Surface PEGylation suppresses pulmonary effects of CuO in allergen-induced lung inflammation.

BACKGROUND: Copper oxide (CuO) nanomaterials are used in a wide range of industrial and commercial applications. These materials can be hazardous, especially if they are inhaled. As a result, the pulmonary effects of CuO nanomaterials have been studied in healthy subjects but limited knowledge exists today about their effects on lungs with allergic airway inflammation (AAI). The objective of this study was to investigate how pristine CuO modulates allergic lung inflammation and whether surface modifications can influence its reactivity. CuO and its carboxylated (CuO COOH), methylaminated (CuO NH3) and PEGylated (CuO PEG) derivatives were administered here on four consecutive days via oropharyngeal aspiration in a mouse model of AAI. Standard genome-wide gene expression profiling as well as conventional histopathological and immunological methods were used to investigate the modulatory effects of the nanomaterials on both healthy and compromised immune system. RESULTS: Our data demonstrates that although CuO materials did not considerably influence hallmarks of allergic airway inflammation, the materials exacerbated the existing lung inflammation by eliciting dramatic pulmonary neutrophilia. Transcriptomic analysis showed that CuO, CuO COOH and CuO NH3 commonly enriched neutrophil-related biological processes, especially in healthy mice. In sharp contrast, CuO PEG had a significantly lower potential in triggering changes in lungs of healthy and allergic mice revealing that surface PEGylation suppresses the effects triggered by the pristine material. CONCLUSIONS: CuO as well as its functionalized forms worsen allergic airway inflammation by causing neutrophilia in the lungs, however, our results also show that surface PEGylation can be a promising approach for inhibiting the effects of pristine CuO. Our study provides information for health and safety assessment of modified CuO materials, and it can be useful in the development of nanomedical applications.

Review of Antimicrobial Nanocoatings in Medicine and Dentistry: Mechanisms of Action, Biocompatibility Performance, Safety, and Benefits Compared to Antibiotics.

This review discusses topics relevant to the development of antimicrobial nanocoatings and nanoscale surface modifications for medical and dental applications. Nanomaterials have unique properties compared to their micro- and macro-scale counterparts and can be used to reduce or inhibit bacterial growth, surface colonization and biofilm development. Generally, nanocoatings exert their antimicrobial effects through biochemical reactions, production of reactive oxygen species or ionic release, while modified nanotopographies create a physically hostile surface for bacteria, killing cells via biomechanical damage. Nanocoatings may consist of metal nanoparticles including silver, copper, gold, zinc, titanium, and aluminum, while nonmetallic compounds used in nanocoatings may be carbon-based in the form of graphene or carbon nanotubes, or composed of silica or chitosan. Surface nanotopography can be modified by the inclusion of nanoprotrusions or black silicon. Two or more nanomaterials can be combined to form nanocomposites with distinct chemical or physical characteristics, allowing combination of different properties such as antimicrobial activity, biocompatibility, strength, and durability. Despite their wide range of applications in medical engineering, questions have been raised regarding potential toxicity and hazards. Current legal frameworks do not effectively regulate antimicrobial nanocoatings in matters of safety, with open questions remaining about risk analysis and occupational exposure limits not considering coating-based approaches. Bacterial resistance to nanomaterials is also a concern, especially where it may affect wider antimicrobial resistance. Nanocoatings have excellent potential for future use, but safe development of antimicrobials requires careful consideration of the "One Health" agenda, appropriate legislation, and risk assessment.

From microbes to ecosystems: a review of the ecological effects of biodegradable plastics.

