Development of silver-containing austenite antibacterial stainless steels for biomedical applications Part I: microstructure characteristics,mechanical properties and antibacterial mechanisms

The as-quenched (AQ) microstructure of the Ag-containing alloys was found to be essentially a mixture of austenite (γ) and Ag phases. The Ag phase precipitates had a face-centered-cubic structure and lattice parameter a = 4.09 Å. When the alloy contained Ag ≥0.2 wt%, the mechanical properties were slightly enhanced because of the precipitate strengthening by the Ag phase precipitates. Moreover, the Ag-containing alloys exhibited ductile fracture after tensile testing. The results of an antibacterial test revealed that the Ag phase precipitates play a key role in the antibacterial mechanism of Ag-containing alloys: Ag⁺ ions released from the Ag phase precipitates can kill bacteria. It is suggested that as AISI 316L alloy has an Ag content ≥0.2 wt%, it will have excellent antibacterial properties against both Staphylococcus aureus and Escherichia coli, with an antibacterial rate of nearly 100%.     

A new concept in polymeric thin-film composite nanofiltration membranes with antibacterial properties

A new, thin film, biofouling resistant, nanofiltration (NF) membrane was fabricated with two key characteristics, viz. a low rate of silver (Ag) release and long-lasting antibacterial properties. In the new approach, nanoparticles were embedded completely in a polymeric thin-film layer. A comparison was made between the new thin-film composite (TFC), NF membrane and thin-film nanocomposite (TFN), and antibacterial NF membranes. Both types of NF membrane were fabricated by interfacial polymerization on a polysulphone sublayer using m-phenylenediamine and trimesoyl chloride as an amine monomer and an acid chloride monomer, respectively. Energy dispersive X-ray (EDX) microanalysis demonstrated the presence of Ag nanoparticles. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) were used to study the cross-sectional and surface morphological properties of the NF membranes. Permeability and salt rejection were tested using a dead-end filtration cell. Ag leaching from the membranes was measured using inductively coupled mass spectrometry (ICP–MS). Morphological studies showed that the TFC NF membranes had better thin-film formation (a more compact structure and a smoother surface) than TFN NF membranes. Performance experiments on TFC NF membranes revealed that permeability was good, without sacrificing salt rejection. The antibacterial properties of the fabricated membranes were tested using the disk diffusion method and viable plate counts. The antibiofouling properties of the membranes were examined by measuring the quantity of bacterial cells released from the biofilm formed (as a function of the amount of biofilm present). A more sensitive surface was observed compared to that of a typical antibacterial NF membrane. The Ag leaching rates were low, which will likely result in long-lasting antibacterial and biofouling resistant properties.     

Preparation of chitosan-nylon-6 blended membranes containing silver ions as antibacterial materials

Chitosan-nylon-6 blended membranes were prepared by combining solvent evaporation and a phase inversion technique, and then used to chelate silver ions. Gram-positive bacteria (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli) were used to study the antibacterial properties of the membranes. Fourier-transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC) indicated hydrogen-bond interactions between chitosan and nylon-6. From the scanning electron microscopy (SEM) pictures, it was observed that with the increase of nylon-6 content, the blended membrane gradually became a material with porous morphology. After chelating silver ions, the tensile strength of the membranes increased. The antibacterial activity with the variation of chitosan content, the pH value and the concentration of the silver nitrate solution used to prepare Ag(+)-loaded membranes were investigated systematically. The results indicated that the chitosan-nylon-6 blended membranes with Ag(+) were antibacterial to both Gram-positive bacteria and Gram-negative bacteria. The antibacterial activity improved with the increased chitosan content due to the larger amount of silver ions loaded. The antibacterial property of the chitosan-nylon-6 blended membranes could be primarily attributed to the content of chitosan and silver ions as well as the surface morphology of the membranes.     

Silver nanoparticles: partial oxidation and antibacterial activities

The physical and chemical properties of silver nanoparticles that are responsible for their antimicrobial activities have been studied with spherical silver nanoparticles (average diameter approximately 9 nm) synthesized by the borohydride reduction of Ag+ ions, in relation to their sensitivity to oxidation, activities towards silver-resistant bacteria, size-dependent activities, and dispersal in electrolytic solutions. Partially (surface) oxidized silver nanoparticles have antibacterial activities, but zero-valent nanoparticles do not. The levels of chemisorbed Ag+ that form on the particle's surface, as revealed by changes in the surface plasmon resonance absorption during oxidation and reduction, correlate well with the observed antibacterial activities. Silver nanoparticles, like Ag+ in the form of AgNO3 solution, are tolerated by the bacteria strains resistant to Ag+. The antibacterial activities of silver nanoparticles are related to their size, with the smaller particles having higher activities on the basis of equivalent silver mass content. The silver nanoparticles aggregate in media with a high electrolyte content, resulting in a loss of antibacterial activities. However, complexation with albumin can stabilize the silver nanoparticles against aggregation, leading to a retention of the antibacterial activities. Taken together, the results show that the antibacterial activities of silver nanoparticles are dependent on chemisorbed Ag+, which is readily formed owing to extreme sensitivity to oxygen. The antibacterial activities of silver nanoparticles are dependent on optimally displayed oxidized surfaces, which are present in well-dispersed suspensions.     

