins adsorbed from the cell culture medium, whereas subsequent durable cell adhesion could be attributed to the topographical structure of the surface. Although cell spreading on protein-resistant surfaces was constrained because of the lack of adsorbed proteins, we found that cells adherent to topographical surfaces were more firmly attached and thus were more durable compared to those on smooth surfaces. In general, however, we conclude that topography is more important for cell adhesion on a protein-resistant surface.

Inhibitory effect of silicon nanowires on the polymerase chain reaction.

The effect of nanomaterials on biological reactions has received much attention. We report herein that silicon nanowires (SiNWs) inhibit the polymerase chain reaction (PCR). The inhibitory effect was found to be concentration-dependent, with a minimum inhibitory concentration of about 0.4 mg ml(-1). DNA polymerase, restriction endonucleases, lysozyme and horseradish peroxidase maintained their bioactivities after exposure to SiNWs. Also the interaction of SiNWs with primers and dNTP did not lead to decreased PCR yield. Compared to primers and dNTP, template DNA showed 4.7-10.5-fold greater adsorption on SiNWs. Template bound to SiNWs was ineffective in the PCR, whereas addition of free template to the PCR system increased the yield. The results of this work suggest that the inhibitory effect of SiNWs on the PCR was due to the selective adsorption of double-stranded DNA on SiNWs, thereby decreasing the availability of template for the reaction.

One-step surface modification strategy with composition-tunable microgels: From bactericidal surface to cell-friendly surface.

As modifiers for biomaterial surfaces, soft colloidal particles not only have good film-forming properties, but can also contribute to the function of the biomaterial via their chemical and biological properties. This general approach has proven effective for surface modification, but little is known about methods to control the properties of the colloidal particles to regulate film formation and biological function. In this work, we prepared poly (N-isopropylacrylamide) microgels (ZQP) containing both a zwitterionic component (Z) to provide anti-fouling functionality, and a quaternary ammonium salt (Q) to give bactericidal functionality. Fine-tuning of the Z and Q contents allowed the preparation of microgels over a range of particle size, size distribution, charge, and film-forming capability. The films showed anti-adhesion and contact-killing properties versus Escherichia coli (E. Coli), depending on the chemical composition. They also showed excellent cytocompatibility relative to L929 cells. A variety of microgel-coated substrates (silicon wafer, PDMS, PU, PVC) showed long-term anti-bacterial activity and resistance to chemical and mechanical treatments. It is concluded that this approach allows the preparation of effective bactericidal, cytocompatible surfaces. The properties can be fine-tuned by regulation of the microgel composition, and the method is applicable universally, i.e., independent of substrate.

Robust, anti-biofouling 2D nanogel films from poly(N-vinyl caprolactam-co-vinylimidazole) polymers.

In analogy with adsorbed protein films, we have fabricated a family of 2D nanofilms composed of poly(N-vinyl caprolactam-co-vinylimidazole) (PNVCL) nanogels. NVCL was copolymerized with 1-vinylimidazole (VIM), and then cross-linked with α,ω-dibromoalkanes with 2 to 8 carbons via quaternization to form the nanogels. The swelling ratio of the gels was precisely controlled by regulating the inter-chain spacing of the polymers at the level of the carbon atom chain length of the cross-linker. The short-chain alkanes used are relatively rigid and their dimensions provide an accurate estimate of the chain spacing in the nanogels. It was shown that small differences in the carbon atom number of the cross-linking agent led to significant differences in the mechanical properties of the nanogels, in particular in the softness, deformability, and contact area (in film form), all of which increased with increasing carbon number. Films of the softer gels not only showed good adhesion to a number of substrates, but were also mechanically robust. In addition, the films showed excellent light transmission and nontoxicity to L929 cells. Nanogels of intermediate softness were shown to inhibit the adhesion of bacteria and human umbilical vein smooth muscle cells (HUVSMCs), and to be resistant to the adsorption of the plasma protein fibrinogen, indicating strong anti-biofouling properties. Gels that were either too stiff or too soft showed somewhat weaker anti-fouling activity in terms both of HUVSMCs adhesion and protein adsorption.

Ultrahigh Efficiency and Minimalist Intracellular Delivery of Macromolecules Mediated by Latent-Photothermal Surfaces.