Biodegradable plastics have been proposed as a potential solution to plastic pollution, as they can be biodegraded into their elemental components by microbial action. However, the degradation rate of biodegradable plastics is highly variable across environments, leading to the potential for accumulation of plastic particles, chemical co-contaminants and/or degradation products. This paper reviews the toxicological effects of biodegradable plastics on species and ecosystems, and contextualises these impacts with those previously reported for conventional polymers. While the impacts of biodegradable plastics and their co-contaminants across levels of biological organisation are poorly researched compared with conventional plastics, evidence suggests that individual-level effects could be broadly similar. Where differences in the associated toxicity may arise is due to the chemical structure of biodegradable polymers which should facilitate enzymatic depolymerisation and the utilisation of the polymer carbon by the microbial community. The input of carbon can alter microbial composition, causing an enrichment of carbon-degrading bacteria and fungi, which can have wider implications for carbon and nitrogen dynamics. Furthermore, there is the potential for toxic degradation products to form during biodegradation, however understanding the environmental concentration and effects of degradation products are lacking. As global production of biodegradable polymers continues to increase, further evaluation of their ecotoxicological effects on organisms and ecosystem function are required.

The biocompatibility of silver and nanohydroxyapatite coatings on titanium dental implants with human primary osteoblast cells.

Silver nanoparticles (Ag NPs) are antimicrobial, with potential uses in medical implants, but Ag NPs alone can also be toxic to mammalian cells. This study aimed to enhance the biocompatibility of Ag NP-coated titanium dental implants with hydroxyapatite (HA) applied to the surface. Ti6Al4V discs were coated with Ag NPs, Ag NPs plus HA nanoparticles (Ag + nHA), or Ag NPs plus HA microparticles (Ag + mHA). The stability of coatings was explored and the biocompatibility with primary human osteoblasts over 7 days. Results showed that Ti6Al4V discs were successfully coated with silver and HA. The primary particle size of nHA and mHA were 23.90 ±â€¯1.49 nm and 4.72 ±â€¯0.38 µm respectively. Metal analysis showed that underlying silver coatings remain stable in DMEM culture media, but the presence of FBS in the media caused some initial (clinically beneficial) release of dissolved silver. With additions of HA, osteoblasts were adherent, had normal morphology, negligible lactate dehydrogenase (LDH) leak, and showed alkaline phosphatase (ALP) activity. Cell viability was around 70% throughout the Ag + nHA treatment. Overall, the implants coated with Ag + nHA maintained a higher degree of biocompatibility compared to those coated with Ag + mHA, or Ag NPs alone, suggesting the former has a benefit for clinical use.

Antibacterial properties of silver nanoparticles grown in situ and anchored to titanium dioxide nanotubes on titanium implant against Staphylococcus aureus.

Medical grade titanium alloy, Ti-6Al-4V, with TiO2 nanotubes (TiO2-NTs) grown on the surface and then decorated with silver nanoparticles (Ag NPs) is proposed to enhance the antimicrobial properties of the bone/dental implants. However, the decoration with Ag NPs is not consistent and there are concerns about the direct contact of Ag NPs with human tissue. The aim of this study was to achieve a more even coverage of Ag NPs on TiO2-NTs and determine their biocidal properties against Staphylococcus aureus, with and without a top coat of nano hydroxyapatite (nHA). The decoration with Ag NPs was optimised by adjusting the incubation time of the TiO2-NTs in a silver ammonia solution, and using biocompatible δ-gluconolactone as a reducing agent. The optimum incubation in silver ammonia was 7 min, and resulted in evenly distributed Ag NPs with an average diameter of 47.5 ± 1.7 nm attached to the surface of the nanotubes. The addition of nHA did not compromise the antimicrobial properties of the materials; high-resolution electron microscopy showed S. aureus did not grow on the composite with nHA and with >80% biocidal activity measured by the LIVE/DEAD assay, also limited lactate production. Dialysis experiment confirmed the stability of the coatings, and showed a slow release of dissolved silver (3.27 ± 0.15 µg/L over 24 h) through the top coat of nHA.

Carbon Nanotube Reinforced Hydroxyapatite Nanocomposites As Bone Implants: Nanostructure, Mechanical Strength And Biocompatibility.

