Anti-capsular activity of CuO nanoparticles against Acinetobacter baumannii produce efflux pump.

Copper oxide nanoparticles are modern kinds of antimicrobials, which may get a lot of interest in the clinical application. This study aimed to detect the anti-capsular activity of CuO nanoparticles against Acinetobacter baumannii produce efflux pump. Thirty-four different clinical A. baumannii isolates were collected and identified by the phenotypic and genetic methods by the recA gene as housekeeping. Antibiotic sensitivity and biofilm-forming ability, capsular formation were carried out. The effect of CuO nanoparticles on capsular isolates was detected, the synergistic effects of a combination CuO nanoparticles and gentamicin against A. baumannii were determined by micro broth checkerboard method, and the effect of CuO nanoparticles on the expression of ptk, espA and mexX genes was analyzed. Results demonstrated that CuO nanoparticles with gentamicin revealed a synergistic effect. Gene expression results show reducing the expression of these capsular genes by CuO nanoparticles is major conduct over reducing A. baumannii capsular action. Furthermore, results proved that there was a relationship between the capsule-forming ability and the absence of biofilm-forming ability. As bacterial isolates which were negative biofilm formation were positive in capsule formation and vice versa. In conclusion, CuO nanoparticles have the potential to be used as an anti-capsular agent against A. baumannii, and their combination with gentamicin can enhance their antimicrobial effect. The study also suggests that the absence of biofilm formation may be associated with the presence of capsule formation in A. baumannii. These findings provide a basis for further research on the use of CuO nanoparticles as a novel antimicrobial agent against A. baumannii and other bacterial pathogens, also to investigate the potential of CuO nanoparticles to inhibit the production of efflux pumps in A. baumannii, which are a major mechanism of antibiotic resistance.

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.

Hospital sink traps as a potential source of the emerging multidrug-resistant pathogen Cupriavidus pauculus: characterization and draft genome sequence of strain MF1.

Introduction. Cupriavidus pauculus is historically found in soil and water but has more recently been reported to cause human infection and death. Hospital sink traps can serve as a niche for bacterial persistence and a platform for horizontal gene transfer, with evidence of dissemination of pathogens in hospital plumbing systems driving nosocomial infection.Gap Statement. This paper presents the first C. pauculus strain isolated from a hospital sink trap. There are only six genome assemblies available on NCBI for C. pauculus; two of these are PacBio/Illumina hybrids. This paper presents the first ONT/Illumina hybrid assembly, with five contigs. The other assemblies available consist of 37, 38, 111 and 227 contigs. This paper also presents data on biofilm formation and lethal dose in Galleria mellonella; there is little published information describing these aspects of virulence.Aim. The aims were to identify the isolate found in a hospital sink trap, characterize its genome, and assess whether it could pose a risk to human health.Methodology. The genome was sequenced, and a hybrid assembly of short and long reads produced. Antimicrobial susceptibility was determined by the broth microdilution method. Virulence was assessed by measuring in vitro biofilm formation compared to Pseudomonas aeruginosa and in vivo lethality in Galleria mellonella larvae.Results. The isolate was confirmed to be a strain of C. pauculus, with a 6.8 Mb genome consisting of 6468 coding sequences and an overall G+C content of 63.9 mol%. The genome was found to contain 12 antibiotic resistance genes, 8 virulence factor genes and 33 metal resistance genes. The isolate can be categorized as resistant to meropenem, amoxicillin, amikacin, gentamicin and colistin, but susceptible to cefotaxime, cefepime, imipenem and ciprofloxacin. Clear biofilm formation was seen in all conditions over 72 h and exceeded that of P. aeruginosa when measured at 37 °C in R2A broth. Lethality in G. mellonella larvae over 48 h was relatively low.Conclusion. The appearance of a multidrug-resistant strain of C. pauculus in a known pathogen reservoir within a clinical setting should be considered concerning. Further work should be completed to compare biofilm formation and in vivo virulence between clinical and environmental strains, to determine how easily environmental strains may establish human infection. Infection control teams and clinicians should be aware of the emerging nature of this pathogen and further work is needed to minimize the impact of contaminated hospital plumbing systems on patient outcomes.

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.

Prevalence of Genes Involved in Colistin Resistance in Acinetobacter baumannii: First Report from Iraq.

Background and Aim: Colistin is increasingly being used as a "last-line" therapy to treat infections caused by multidrug-resistant (MDR) Acinetobacter baumannii isolates, when essentially no other options are available in these days. The aim of this study was to detect genes associated with colistin resistance in A. baumannii. Methods: One hundred twenty-one isolates of A. baumannii were collected from clinical and environmental samples during 2016 to 2018 in Baghdad. Isolates were diagnosed as A. baumannii by using morphological tests, Vitek-2 system, 16SrRNA PCR amplification, and sequencing. Antibiotic susceptibility test was carried out using disk diffusion method. Phenotypic detection of colistin resistance was performed by CHROMagar™ COL-APSE medium and broth microdilution method for the determination of the minimal inhibitory concentration. Molecular detection of genes responsible for colistin resistance in A. baumannii was performed by PCR. Results: Ninety-two (76%) of the 121 A. baumannii isolates were colistin resistant. Twenty-six (21.5%) of the 121 isolates showed positive growth on CHROMagar Acinetobacter base for MDR. PCR detected mcr-1, mcr-2, and mcr-3 genes in 89 (73.5%), 78 (64.5%), and 82 (67.8%) A. baumannii isolates, respectively. Seventy-eight (64.5%) of the 121 isolates harbored the integron intI2 gene and 81 (66.9%) contained intI3 gene. Moreover, 60 (49.6%) of the 121 isolates were positive for the quorum sensing lasI gene. Conclusion: The presence of a large percentage of colistin-resistant A. baumannii strains in Baghdad may be due to the presence of mobile genetic elements, and it is urgent to avoid unnecessary clinical use of colistin.

