The Push-Out Bond Strength, Surface Roughness, and Antimicrobial Properties of Endodontic Bioceramic Sealers Supplemented with Silver Nanoparticles.

This study evaluated push-out bond test (POBT), surface roughness, and antimicrobial properties against Enterococcus faecalis of bioceramic sealers supplemented with silver nanoparticles (AgNPs). The sealers tested were CeraSeal®, EndoSequence® BC SealerTM, and Bio-C® Sealer. The POBT was measured with a Universal Testing Machine, and the type of failure was evaluated with a stereomicroscope. The roughness average (Sa) and peak-valley height (Sy) values were evaluated by atomic force microscopy. The bacterial growth inhibition was evaluated using a disk diffusion test, and antimicrobial activity was determined with the plate microdilution method. The POBT showed no significant difference between sealers with and those without NPs in cervical and apical thirds (p > 0.05). In the middle third, the adhesion force was significant for Endosequence BC Sealer® (p < 0.05). The results showed that the Sa and Sy parameters, when AgNPs were added, did not show a statistically significant difference compared to the groups without nanoparticles (p > 0.05). All tested sealers showed bacterial growth inhibition, but no significant difference was found. Their efficacy, in descending order of antibacterial activity when AgNPs were added, is as follows: EndoSequence® BC SealerTM > Bio-C® Sealer > CeraSeal®. The incorporation of AgNPs into bioceramics improves antimicrobial activity without affecting mechanical properties.

Proteomic analysis of an Enterococcus faecalis mutant generated against the exposure to silver nanoparticles.

INTRODUCTION: Nanoparticles (NPs) have been widely studied as an alternative to antibiotic use due to their antimicrobial properties at lower concentrations. Enterococcus faecalis is a facultative Gram-positive microorganism inhabiting the gastrointestinal tract of humans and animals. It can also be present in other environments such as the oral cavity, water, sewage, soil and food. AIMS: We evaluated whether E. faecalis could develop resistance to silver NPs (AgNPs) after exposure to sublethal concentrations of the NPs. METHODS AND RESULTS: Proteomic analyses revealed that different pathways were activated during the acquired resistance under sublethal concentrations, and selected genes were validated by qPCR. CONCLUSIONS: The results of this study showed that E. faecalis is capable of generating resistance to AgNPs. SIGNIFICANCE AND IMPACT OF THE STUDY: To avoid the generation of resistance against AgNPs, future use of these NPs should be combined with other NPs prepared with different metals to prevent the dissemination of resistant strains.

Reusability in visible light of titanate nanotubes for the removal of organic pollutants: role of calcination temperature.

Titanate nanotubes (NTs) were synthesised by the hydrothermal method and later calcined at temperatures between 100-500°C. The calcined NTs were characterised and evaluated in the physicochemical adsorption of the safranin dye and photocatalytic degradation of caffeine. The materials calcined at low temperatures displayed a tubular structure and the H2Ti3O7 crystalline phase, which was transformed into anatase nanoparticles at 400°C. The NTs treated at 100°C showed the highest adsorption capacity (94%). Safranin was adsorbed through an ion-exchange mechanism, following the Langmuir isotherm and a pseudo-second-order kinetic model. While NTs calcined at lower temperatures were better for adsorption, the photocatalytic degradation of caffeine increased in samples calcined at higher temperatures with a maximum removal of 72%. The photocatalytic behaviour of the NT samples confirmed that the crystalline anatase structure in conjunction with structural OH groups enhanced the photocatalytic activity. The addition of isopropanol as a scavenger confirmed the important role played by the •OH radicals in the photocatalytic process. NTs calcined at 300°C were efficient for both adsorption and photocatalytic processes. Due to its efficiency, this sample was reused after dye adsorption for the photocatalytic degradation of caffeine under visible light due to its enhanced absorbance in the visible region. This research work shows the potential of NTs for wastewater purification.

Effect of surface characteristics on the antibacterial properties of titanium dioxide nanotubes produced in aqueous electrolytes with carboxymethyl cellulose.