PURPOSE: Hydroxyapatite (HA) is a biologically active ceramic which promotes bone growth, but it suffers from relatively weak mechanical properties. Multi-walled carbon nanotubes (MWCNTs) have high tensile strength and a degree of stiffness that can be used to strengthen HA; potentially improving the clinical utility of the bone implant. METHODS: HA was precipitated by the wet precipitation method in the presence of pristine (p) or functionalised (f) MWCNTs, and polyvinyl alcohol (PVA) or hexadecyl trimethyl ammonium bromide (HTAB) as the surfactant. The resulting composites were characterised and the diametral tensile strength and compressive strength of the composites were measured. To determine the biocompatibility of the composites, human osteoblast cells (HOB) were proliferated in the presence of the composites for 7 days. RESULTS: The study revealed that both the MWCNTs and surfactants play a crucial role in the nucleation and growth of the HA. Composites made with f-MWCNTs were found to have better dispersion and better interaction with the HA particles compared to composites with p-MWCNTs. The mechanical strength was improved in all the composites compared to pure HA composites. The biocompatibility study showed minimal LDH activity in the media confirming that the composites were biocompatible. Similarly, the ALP activity confirmed that the cells grown on the composites containing HTAB were comparable to the control whereas the composites containing PVA surfactant showed significantly reduced ALP activity. CONCLUSIONS: The study shows that the composites made of f-MWCNTs HTAB are stronger than pure HA composites and biocompatible making it a suitable material to study further.

Multilayered composite coatings of titanium dioxide nanotubes decorated with zinc oxide and hydroxyapatite nanoparticles: controlled release of Zn and antimicrobial properties against Staphylococcus aureus.

Purpose: This study aimed to decorate the surface of TiO2 nanotubes (TiO2 NTs) grown on medical grade Ti-6Al-4V alloy with an antimicrobial layer of nano zinc oxide particles (nZnO) and then determine if the antimicrobial properties were maintained with a final layer of nano-hydroxyapatite (HA) on the composite. Methods: The additions of nZnO were attempted at three different annealing temperatures: 350, 450 and 550 °C. Of these temperatures, 350°C provided the most uniform and nanoporous coating and was selected for antimicrobial testing. Results: The LIVE/DEAD assay showed that ZnCl2 and nZnO alone were >90% biocidal to the attached bacteria, and nZnO as a coating on the nanotubes resulted in around 70% biocidal activity. The lactate production assay agreed with the LIVE/DEAD assay. The concentrations of lactate produced by the attached bacteria on the surface of nZnO-coated TiO2 NTs and ZnO/HA-coated TiO2 NTs were 0.13±0.03 mM and 0.37±0.1 mM, respectively, which was significantly lower than that produced by the bacteria on TiO2 NTs alone, 1.09±0.30 mM (Kruskal-Wallis, P<0.05, n=6). These biochemical measurements were correlated with electron micrographs of cell morphology and cell coverage on the coatings. Conclusion: nZnO on TiO2 NTs was a stable and antimicrobial coating, and most of the biocidal properties remained in the presence of nano-HA on the coating.

Antifungal properties and biocompatibility of silver nanoparticle coatings on silicone maxillofacial prostheses in vitro.

Patients with facial prostheses suffer from yeast, Candida albicans, infections. This study aimed to determine the biocompatibility and antifungal properties of silicone facial prostheses coated with silver nanoparticles (Ag NPs) in vitro. Medical grade silicone discs were coated with 5 and 50 mg L-1 dispersions of either Ag NPs or AgNO3 . Coatings were fully characterized using scanning electron microscopy and energy dispersive X-ray spectroscopy. The biocompatibility was examined using human dermal fibroblasts (Hs68), whereas antifungal efficacy was tested against C. albicans (NCPF-3179). The fibroblast viability was assessed by measuring lactate dehydrogenase (LDH) activity, protein content and tissue electrolytes. There were no effects on the LDH activity of fibroblast cell homogenates, and leak of LDH activity into external media remained low (0.1-0.2 IU mL-1 ). Sublethal effects of Ag NP coatings on membrane permeability/ion balance was not observed, as measured by stable homogenate Na+ and K+ concentrations. Some Ag (13 mg L-1 ) was detected from the AgNO3 coatings in the media, but total Ag remained below detection limit (<1.2 µg L-1 ) for the Ag NP coatings; indicating the latter were stable. When fibroblasts grown on silver coatings were challenged with C. albicans, the Ag NP coating was effective at preventing fungal growth as measured by ethanol production by the yeast, and without damaging the fibroblasts. Ethanol production decreased from 43.2 ± 25.02 in controls to 3.6 µmol mL-1 in all the silver treatments. Data shows that silicone prosthetic materials coated with Ag NPs are biocompatible with fibroblast cells in vitro and show antifungal properties. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 1038-1051, 2018.