Intracellular delivery of exogenous macromolecules by photothermal methods is still not widely employed despite its universal and clear effect on cell membrane rupture. The main causes are the unsatisfactory delivery efficiency, poor cell activity, poor cell harvest, and sophisticated operation; these challenges stem from the difficulty of simply controlling laser hotspots. Here, we constructed latent-photothermal surfaces based on multiwall carbon nanotube-doped poly(dimethyl siloxane), which can deliver cargoes with high delivery efficiency and cell viability. Also, cell release and harvest efficiencies were not affected by coordinating the hotspot content and surface structure. This system is suitable for use with a wide range of cell lines, including hard-to-transfect types. The delivery efficiency and cell viability were shown to be greater than 85 and 80%, respectively, and the cell release and harvest efficiency were greater than 95 and 80%, respectively. Moreover, this system has potential application prospects in the field of cell therapy, including stem cell neural differentiation and dendritic cell vaccines.

Dual-Functional Surfaces Based on an Antifouling Polymer and a Natural Antibiofilm Molecule: Prevention of Biofilm Formation without Using Biocides.

Pathogenic biofilms formed on the surfaces of implantable medical devices and materials pose an urgent global healthcare problem. Although conventional antibacterial surfaces based on bacteria-repelling or bacteria-killing strategies can delay biofilm formation to some extent, they usually fail in long-term applications, and it remains challenging to eradicate recalcitrant biofilms once they are established and mature. From the viewpoint of microbiology, a promising strategy may be to target the middle stage of biofilm formation including the main biological processes involved in biofilm development. In this work, a dual-functional antibiofilm surface is developed based on copolymer brushes of 2-hydroxyethyl methacrylate (HEMA) and 3-(acrylamido)phenylboronic acid (APBA), with quercetin (Qe, a natural antibiofilm molecule) incorporated via acid-responsive boronate ester bonds. Due to the antifouling properties of the hydrophilic poly(HEMA) component, the resulting surface is able to suppress bacterial adhesion and aggregation in the early stages of contact. A few bacteria are eventually able to break through the protection of the anti-adhesion layer leading to bacterial colonization. In response to the resulting decrease in the pH of the microenvironment, the surface could then release Qe to interfere with the microbiological processes related to biofilm formation. Compared to bactericidal and anti-adhesive surfaces, this dual-functional surface showed significantly improved antibiofilm performance to prevent biofilm formation involving both Gram-negative Pseudomonas aeruginosa and Gram-positive Staphylococcus aureus for up to 3 days. In addition, both the copolymer and Qe are negligibly cytotoxic, thereby avoiding possible harmful effects on adjacent normal cells and the risk of bacterial resistance. This dual-functional design approach addresses the different stages of biofilm formation, and (in accordance with the growth process of the biofilm) allows sequential activation of the functions without compromising the viability of adjacent normal cells. A simple and reliable solution may thus be provided to the problems associated with biofilms on surfaces in various biomedical applications.

An X-ray spectromicroscopy study of protein adsorption to polystyrene-poly(ethylene oxide) blends.

Synchrotron-based X-ray photoemission electron microscopy (X-PEEM) and atomic force microscopy (AFM) were used to characterize the composition and surface morphology of thin films of a polystyrene-poly(ethylene oxide) blend (PS-PEO), spun cast from dichloromethane at various mass ratios and polymer concentrations. X-PEEM reveals incomplete segregation with ∼30% of PS in the PEO region and vice versa. Protein (human serum albumin) adsorption studies show that this partial phase separation leads to greater protein repellency in the PS region, whereas more protein is detected in the PEO region compared to control samples.

Protein adsorption and cell adhesion/detachment behavior on dual-responsive silicon surfaces modified with poly(N-isopropylacrylamide)-block-polystyrene copolymer.