Nanotubular structures were produced on a commercially pure titanium surface by anodization in an aqueous electrolyte that contained carboxymethyl cellulose and sodium fluoride. The internal diameters obtained were about 100, 48, and 9.5 nm, respectively. Several heat treatments at 200, 350, and 600°C were made to produce nanotubes with different titanium dioxide polymorphs (anatase, rutile). All tested surfaces were superhydrophilic, this behavior was maintained after at least 30 days, regardless of the heat treatment. Although in previous works the nanotube features effect on the bacteria behavior had been studied; this item still unclear. For the best of our knowledge, the effect of small internal diameters (about 10 nm) with and without heat treatment and with and without ultraviolet (UV) irradiation on the bacteria strains comportment has not been reported. From our results, both the internal diameter and the postanodized treatments have an effect on the bacteria strains comportment. All nanotubular coatings UV treated and heat treated at 350 and 600°C; despite they have different inner diameters, inhibit the bacteria growth of both Staphylococcus aureus and Pseudomonas aeruginosa strains. The nanotubular coatings obtained at 20 V and heat treated at 350°C produced the lower bacteria adhesion against both strains evaluated.

Atmospheric Corrosion, Antibacterial Properties, and Toxicity of Silver Nanoparticles Synthesized by Two Different Routes.

Silver nanoparticles (AgNPs) have been widely employed or incorporated into different materials in biological application, due to their antibacterial properties. Therefore, antimicrobial capacity and cytotoxicity have been highly studied. However, most of these reports do not consider the possible corrosion of the nanomaterials during their exposure to atmospheric conditions since AgNPs undergo a transformation when they come in contact with a particular environment. Derived from this, the functionality and properties of the nanoparticles could decrease noticeably. The most common silver corrosion process occurs by the interaction of AgNPs with sulfur species (H2S) present in the atmospheric air, forming a corrosion layer of silver sulfide around the AgNPs, thus inhibiting the release of the ions responsible for the antimicrobial activity. In this work, AgNPs were synthesized using two different methods: one of them was based on a plant extract (Brickellia cavanillesii), and the other one is the well-known method using sodium borohydride (NaBH4). Chemical stability, corrosion, antibacterial activity, and toxic activity were evaluated for both sets of prepared samples, before and after exposition to atmospheric air for three months. The structural characterization of the samples, in terms of crystallinity, chemical composition, and morphology, evidenced the formation of link structures with nanobridges of Ag2S for non- "green" AgNPs after the air exposition and the intact preservation of silver core for the "green" sample. The antibacterial activity showed a clear improvement in the antimicrobial properties of silver in relation to the "green" functionalization, particle size control, and size reduction, as well as the preservation of the properties after air exposition by the effective "green" protection. The cytotoxicity effect of the different AgNPs against mononuclear cells showed a notable increment in the cell viability by the "green" functionalization.

Therapeutic Use of Silver Nanoparticles in the Prevention and Arrest of Dental Caries.

Dental caries is one of the major diseases of the oral cavity affecting humans worldwide. Different alternatives have been used for its control, but its incidence and prevalence are still high. On the other hand, silver has been used for centuries due to its antimicrobial properties. With advances in nanotechnology, the use and research in nanomaterials has increased, recently, and silver nanoparticles have become an essential part of the dental practice, giving materials physical and chemical improvements in their properties, used for their antibacterial capacity preventing and arresting dental caries. The objective of this review was to examine the use of silver nanoparticles, in the treatment of dental caries in the remineralization of teeth hard tissues, as well as the antimicrobial potential, cytotoxicity, and long-term effectiveness.

Effective control of biofilms by photothermal therapy using a gold nanorod hydrogel.