Anodised TiO2 nanotubes as a scaffold for antibacterial silver nanoparticles on titanium implants.

Medical grade titanium alloy is widely used for bone/dental implants, but the material alone has no innate antimicrobial properties that would reduce infection risk following surgery. However, silver nanoparticles (Ag NPs) are known to be antibacterial. This study investigated the growth of Ag NPs on titanium dioxide nanotubes (TiO2 NTs) on Ti-6Al-4V discs. The TiO2 NTs were grown on the Ti alloy using an electrochemical method, and then decorated with Ag NPs. The Ag NPs were synthesised by chemical reduction using δ-gluconolactone. A silver ammonia solution (silver nitrate + liquid ammonia) was used as the source of silver. Two separate approaches were used: (1) The δ-gluconolactone was mixed with the silver ammonia and then exposed to the TiO2 NTs (the 'mixing method'), which produced micron-sized clusters of the Ag NPs. (2) The TiO2 NTs were exposed to the silver ammonia first and then to δ-gluconolactone (the 'sequential addition method'), which resulted in the formation of nano-sized clusters of the nanoparticles. The Ag-TiO2 composites were confirmed by scanning electron microscopy and the elements analysed using energy dispersive X-ray spectroscopy (EDS). The composite coatings were exposed to a simulated body fluid for 24 h in order to determine the total Ag released. The release from the micron-sized clusters from the mixing method (14.6 ±â€¯0.67 ppm) was higher than that from the nano-sized clusters (4.05 ±â€¯0.36 ppm) when 0.015 M of silver ammonia was used. Additionally, Staphylococcus aureus, was cultured on the composite coatings for 24 h. Both the micron- and nano-sized clusters of the Ag NPs were found to be antibacterial using the Live/Dead assay. Overall, δ-gluconolactone was successfully used to reduce silver to Ag NPs on the surface of TiO2 NTs. The sequential addition method was the preferred method of synthesis because of its slower silver release, better coverage of the Ag-NPs on the TiO2 NTs and strong antibacterial properties.

Review of nanomaterials in dentistry: interactions with the oral microenvironment, clinical applications, hazards, and benefits.

Interest in the use of engineered nanomaterials (ENMs) as either nanomedicines or dental materials/devices in clinical dentistry is growing. This review aims to detail the ultrafine structure, chemical composition, and reactivity of dental tissues in the context of interactions with ENMs, including the saliva, pellicle layer, and oral biofilm; then describes the applications of ENMs in dentistry in context with beneficial clinical outcomes versus potential risks. The flow rate and quality of saliva are likely to influence the behavior of ENMs in the oral cavity, but how the protein corona formed on the ENMs will alter bioavailability, or interact with the structure and proteins of the pellicle layer, as well as microbes in the biofilm, remains unclear. The tooth enamel is a dense crystalline structure that is likely to act as a barrier to ENM penetration, but underlying dentinal tubules are not. Consequently, ENMs may be used to strengthen dentine or regenerate pulp tissue. ENMs have dental applications as antibacterials for infection control, as nanofillers to improve the mechanical and bioactive properties of restoration materials, and as novel coatings on dental implants. Dentifrices and some related personal care products are already available for oral health applications. Overall, the clinical benefits generally outweigh the hazards of using ENMs in the oral cavity, and the latter should not prevent the responsible innovation of nanotechnology in dentistry. However, the clinical safety regulations for dental materials have not been specifically updated for ENMs, and some guidance on occupational health for practitioners is also needed. Knowledge gaps for future research include the formation of protein corona in the oral cavity, ENM diffusion through clinically relevant biofilms, and mechanistic investigations on how ENMs strengthen the tooth structure.