Diblock copolymer grafts covalently attached to surfaces have attracted considerable attention because of their special structure and novel properties. In this work, poly(N-isopropylacrylamide)-block-polystyrene (PNIPAAm-b-PS) brushes were prepared via surface-initiated consecutive atom-transfer radical polymerization on initiator-immobilized silicon. Because of the inherent thermosensitivity of PNIPAAm and the hydrophobicity difference between the two blocks, the modified surfaces were responsive to both temperature and solvent. Moreover, the diblock copolymer brushes exhibited both resistance to nonspecific protein adsorption and unique cell interaction properties. They showed strong protein resistance in both phosphate-buffered saline and blood plasma. In particular, fibrinogen adsorption from plasma at either room temperature or body temperature was less than 8 ng/cm(2), suggesting that the surfaces might possess good blood compatibility. In addition, the adhesion and detachment of L929 cells could be "tuned", and the ability to control the detachment of cells thermally was restored by block polymerization of hydrophobic, cell-adhesive PS onto a thicker PNIPAAm layer. In addition to providing a simple and effective design for advanced cell-culture surfaces, these results suggest new biomedical applications for PNIPAAm.

Precise regulation of particle size of poly(N-isopropylacrylamide) microgels: Measuring chain dimensions with a "molecular ruler".

HYPOTHESIS: Poly(N-isopropylacrylamide) microgels are used extensively in the design of drug carriers, surfaces for control of cell adhesion, and optical devices. Particle size is a key factor and has a significant influence in many areas. EXPERIMENTS: In this work, precise control of the particle size of poly(N-isopropylacrylamide) microgels was achieved by controlling the separation distance of the poly(N-isopropylacrylamide) chains. Dibromoalkanes of different size were used as an adjustable "molecular ruler" to measure molecular dimensions in poly(N-isopropylacrylamide) nanoaggregates at the critical crosslinking temperature. FINDINGS: We find that the chain separation distance decreases as the temperature increases with a sharp decrease over the 55-to-65 °C interval. Based on the observed relationships between chain separation and crosslinker, the particle size of poly(N-isopropylacrylamide) microgels can be regulated by changing the length of the "molecular ruler" (crosslinker) at the same temperature. Furthermore, for partly crosslinked poly(N-isopropylacrylamide) microgels that contain free crosslinkable sites, the particle size can be reduced still more by further crosslinking ("re-crosslinking") with crosslinkers of different size. It is shown that the particle size can be regulated by adjusting the length of "molecular ruler" and the degree of crosslinking. This work provides a "molecular level" method for precise control of poly(N-isopropylacrylamide) microgel particle size.

Dual Pathway for Promotion of Stem Cell Neural Differentiation Mediated by Gold Nanocomposites.

The neural differentiation of embryonic stem cells (ESCs) is of great value in the treatment of neurodegenerative diseases. On the basis of the two related signaling pathways that direct the neural differentiation of ESCs, we used gold nanoparticles (GNP) as a means of combining chemical and physical cues to trigger the neurogenic differentiation of stem cells. Neural differentiation-related functional units (glyco and sulfonate units on glycosaminoglycans, GAG) were anchored on the GNP surface and were then transferred to the cell membrane surface via GNP-membrane interactions. The functional units were able to activate the GAG-related signaling pathway, in turn promoting differentiation and maturation of stem cells into neuronal lineages. In addition, using the photothermal effect of GNP, the differentiation-inducing factor retinoic acid (RA), could be actively delivered into cells via laser irradiation. The RA-related intracellular signaling pathway was thereby further triggered, resulting in strong promotion of neurogenesis with a 300-fold increase in mature neural marker expression. The gold nanocomposites developed in this work provide the basis for a new strategy directing ESCs differentiation into nerve cells with high efficiency and high purity by acting on two related signaling pathways.

A Pumpless Microfluidic Neonatal Lung Assist Device for Support of Preterm Neonates in Respiratory Distress.

Premature neonates suffer from respiratory morbidity as their lungs are immature, and current supportive treatment such as mechanical ventilation or extracorporeal membrane oxygenation causes iatrogenic injuries. A non-invasive and biomimetic concept known as the "artificial placenta" (AP) would be beneficial to overcome complications associated with the current respiratory support of preterm infants. Here, a pumpless oxygenator connected to the systemic circulation supports the lung function to relieve respiratory distress. In this paper, the first successful operation of a microfluidic, artificial placenta type neonatal lung assist device (LAD) on a newborn piglet model, which is the closest representation of preterm human infants, is demonstrated. This LAD has high oxygenation capability in both pure oxygen and room air as the sweep gas. The respiratory distress that the newborn piglet is put under during experimentation, repeatedly and over a significant duration of time, is able to be relieved. These findings indicate that this LAD has a potential application as a biomimetic artificial placenta to support the respiratory needs of preterm neonates.