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.

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.

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.

The use of acetone to enhance the infiltration of HA nanoparticles into a demineralized dentin collagen matrix.

OBJECTIVES: This study investigates the role of acetone, as a carrier for nano-hydroxyapatite (nano-HA) in solution, to enhance the infiltration of fully demineralized dentin with HA nanoparticles (NPs). METHODS: Dentin specimens were fully demineralized and subsequently infiltrated with two types of water-based nano-HA solutions (one containing acetone and one without). Characterization of the dentin surfaces and nano-HA particles was performed using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The surface wettability and infiltration capacity of the nano-HA solutions were quantified by means of contact angle measurements and energy dispersive X-ray spectroscopy (EDS), respectively. Contact angle measurements were taken at baseline and repeated at regular intervals to assess the effect of acetone. The P and Ca levels of infiltrated dentin specimens were measured and compared to sound dentin and non-infiltrated controls. RESULTS: The presence of acetone resulted in an eight-fold decrease in the contact angles of the nano-HA solutions recorded on the surface of demineralized dentin compared to nano-HA solutions without acetone (one-way ANOVA, p<0.05). Perfect wetting of the demineralized dentin surface was achieved 5min after the application of the nano-HA solution containing acetone. Infiltration of demineralized dentin with the nano-HA solution containing acetone restored the lost mineral content by 50%, whereas the mean mineralization values for P and Ca in dentin treated with the acetone-free nano-HA solution were less than 6%. SIGNIFICANCE: Acetone was shown to act as a vehicle to enhance the capacity to infiltrate demineralized dentin with HA NPs. The successful infiltration of dentin collagen with HA NPs provides a suitable scaffold, whereby the infiltrated HA NPs have the potential to act as seeds that may initiate heterogenous mineral growth when exposed to an appropriate mineral-rich environment.

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.

Remineralization potential of fully demineralized dentin infiltrated with silica and hydroxyapatite nanoparticles.

OBJECTIVE: This study investigates the potential of a novel guided tissue regeneration strategy, using fully demineralized dentin infiltrated with silica and hydroxyapatite (HA) nanoparticles (NPs), to remineralize dentin collagen that is completely devoid of native hydroxyapatite. METHODS: Dentin blocks were fully demineralized with 4N formic acid and subsequently infiltrated with silica and HA NPs. The remineralizing potential of infiltrated dentin was assessed following a twelve week exposure to an artificial saliva solution by means of TEM, EDS and micro-CT. Measurements were taken at baseline and repeated at regular intervals for the duration of the study to quantify the P and Ca levels, the mineral volume percentage and mineral separation of the infiltrated dentin specimens compared to sound dentin and non-infiltrated controls. RESULTS: Infiltration of demineralized dentin with nano-HA restored up to 55% of the P and Ca levels at baseline. A local increase in the concentration of calcium phosphate compounds over a period of twelve weeks resulted in a higher concentration in P and Ca levels within the infiltrated specimens when compared to the non-infiltrated controls. Remineralization of demineralized dentin with silica NPs by immersion in artificial saliva was the most effective strategy, restoring 20% of the P levels of sound dentin. Micro-CT data showed a 16% recovery of the mineral volume in dentin infiltrated with silica NPs and a significant decrease in the mineral separation to levels comparable to sound dentin. SIGNIFICANCE: Demineralized dentin infiltrated with silica NPs appears to encourage heterogeneous mineralization of the dentin collagen matrix following exposure to an artificial saliva solution.

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.

Infiltration of demineralized dentin with silica and hydroxyapatite nanoparticles.

OBJECTIVES: The management of demineralized dentin resulting from dental caries or acid erosion remains an oral healthcare clinical challenge. This paper investigates, through a range of studies, the ability of colloidal silica and hydroxyapatite (HA) nanoparticles to infiltrate the collagen structure of demineralized dentin. METHODS: Dentin samples were completely demineralized in 4 N formic acid. The remaining collagen matrix of the dentin samples was subsequently infiltrated with a range of nano-particulate colloidal silica and HA solutions. The effectiveness and extent of the infiltration was evaluated by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and energy dispersive X-ray spectroscopy (EDS). RESULTS: Silica nanoparticles have the ability to penetrate dentin and remain embedded within the collagen matrix. It is suggested that particle size plays a major role in the degree of dentin infiltration, with smaller diameter particles demonstrating a greater infiltrative capacity. The infiltration of demineralized dentin with sol-gel HA nanoparticles was limited but was significantly increased when combined with the deflocculating agent sodium hexametaphosphate. The use of acetone as a transport vehicle is reported to enhance the infiltration capacity of sol-gel HA nanoparticles. SIGNIFICANCE: Collagen infiltrated with HA and silica nanoparticles may provide a suitable scaffold for the remineralization of dentin, whereby the infiltrated particles act as seeds within the collage matrix and given the appropriate remineralizing environment, mineral growth may occur.