The antibacterial effects of silver, titanium dioxide and silica dioxide nanoparticles compared to the dental disinfectant chlorhexidine on Streptococcus mutans using a suite of bioassays.

Metal-containing nanomaterials have the potential to be used in dentistry for infection control, but little is known about their antibacterial properties. This study investigated the toxicity of silver (Ag), titanium dioxide and silica nanoparticles (NPs) against the oral pathogenic species of Streptococcus mutans, compared to the routine disinfectant, chlorhexidine. The bacteria were assessed using the minimum inhibitory concentration assay for growth, fluorescent staining for live/dead cells, and measurements of lactate. All the assays showed that Ag NPs had the strongest antibacterial activity of the NPs tested, with bacterial growth also being 25-fold lower than that in chlorhexidine. The survival rate of bacteria under the effect of 100 mg l(-1) Ag NPs in the media was 2% compared to 60% with chlorhexidine, while the lactate concentration was 0.6 and 4.0 mM, respectively. Silica and titanium dioxide NPs had limited effects. Dialysis experiments showed negligible silver dissolution. Overall, Ag NPs were the best disinfectant and performed better than chlorhexidine. Improvements to the MIC assay are suggested.

Inhibition of biofilm formation and antibacterial properties of a silver nano-coating on human dentine.

The survival of pathogenic bacteria in the oral cavity depends on their successful adhesion to dental surfaces and their ability to develop into biofilms, known as dental plaque. Bacteria from the dental plaque are responsible for the development of dental caries, gingivitis, periodontitis, stomatitis and peri-implantitis. Certain metal nanoparticles have been suggested for infection control and the management of the oral biofilm. Here, it is shown that application of a silver nano-coating directly on dentine can successfully prevent the biofilm formation on dentine surfaces as well as inhibit bacterial growth in the surrounding media. This silver nano-coating was found to be stable (>98.8%) and to maintain its integrity in biological fluids. Its antibacterial activity was compared to silver nitrate and the widely used clinical antiseptic, chlorhexidine. The bacterial growth and cell viability were quantitatively assessed by measuring the turbidity, proportion of live and dead cells and lactate production. All three bioassays showed that silver nanoparticles and silver nitrate dentine coatings were equally highly bactericidal (>99.5%), while inhibiting bacterial adhesion. However, the latter caused significant dentine discolouration (ΔE* = 50.3). The chlorhexidine coating showed no antibacterial effect. Thus, silver nanoparticles may be a viable alternative to both chlorhexidine and silver nitrate, protecting from dental plaque and secondary caries when applied as a dentine coating, while they may provide the platform for creating anti-biofilm surfaces in medical devices and other biomedical applications.

A simplified method for determining titanium from TiO2 nanoparticles in fish tissue with a concomitant multi-element analysis.

The reliable detection of nanoparticles (NPs) in fish tissue is required to support ecotoxicological research and food safety investigations. Therefore the current work aimed to develop a simple method to determine Ti from TiO2 NPs in fish tissue whilst simultaneously measuring other elements in the sample. Spike recovery tests showed no differences when digestion was conducted in glass or plastic vials, there was stirring or sonication of the samples, or when sodium dodecyl sulfate was added. However, the addition of 2% Triton X-100 and sonicating and then vortexing of samples immediately prior to analysis did improve recovery (approximately 20% to >90% in trout gill and muscle samples). Method precision and accuracy were good with coefficients of variation <7%. Copper spike recovery results showed that the method is also suitable for multi-element analysis in the same samples. This improved method is simple with high throughput and represents a marked improvement for routine determination Ti from TiO2 NPs in fish tissues.