Biofilms are matrices synthesized by bacteria containing polysaccharides, DNA, and proteins. The development of biofilms in infectious processes can induce a chronic inflammatory response that may progress to the destruction of tissues. The treatment of biofilms is difficult because they serve as a bacterial mechanism of defense and high doses of antibiotics are necessary to treat these infections with limited positive results. It has been demonstrated that photothermal therapy using gold nanorods (AuNRs) is an attractive treatment because of its anti-biofilm activity. The purpose of this work was to generate a novel chitosan-based hydrogel embedded with AuNRs to evaluate its anti-biofilm activity. AuNRs were synthesized by the seed-mediated growth method and mixed with the chitosan-based hydrogel. Hydrogels were characterized and tested against two bacterial strains by irradiating the produced biofilm in the presence of the nanoformulation with a laser adjusted at the near infrared spectrum. In addition, the safety of the nanoformulation was assessed with normal human gingival fibroblasts. Results showed that a significant bacterial killing was measured when biofilms were exposed to an increase of 10°C for a short time of 2 min. Moreover, no cytotoxicity was measured when normal gingival fibroblasts were exposed to the nanoformulation using the bactericidal conditions. The development of the reported formulation can be used as a direct application to treat periodontal diseases or biofilm-produced bacteria that colonize the oral cavity.

Molecular Mechanisms of Bacterial Resistance to Metal and Metal Oxide Nanoparticles.

The increase in bacterial resistance to one or several antibiotics has become a global health problem. Recently, nanomaterials have become a tool against multidrug-resistant bacteria. The metal and metal oxide nanoparticles are one of the most studied nanomaterials against multidrug-resistant bacteria. Several in vitro studies report that metal nanoparticles have antimicrobial properties against a broad spectrum of bacterial species. However, until recently, the bacterial resistance mechanisms to the bactericidal action of the nanoparticles had not been investigated. Some of the recently reported resistance mechanisms include electrostatic repulsion, ion efflux pumps, expression of extracellular matrices, and the adaptation of biofilms and mutations. The objective of this review is to summarize the recent findings regarding the mechanisms used by bacteria to counteract the antimicrobial effects of nanoparticles.

Cytotoxic and Bactericidal Effect of Silver Nanoparticles Obtained by Green Synthesis Method Using Annona muricata Aqueous Extract and Functionalized with 5-Fluorouracil.

Nanomaterials obtained by green synthesis technologies have been widely studied in recent years owing to constitute cost-effective and environmental-friendly methods. In addition, there are several works that report the simultaneous performance of the reducer agent as a functionalizing agent, modifying the properties of the nanomaterial. As a simple and economical synthesis methodology, this work presents a method to synthesize silver nanoparticles (AgNPs) using Annona muricata aqueous extract and functionalized with 5-fluorouracil (5-FU). The processes of reduction, nucleation, and functionalization of the nanoparticles were analyzed by UV-Vis absorption spectroscopy, and it was found that they are the function of the contact time of the metal ions with the extract. The structural characterization was carried out by transmission electron microscopy (TEM) and X-ray diffraction patterns (XRD). The antibacterial properties of the synthetized nanomaterials were tested using minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against Enterococcus faecalis, Staphylococcus aureus, and Escherichia coli growth.

Antimicrobial Properties of Copper Nanoparticles and Amino Acid Chelated Copper Nanoparticles Produced by Using a Soya Extract.

This paper reports a comparison of the antibacterial properties of copper-amino acids chelates and copper nanoparticles against Escherichia coli, Staphylococcus aureus, and Enterococcus faecalis. These copper-amino acids chelates were synthesized by using a soybean aqueous extract and copper nanoparticles were produced using as a starting material the copper-amino acids chelates species. The antibacterial activity of the samples was evaluated by using the standard microdilution method (CLSI M100-S25 January 2015). In the antibacterial activity assays copper ions and copper-EDTA chelates were included as references, so that copper-amino acids chelates can be particularly suitable for acting as an antibacterial agent, so they are excellent candidates for specific applications. Additionally, to confirm the antimicrobial mechanism on bacterial cells, MTT assay (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) was carried out. A significant enhanced antimicrobial activity and a specific strain were found for copper chelates over E. faecalis. Its results would eventually lead to better utilization of copper-amino acids chelate for specific application where copper nanoparticles can be not used.

Effects of silver nanoparticles on the bonding of three adhesive systems to fluorotic enamel.