Factors affecting the antibacterial activity of the ruminal bacterium,Streptococcus bovis HC5

Streptococcus bovis HC5 inhibits a variety of S. bovis strains and other Gram-positive bacteria, but factors affecting this activity had not been defined. Batch culture studies indicated that S. bovis HC5 did not inhibit S. bovis JB1 (a non-bacteriocin-producing strain) until glucose was depleted and cells were entering stationary phase, but slow-dilution-rate, continuous cultures (0.2 h(-1)) had as much antibacterial activity as stationary-phase batch cultures. Because the activity of continuous cultures (0.2-1.2 h(-1)) was inversely related to the glucose consumption rate, it appeared that the antibacterial activity was being catabolite repressed by glucose. When the pH of continuous cultures (0.2 h(-1)) was decreased from 6.7 to 5.4, antibacterial activity doubled, but this activity declined at pH values less than 5.0. Continuous cultures (0.2 h(-1)) that had only ammonia as a nitrogen source had antibacterial activity, and large amounts of Trypticase (10 mg ml(-1)) caused only a 2.0-fold decline in the amount of HC5 cell-associated protein that was needed to prevent S. bovis JB1 growth. Because S. bovis HC5 was able to produce antibacterial activity over a wide range of culture conditions, there is an increased likelihood that this activity could have commercial application.     

Preparation and characterization of konjac glucomannan/poly(diallydimethylammonium chloride) antibacterial blend films

A novel antibacterial film was prepared by blending konjac glucomannan (KGM) and poly(diallydimethylammonium chloride) (PDADMAC) in an aqueous system. The antibacterial activity of the films against Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa, and Saccharomyces were measured by the halo zone test and the double plate method. The films exhibited an excellent antibacterial activity against B. subtilis and S. aureus but not against E. coli, P. aeruginosa or Saccharomyces. The miscibility, morphology, thermal stability, water vapour permeability and mechanical properties of the blend films were investigated by density determination, SEM, ATR-IR, XRD, DSC, TGA, WVA and tensile tests. The results of density determination predicted that the blends of KGM and PDADMAC were miscible when the PDADMAC content was less than 70 wt%. Moreover, SEM and XRD confirmed the result. ATR-IR showed that strong intermolecular hydrogen bonds and electrostatic interactions occurred between KGM and PDADMAC in the blends. The tensile strength and the break elongation of the blends were improved largely to 106.5 MPa and 32.04% and the water vapour permeability decreased when the PDADMAC content was 20 wt%. The thermal stability of the blends was higher than pure KGM. The blends should be good antibacterial materials.     

Synthesis of antibacterial PVA/CM-chitosan blend hydrogels with electron beam irradiation

A series of excellent hydrogels were prepared from poly(vinyl alcohol) (PVA) and carboxymethylated chitosan (CM-chitosan) with electron beam irradiation (EB) at room temperature. Electron spectroscopy analysis of the blend hydrogels revealed that good miscibility was sustained between CM-chitosan and PVA. The properties of the prepared hydrogels, such as the mechanical properties, gel fraction and swelling behavior were investigated. The mechanical properties and equilibrium degree of swelling improved obviously after adding CM-chitosan into PVA hydrogels. The gel fraction determined gravimetrically showed that a part of CM-chitosan was immobilized onto PVA hydrogel. The further analyses of FTIR and DSC spectra of the prepared gels after extracting sol manifested that there was a grafting interaction between PVA and CM-chitosan molecules under irradiation. The antibacterial activity of the hydrogels against Escherichia coli was also measured via optical density method. The blend hydrogels exhibited satisfying antibacterial activity against E. coli, even when the CM-chitosan concentration was only 3 wt%.     