Imaging hydrated albumin on a polystyrene-poly(methyl methacrylate) blend surface with X-ray spectromicroscopy.

Human serum albumin (HSA) adsorbed to thin films of phase-segregated polystyrene (PS)-poly(methyl methacrylate) (PMMA) was examined under hydrated and dry environments with scanning transmission X-ray microscopy (STXM). Quantitative mapping of the protein and polymer components at 30 nm spatial resolution was achieved using near-edge X-ray absorption fine structure (NEXAFS) spectral contrast at the C 1s edge. Under fully hydrated conditions (0.005 mg/mL HSA), adsorbed HSA thicknesses in excess of its crystallographic dimensions suggest bilayer adsorption to the polar PMMA regions. Upon washing, these loosely bound protein molecules adsorbed to PMMA were removed. Upon drying, the thickness of HSA on the nonpolar PS region decreased by approximately 40%, indicative of conformational changes. It is suggested that this change occurs due to the free energy gain from the ability of the protein to unfold on the less crowded PS surface.

X-ray spectromicroscopy study of protein adsorption to a polystyrene-polylactide blend.

Synchrotron-based X-ray photoemission electron microscopy (X-PEEM) was used to study the adsorption of human serum albumin (HSA) to polystyrene-polylactide (40:60 PS-PLA, 0.7 wt %) thin films, annealed under various conditions. The rugosity of the substrate varied from 35 to 90 nm, depending on the annealing conditions. However, the characteristics of the protein adsorption (amounts and phase preference) were not affected by the changes in topography. The adsorption was also not changed by the phase inversion which occurred when the PS-PLA substrate was annealed above T(g) of the PLA. The amount of protein adsorbed depended on whether adsorption took place from distilled water or phosphate buffered saline solution. These differences are interpreted as a result of ionic strength induced changes in the protein conformation in solution.

The blood compatibility challenge. Part 2: Protein adsorption phenomena governing blood reactivity.

The adsorption of proteins is the initiating event in the processes occurring when blood contacts a "foreign" surface in a medical device, leading inevitably to thrombus formation. Knowledge of protein adsorption in this context has accumulated over many years but remains fragmentary and incomplete. Moreover, the significance and relevance of the information for blood compatibility are not entirely agreed upon in the biomaterials research community. In this review, protein adsorption from blood is discussed under the headings "agreed upon" and "not agreed upon or not known" with respect to: protein layer composition, effects on coagulation and complement activation, effects on platelet adhesion and activation, protein conformational change and denaturation, prevention of nonspecific protein adsorption, and controlling/tailoring the protein layer composition. STATEMENT OF SIGNIFICANCE: This paper is part 2 of a series of 4 reviews discussing the problem of biomaterial associated thrombogenicity. The objective was to highlight features of broad agreement and provide commentary on those aspects of the problem that were subject to dispute. We hope that future investigators will update these reviews as new scholarship resolves the uncertainties of today.

Rapid antibacterial effect of sunlight-exposed silicon nanowire arrays modified with Au/Ag alloy nanoparticles.

The continuing emergence of antibiotic-resistant bacteria due to the excessive use of antibiotics has produced a strong demand for novel strategies and new materials that do not lead to bacterial resistance. In the present work silicon nanowire arrays modified with gold-silver alloy nanoparticles (SN-Au/Ag) was investigated as a photo-induced antibacterial material. It was shown that SN-Au/Ag can kill bacteria with high efficiency under sunlight in times of the order of a few minutes, and this is achieved through synergism between photothermal and photocatalytic effects. It appears that the combined effect of heat and reactive oxygen species (ROS) causes bacteria killing through damage to the cell membrane and leakage of cytoplasm contents. Both gold and silver in the alloy nanoparticles are required for the observed bactericidal action. Moreover, the SN-Au/Ag material can be "recycled" without loss of bactericidal activity. It is concluded that the silicon nanowire arrays modified with gold-silver alloy nanoparticles developed in this work has promise as an antibacterial nanomaterial for the development of novel antibiotics.