The objective was to evaluate the effect of adding silver nanoparticles into three commercial adhesive systems (Excite™, Adper Prompt L-Pop™ and AdheSE™). Nanoparticles were prepared by a chemical method then mixed with the commercial adhesive systems. This was later applied to the fluorotic enamel, and then micro-tensile bond strength, contact angle measurements and scanning electron microscopy observations were conducted. The commercial adhesive systems achieved the lowest micro-tensile bond strength (Excite™: 11.0±2.1, Adper Prompt L-Pop™: 14.0±5.4 and AdheSE™: 16.0±3.0 MPa) with the highest adhesive failure mode related with the highest contact angle (46.0±0.6º, 30.0±0.5º and 28.0±0.4º respectively). The bond strength achieved in all the experimental adhesive systems (19.0±5.4, 20.0±4.0 and 19.0±3.5 MPa respectively) was statistically higher (p<0.05) than the control and showed the highest cohesive failures related to the lowest contact angle. Adding silver nanoparticles in order to decrease the contact angle improve the adhesive system wetting and its bond strength.

Effect of Silver Nanoparticle-Added Pit and Fissure Sealant in the Prevention of Dental Caries in Children.

OBJECTIVE: The objective of this study was to evaluate the effects of pit and fissure sealant mixed with silver nanoparticles on dental caries, by means of monthly measurement of fluorescence with DIAGNOdent over six months. STUDY DESIGN: This study was divided in two phases: experimental and clinical. In the experimental phase, the adhesion and microleakage of the pit and fissure sealant experiment were evaluated. Two groups of 10 teeth, without serious carious lesions, were included. Conventional (group A) and silver nanoparticles (group B) were added to the pit and fissure sealant. For the clinical phase, a split-mouth study was performed on 40 children aged 6-10 years old with healthy, erupted permanent first molars. A conventional pit and fissure sealant or a silver nanoparticle-mixed sealant was randomly placed. Repeated measures analysis was performed. RESULTS: Conventional sealant presented an average microleakage of 30.6%, and the silver nanoparticle-mixed sealant showed 33.6% (P=NS). A three times greater reduction in fluorescence was found in the silver nanoparticles group compared to the conventional group (P<0.05). No sex- or age-based associations were found. CONCLUSIONS: The silver nanoparticle-mixed sealant reduced tooth demineralization significantly and likely increased remineralization, compared to the conventional sealant.

Facile Synthesis, Characterization, and Cytotoxic Activity of Europium-Doped Nanohydroxyapatite.

The objective of this study was to synthetize europium-doped nanohydroxyapatite using a simple aqueous precipitation method and, thereafter, characterize and impregnate selected samples with 5-fluorouracil in order to explore the properties and the releasing capacity of this material. The nanohydroxyapatite was doped with 3, 5, 10, and 20 wt% of europium. The obtained samples were characterized after they were dried at 80°C and hydrothermal treated at 120°C by 2 hours. The samples were analyzed by transmission electron microscopy, X-ray diffraction analysis, Fourier transform infrared spectroscopy, and photoluminescence. Also, impregnation and release of 5-fluorouracil were assessed in PBS. The toxicity effects of all samples were studied using viability assays on human fibroblasts cells (HGF-1) in vitro. The sizes of the crystallites were about 10-70 nm with irregular morphology and present the phase corresponding to the JCPDS card 9-0432 for hydroxyapatite. The results of the toxicity experiments indicated that doped and undoped powders are biocompatible with fibroblasts cells. Hydroxyapatite samples doped with 5% of europium and loaded with 5-fluorouracil release almost 7 mg/L of the drug after 60 minutes in PBS and decrease the viability of HeLa cells after 24 hours.

Antimicrobial Properties of Biofunctionalized Silver Nanoparticles on Clinical Isolates of Streptococcus mutans and Its Serotypes.