Photo-catalytic preparation of silver-coated TiO2 particles for antibacterial applications

Colloidal silver has been known to have unique antimicrobial activity that may be useful in the construction of antibacterial materials (self-cleaning materials) to aid in the fight against bacteria-related infections. In this study, silver-coated TiO₂ (Ag/TiO₂) particles prepared through the photo-reduction of Ag⁺ were investigated as an antibacterial agent against Escherichia coli and Staphylococcus aureus. The deposition of Ag onto the surface was confirmed with SEM and EDS analysis of the post-reaction particles. It was also determined that the initial concentration of Ag⁺ in solution played a significant role in the effective size of the post-irradiation particles. The antibacterial effectiveness of the Ag/TiO₂ was evaluated through the determination of the minimum inhibitory concentration (MIC) of AgTiO₂ for each species of bacteria. The MIC values for the Ag/TiO₂, on both E. coli and S. aureus, were much lower than the MIC values for Ag metal, and quite comparable to the MIC values for AgNO₃. A disc diffusion/antibiotic sensitivity test was also performed using the Ag/TiO₂ particles and the results compared with the results obtained for Ag metal, AgNO₃ and common antibacterial agents; tetracycline, chloramphenicol, erythromycin, and neomycin. The zone of inhibition diameters for the Ag/TiO₂ particles were found to be comparable with those of the other antimicrobial agents.     

掺银二氧化钛空心球的制备及其抗菌性能研究

以硫酸钛、尿素为原料,碳球为模板,采用水热沉淀法制备了二氧化钛空心球,以聚乙烯吡咯烷酮(PVP)为分散剂、硼氢化钾还原硝酸银的方法得到银掺杂的二氧化钛空心球。采用X射线粉末衍射仪(XRD)、扫描电镜(SEM)、透射电镜(TEM)、能谱仪(EDS)和傅里叶变换红外光谱仪(FT-IR)等测试手段对所得的样品进行了表征。采用抑菌圈法对其抗菌性能进行了检测,实验结果表明,在自然光条件下,商品二氧化钛P25和二氧化钛空心球对大肠杆菌、金黄色葡萄球菌、白色念珠菌没有任何抗菌效果,而载银量9.4 mol%的二氧化钛在常温范围内对三个菌种都具有优良的抗菌性能,可用于抗菌塑料、抗菌食品包装和抗菌纺织制品等领域。     

Graphene oxide/silver nanohybrid: Optimization,antibacterial activity and its impregnation on bacterial cellulose as a potential wound dressing based on GO‐Ag nanocomposite‐coated BC

Recently, bacterial cellulose (BC) based wound dressing have raised significant interests in medical fields. However, to our best knowledge, it is apparent that the BC itself has no antibacterial activity. In this study, we optimized graphene oxide‐silver (GO‐Ag) nanohybrid synthesis using Response Surface Methodology and impregnate it to BC and carefully investigate their antibacterial activities against both the Gram‐negative bacteria Escherichia coli and the Gram‐positive bacteria Staphylococcus aureus. We discover that, compared to silver nanoparticles, GO‐Ag nanohybrid with an optimal GO suspension's pH and ratio is much more effective and shows synergistically enhanced, strong antibacterial activities at rather low dose. The GO‐Ag nanohybrid is more toxic to E. coli than that to S. aureus. The antibacterial and mechanical properties of BC/GO‐Ag composite are further investigated.     

Proteomic analysis of the mode of antibacterial action of silver nanoparticles

Silver nanoparticles (nano-Ag) are potent and broad-spectrum antimicrobial agents. In this study, spherical nano-Ag (average diameter = 9.3 nm) particles were synthesized using a borohydride reduction method and the mode of their antibacterial action against E. coli was investigated by proteomic approaches (2-DE and MS identification), conducted in parallel to analyses involving solutions of Ag(+) ions. The proteomic data revealed that a short exposure of E. coli cells to antibacterial concentrations of nano-Ag resulted in an accumulation of envelope protein precursors, indicative of the dissipation of proton motive force. Consistent with these proteomic findings, nano-Ag were shown to destabilize the outer membrane, collapse the plasma membrane potential and deplete the levels of intracellular ATP. The mode of action of nano-Ag was also found to be similar to that of Ag(+) ions (e.g., Dibrov, P. et al, Antimicrob. Agents Chemother. 2002, 46, 2668-2670); however, the effective concentrations of nano-Ag and Ag(+) ions were at nanomolar and micromolar levels, respectively. Nano-Ag appear to be an efficient physicochemical system conferring antimicrobial silver activities.     

Biosensitive and antibacterial coatings on metallic material for medical applications

Metallic materials are commonly used for load-bearing implants and as internal fixation devices. It is customary to use austenitic stainless steel, especially surgical grade type 316L SS as temporary and Ti alloys as permanent implants. However, long-term, poor bonding with bone, corrosion, and release of metal ions, such as chromium and nickel occur. These ions are powerful allergens and carcinogens and their uncontrolled leaching may be avoided by surface coatings. Therefore, bioactive glasses (BGs) became a vital biomedical material, which can form a biologically active phase of hydroxycarbonate apatite on their surface when in contact with physiological fluids. To reduce the high coefficient of friction and the brittle nature of BGs, polymers are normally incorporated to avoid the high-temperature sintering/densification of ceramic-only coatings. For medical application, electrophoretic deposition (EPD) is now used for polymer (organic) and ceramic (inorganic) components at room temperature due to its simplicity, control of coating thickness and uniformity, low cost of equipment, ability to coat substrates of intricate shape and to supply thick films in composite form, high purity of deposits as well as no phase transformation during coating. Although extensive research has been conducted on polymer/inorganic composite coatings, only some studies have reported multifunctional properties, such as biological antibacterial activity, enhanced cell adhesion, controlled drug release ability, and mechanical properties. This review will focus on biodegradable coatings, including zien, chitosan, gelatin, cellulose loaded with antibacterial drugs/metallic ions/natural herbs on biostable substrates (PEEK/PMMA/PCL/PLLA layers), which have the potential of multifunctional coating for metallic implants.     

Electro and Magneto-Electropolished Surface Micro-Patterning on Binary and Ternary Nitinol

In this study, an Atomic Force Microscopy (AFM) roughness analysis was performed on non-commercial Nitinol alloys with Electropolished (EP) and Magneto-Electropolished (MEP) surface treatments and commercially available stents by measuring Root-Mean-Square (RMS), Average Roughness (Ra), and Surface Area (SA) values at various dimensional areas on the alloy surfaces, ranging from (800 × 800 nm) to (115 × 115μm), and (800 × 800 nm) to (40 × 40 μm) on the commercial stents. Results showed that NiTi-Ta 10 wt% with an EP surface treatment yielded the highest overall roughness, while the NiTi-Cu 10 wt% alloy had the lowest roughness when analyzed over (115 × 115 μm). Scanning Electron Microscopy (SEM) and Energy Dispersive Spectroscopy (EDS) analysis revealed unique surface morphologies for surface treated alloys, as well as an aggregation of ternary elements Cr and Cu at grain boundaries in MEP and EP surface treated alloys, and non-surface treated alloys. Such surface micro-patterning on ternary Nitinol alloys could increase cellular adhesion and accelerate surface endothelialization of endovascular stents, thus reducing the likelihood of in-stent restenosis and provide insight into hemodynamic flow regimes and the corrosion behavior of an implantable device influenced from such surface micro-patterns.     

Nano silver entrapped in phospholipids membrane: synthesis, characteristics and antibacterial kinetics

The antimicrobial property of stabilized silver nanoparticles (AgNPs) with phospholipid membrane was investigated on both Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) bacterial strains. The influence of phospholipid concentrations on antibacterial kinetics actions of AgNPs was studied with two different methodologies in order to understand the bactericidal and bacteriostatic effects. The bacterial inactivation of synthesized AgNPs fitted well to the Chick-Watson model with a high regression coefficient, R(2) > 0.91. The antibacterial properties of AgNPs depend on the particle size, stabilizer and lecithin concentrations. Only the stabilized AgNPs that have the K(lec/Ag) values of 1 and 2 presented the inhabitation zone, while unstabilized AgNPs agglomerated quickly, settled on the wells and did not diffuse in agar. In addition, the specific coefficient of lethality depends on the lecithin concentration. An increase in lecithin concentration caused multilayer creation on the AgNPs' surface and reduced the release of AgNPs which led to low bacterial killing rate.     

纳米银胶/壳聚糖复合抗菌剂的制备及其在洗涤产品中抗菌性能

本文研究了纳米银胶/壳聚糖抗菌剂的制备及其形貌的表征分析,以大肠杆菌为代表菌株,研究了复合抗菌剂在洗涤产品中的抗菌效率及抗菌的稳定性,结果说明复合抗菌剂在洗涤产品中添加1.0%时,其抗菌效率达99%,经180 d长期分析,其抗菌活性仍保持95%左右。此外,复合抗菌剂对不同菌株的抗菌性能也均较强。     

Facts and myths of antibacterial properties of silk

Silk cocoons provide protection to silkworm from biotic and abiotic hazards during the immobile pupal phase of the lifecycle of silkworms. Protection is particularly important for the wild silk cocoons reared in an open and harsh environment. To understand whether some of the cocoon components resist growth of microorganisms, in vitro studies were performed using gram negative bacteria Escherichia coli (E. coli) to investigate antibacterial properties of silk fiber, silk gum, and calcium oxalate crystals embedded inside some cocoons. The results show that the previously reported antibacterial properties of silk cocoons are actually due to residues of chemicals used to isolate/purify cocoon elements, and properly isolated silk fiber, gum, and embedded crystals free from such residues do not have inherent resistance to E. coli. This study removes the uncertainty created by previous studies over the presence of antibacterial properties of silk cocoons, particularly the silk gum and sericin. © 2013 Wiley Periodicals, Inc. Biopolymers 101: 237–245, 2014.     