(1) Background: Streptococcus mutans (S. mutans) is the principal pathogen involved in the formation of dental caries. Other systemic diseases have also been associated with specific S. mutans serotypes (c, e, f, and k). Silver nanoparticles (SNP) have been demonstrated to have good antibacterial effects against S. mutans; therefore, limited studies have evaluated the antimicrobial activity of biofunctionalized SNP on S. mutans serotypes. The purpose of this work was to prepare and characterize coated SNP using two different organic components and to evaluate the antimicrobial activity of SNP in clinical isolates of S. mutans strains and serotypes; (2) Methods: SNP with bovine serum albumin (BSA) or chitosan (CS) coatings were prepared and the physical, chemical and microbiological properties of SNP were evaluated; (3) Results: Both types of coated SNP showed antimicrobial activity against S. mutans bacteria and serotypes. Better inhibition was associated with smaller particles and BSA coatings; however, no significant differences were found between the different serotypes, indicating a similar sensitivity to the coated SNP; (4) Conclusion: This study concludes that BSA and CS coated SNP had good antimicrobial activity against S. mutans strains and the four serotypes, and this study suggest the widespread use of SNP as an antimicrobial agent for the inhibition of S. mutans bacteria.

Anti-biofilm activity of silver nanoparticles against different microorganisms.

Biofilms confer protection from adverse environmental conditions and can be reservoirs for pathogenic organisms and sources of disease outbreaks, especially in medical devices. The goal of this research was to evaluate the anti-biofilm activities of silver nanoparticles (AgNPs) against several microorganisms of clinical interest. The antimicrobial activity of AgNPs was tested within biofilms generated under static conditions and also under high fluid shears conditions using a bioreactor. A 4-log reduction in the number of colony-forming units of Pseudomonas aeruginosa was recorded under turbulent fluid conditions in the CDC reactor on exposure to 100 mg ml(-1) of AgNPs. The antibacterial activity of AgNPs on various microbial strains grown on polycarbonate membranes is reported. In conclusion, AgNPs effectively prevent the formation of biofilms and kill bacteria in established biofilms, which suggests that AgNPs could be used for prevention and treatment of biofilm-related infections. Further research and development are necessary to translate this technology into therapeutic and preventive strategies.

Antimicrobial activity, cytotoxicity and inflammatory response of novel plastics embedded with silver nanoparticles.

AIM: Infections associated with medical devices are an important cause of morbidity and mortality. Microorganisms are responsible for catheter infections that may then result in the local or systemic dissemination of the microorganism into the bloodstream. The aim of this study was to evaluate the antimicrobial activity of silver nanoparticles (AgNPs) embedded in polyurethane plastics, commonly used for catheter fabrication. MATERIALS & METHODS: AgNPs in the range of 25-30 nm were synthesized and embedded in polyurethane plastics at different concentrations. The antimicrobial activities of these plastics were tested against the three pathogenic microorganisms, Escherichia coli, Staphylococcus epidermidis and Candida albicans, frequently associated with catheter infections. The cytotoxicity of the plastics was evaluated on human-derived macrophages using propidium iodide and the secretion of the pro- and anti-inflammatory cytokines IL-6, IL-10 and TNF-a was measured using ELISA. RESULTS: A significant reduction of 6- to 7-log in the number of bacteria was measured, while a reduction of 90% was measured in the case of C. albicans. Neither cytotoxic effect on macrophages nor immunological response was observed. CONCLUSION: Plastics embedded with AgNPs have great potential to limit microbial colonization of implanted medical devices.

Effectiveness of bonding resin-based composite to healthy and fluorotic enamel using total-etch and two self-etch adhesive systems.

The aim of this study was to evaluate the bond strength of three adhesive systems: Excite™, Adper Prompt L-Pop™ and AdheSE One™ to varying degrees of fluorotic enamel using micro-tensile bond strength (µTBS) tests. Human enamel was classified according to the Thylstrup and Fejerskov Index. The interface resin-enamel was observed using stereoscopic and electron microscopy. The Excite™, achieved the highest µTBS when bonded to healthy enamel and decreased as the degree of fluorosis increased (p<0.05). The Prompt L-Pop™ improved the bonding on moderate and severe fluorosis. The µTBS of the AdheSE One™, was significantly lower in all degrees of fluorotic enamel (p<0.05) indicating a very poor bonding ability to enamel. These results will provide clinicians with preliminary data to assist them in the selection of the most effective adhesive systems for treatment of fluorosis enamel, resulting in more successful restorative care.