The antibacterial properties of secreted phospholipases A(2)

There is a considerable body of evidence to support the antibacterial properties of the group IIa phospholipase A(2) as an important physiological function. This enzyme is able to act as an acute phase protein and may be part of the innate defence system of the body, acting in concert with other antibacterial proteins and peptides. The enzyme is most effective against Gram-positive bacteria whereas penetration of the lipopolysaccharide coat of Gram-negative bacteria requires bactericidal/permeability-increasing protein (BPI) as an additional permeabilizing factor. The global cationic nature of this protein (pI>10.5) appears to facilitate penetration of the anionic bacterial cell wall. In addition, the considerable preference of the enzyme for anionic phospholipid interfaces provides specificity toward anionic bacterial membranes as opposed to zwitterionic eucaryotic cell membranes.     

Mechanisms of the enhanced antibacterial effect of Ag-TiO2 coatings

It has been demonstrated that Ag-TiO₂ nanocomposite coatings with excellent antimicrobial activity and biocompatibility have the potential to reduce infection problems. However, the mechanism of the synergistic effect of Ag-TiO₂ coatings on antibacterial efficiency is still not well understood. In this study, five types of Ag-TiO₂ nanocomposited coatings with different TiO₂ contents were prepared on a titanium substratum. Leaching tests indicated that the incorporation of TiO₂ nanoparticles into an Ag matrix significantly promoted Ag ion release. Surface energy measurements showed that the addition of TiO₂ nanoparticles also significantly increased the electron donor surface energy of the coatings. Bacterial adhesion assays with Escherichia coli and Staphylococcus aureus demonstrated that the number of adhered bacteria decreased with increasing electron donor surface energy. The increased Ag ion release rate and the increased electron donor surface energy contributed to an enhanced antibacterial efficiency of the coatings.     

Clotting of citrated plasma by a proteolytic enzyme associated with an antibacterial agent from serum

Two to 4 mg/ml of an antibacterial agent occurring in the serum of humans and animals caused the clotting of citrated rabbit, human, calf, ox, sheep, goat, guinea pig, rat, mouse, horse, chicken, pigeon and swine plasma. Heparinized plasmas of the same species were found resistant to the clotting action of the antibacterial agent. Citrated plasma previously heated at 56 C for 30 min, treated with 640 units/ml tyrosinase for 60 min, or absorbed with 0.2M Ca₃(PO₄)₂ and 0.2M Mg(OH)₂ was also found resistant to the clotting action of the antibacterial agent. The clotting action of the antibacterial agent was not affected by heating at 66 C for 60 min, nor by multiple passage through Seitz filters. The coagulation of citrated plasma proceeded most rapidly with 0.2M Tris(hydroxymethyl)-amino-methane buffer, pH 7 to 9, or distilled water as the antibacterial agent solvent; 0.2M phosphate buffer, pH 6 to 8, reduced the clotting action of the antibacterial agent, while 0.2M citrate-phosphate buffer, pH 4.0, or 0.2M carbonate-bicarbonate buffer, pH 10.0, inhibited entirely the clotting activity of this agent.     

A simple approach to constructing antibacterial and anti-biofouling nanofibrous membranes

In this work, antibacterial and anti-adhesive polymeric thin films were constructed on polyacrylonitrile (PAN) nanofibrous membranes in order to extend their applications. Polyhexamethylene guanidine hydrochloride (PHGH) as an antibacterial agent and heparin (HP) as an anti-adhesive agent have been successfully coated onto the membranes via a layer-by-layer (LBL) assembly technique confirmed by attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), energy-dispersive spectroscopy (EDS) and scanning electron microscopy (SEM). The antibacterial properties of LBL-functionalized PAN nanofibrous membranes were evaluated using the Gram-positive bacterium Staphylococcus aureus and the Gram-negative Escherichia coli. Furthermore, the dependence of the antibacterial activity and anti-biofouling performance on the number of layers in the LBL films was investigated quantitatively. It was found that these LBL-modified nanofibrous membranes possessed high antibacterial activities, easy-cleaning properties and stability under physiological conditions, thus qualifying them as candidates for anti-biofouling coatings.