Antibacterial activity, inflammatory response, coagulation and cytotoxicity effects of silver nanoparticles.

The incorporation of nanoparticles (NPs) in industrial and biomedical applications has increased significantly in recent years, yet their hazardous and toxic effects have not been studied extensively. Here, we studied the effects of 24 nm silver NPs (AgNPs) on a panel of bacteria isolated from medical devices used in a hospital intensive care unit. The cytotoxic effects were evaluated in macrophages and the expression of the inflammatory cytokines IL-6, IL-10 and TNF-α were quantified. The effects of NPs on coagulation were tested in vitro in plasma-based assays. We demonstrated that 24 nm AgNPs were effective in suppressing the growth of clinically relevant bacteria with moderate to high levels of antibiotic resistance. The NPs had a moderate inhibitory effect when coagulation was initiated through the intrinsic pathway. However, these NPs are cytotoxic to macrophages and are able to elicit an inflammatory response. Thus, beneficial and potential harmful effects of 24 nm AgNPs on biomedical devices must be weighed in further studies in vivo. From the Clinical Editor: The authors of this study demonstrate that gallic acid reduced 24 nm Ag NPs are effective in suppressing growth of clinically relevant antibiotic resistant bacteria. However, these NPs also exhibit cytotoxic properties to macrophages and may trigger an inflammatory response. Thus, the balance of beneficial and potential harmful effects must be weighed carefully in further studies.

Synergistic bactericidal activity of Ag-TiO2 nanoparticles in both light and dark conditions.

High-throughput screening was employed to evaluate bactericidal activities of hybrid Ag-TiO2 nanoparticles comprising variations in TiO2 crystalline phase, Ag content, and synthesis method. Hybrid Ag-TiO2 nanoparticles were prepared by either wet-impregnation or UV photo deposition onto both Degussa P25 and DuPont R902 TiO2 nanoparticles. The presence of Ag was confirmed by ICP, TEM, and XRD analysis. The size of Ag nanoparticles formed on anatase/rutile P25 TiO2 nanoparticles was smaller than those formed on pure rutile R902. When activated by UV light, all hybrid Ag-TiO2 nanoparticles exhibited stronger bactericidal activity than UV alone, Ag/UV, or UV/TiO2. For experiments conducted in the dark, bactericidal activity of Ag-TiO2 nanoparticles was greater than either bare TiO2 (inert) or pure Ag nanoparticles, suggesting that the hybrid materials produced a synergistic antibacterial effect unrelated to photoactivity. Moreover, less Ag(+) dissolved from Ag-TiO2 nanoparticles than from Ag nanoparticles, indicating the antibacterial activities of Ag-TiO2 was not only caused by releasing of toxic metal ions. It is clear that nanotechnology can produce more effective bactericides; however, the challenge remains to identify practical ways to take advantage of these exciting new material properties.

In vitro cytotoxicity of silver nanoparticles on human periodontal fibroblasts.

UNLABELLED: Silver nanoparticles (NNPs) are extensively used for all kinds of antimicrobial applications in medical research. Their efficacy has been demonstrated against Streptococcus mutans, which is associated with dental caries. However their cytotoxic effects on human periodontal tissue are not completely understood. OBJECTIVE: The aim of this study was to evaluate the possible toxic cellular effects of different concentrations and sizes of silver nanoparticles, less than 10 nm, 15-20 nm, and 80-100 nm, respectively, on human periodontal fibroblasts. STUDY DESIGN: Primary culture cells isolated from human periodontal tissue were exposed to 0-1,000 microM silver nanoparticles of each size for 24-, 72-, and 168-hour periods. Cytotoxicity was evaluated with a nonradioactive, soluble MTS/PMS assay. RESULTS: The results demonstrated that silver nanoparticles of less than 20 nm increased cytotoxicity in human periodontal fibroblasts in a dose- and time-dependent manner. CONCLUSION: The 80-100-nm-sized nanoparticles did not modify the viability of human primary culture cells.