Antibiofilm effect of biogenic silver nanoparticle alone and combined with polymyxin B against carbapenem-resistant Acinetobacter baumannii.

Acinetobacter baumannii is a bacteria associated with nosocomial infections and outbreaks, difficult to control due to its antibiotic resistance, ability to survive in adverse conditions, and biofilm formation adhering to biotic and abiotic surfaces. Therefore, this study aimed to evaluate the antibiofilm activity of biogenic silver nanoparticle (Bio-AgNP) and polymyxin B alone and combined in biofilms formed by isolates of carbapenem-resistant A. baumannii (CR-Ab). In the biofilm formation inhibition assay, CR-Ab strains were exposed to different concentrations of the treatments before inducing biofilm formation, to determine the ability to inhibit/prevent bacterial biofilm formation. While in the biofilm rupture assay, the bacterial biofilm formation step was previously carried out and the adhered cells were exposed to different concentrations of the treatments to evaluate their ability to destroy the bacterial biofilm formed. All CR-Ab isolates and ATCC® 19606™ used in this study are strong biofilm formers. The antibiofilm activity of Bio-AgNP and polymyxin B against CR-Ab and ATCC® 19606™ demonstrated inhibitory and biofilm-disrupting activity. When used in combination, Bio-AgNP and polymyxin B inhibited 4.9-100% of biofilm formation in the CR-Ab isolates and ATCC® 19606™. Meanwhile, when Bio-AgNP and polymyxin B were combined, disruption of 6.8-77.8% of biofilm formed was observed. Thus, antibiofilm activity against CR-Ab was demonstrated when Bio-AgNP was used alone or in combination with polymyxin B, emerging as an alternative in the control of CR-Ab strains.

The synergic and addictive activity of biogenic silver nanoparticle associated with meropenem against carbapenem-resistant Acinetobacter baumannii.

AIMS: Antibiotic management of infections caused by Acinetobacter baumannii often fails due to antibiotic resistance (especially to carbapenems) and biofilm-forming strains. Thus, the objective here was to evaluate in vitro the antibacterial and antibiofilm activity of biogenic silver nanoparticle (Bio-AgNP) combined with meropenem, against multidrug-resistant isolates of A. baumannii. METHODS AND RESULTS: In this study, A. baumannii ATCC® 19606™ and four carbapenem-resistant A. baumannii (Ab) strains were used. The antibacterial activity of Bio-AgNP and meropenem was evaluated through broth microdilution. The effect of the Bio-AgNP association with meropenem was determined by the checkboard method. Also, the time-kill assay and the integrity of the bacterial cell membrane were evaluated. Furthermore, the antibiofilm activity of Bio-AgNP and meropenem alone and in combination was determined. Bio-AgNP has antibacterial activity with minimum inhibitory concentration (MIC) and minimum bactericidal concentration ranging from 0.46 to 1.87 µg ml-1. The combination of Bio-AgNP and meropenem showed a synergistic and additive effect against Ab strains, and Bio-AgNP was able to reduce the MIC of meropenem from 4- to 8-fold. Considering the time-kill of the cell, meropenem and Bio-AgNP when used in combination reduced bacterial load to undetectable levels within 10 min to 24 h after treatment. Protein leakage was observed in all treatments evaluated. When combined, meropenem/Bio-AgNP presents biofilm inhibition for Ab2 isolate and ATCC® 19606™, with 21% and 19%, and disrupts the biofilm from 22% to 50%, respectively. The increase in nonviable cells in the biofilm can be observed after treatment with Bio-AgNP and meropenem in carbapenem-resistant A. baumannii strains. CONCLUSIONS: The combination of Bio-AgNP with meropenem can be a therapeutic option in the treatment of infections caused by carbapenem-resistant A. baumannii.

Biogenic silver nanoparticles: in vitro activity against Staphylococcus aureus methicillin-resistant (MRSA) and multidrug-resistant coagulase-negative Staphylococcus (CoNS).

Multidrug-resistant (MDR) bacteria are one problem in health since the therapeutic alternative are reduced. For this, the application of nanotechnology through functionalized nanoparticles, like a biogenic silver nanoparticle (Bio-AgNP), obtained by biological synthesis, emerges as a possible alternative against the MDR bacteria. This study aimed to evaluate the antibacterial and antibiofilm activity of Bio-AgNP obtained for biological synthesis by Fusarium oxysporum strain 551 against methicillin-resistant Staphylococcus aureus (MRSA) and MDR coagulase-negative Staphylococcus (CoNS) isolates. Bio-AgNP has activity against S. aureus ATCC 25904, Staphylococcus epidermidis ATCC 35984, and MDR isolates, with minimal inhibitory concentration (MIC) ranging from 3.75 to 15 µg.mL-1 and minimal bactericidal concentration (MBC) from 7.5 to 30 µg.mL-1. In the membrane leakage assay, it was observed that all concentrations tested led to proteins release from the cellular content dose-dependently, where the highest concentrations led to higher protein in the supernatant. The 2×MIC of Bio-AgNP killed ATCC 35984 after 6h of treatment, and ATCC 25904 and S. aureus (SA3) strains after 24h of treatment. The 4×MIC was bactericidal in 6h of treatment for all strains in the study. The biofilm of MDR isolates was inhibited in 80.94 to 100% and eradicated in 60 to 94%. The confocal laser scanning microscopy (CLSM) analysis demonstrated similar results to the antibiofilm assays. The Bio-AgNP has antibacterial and antibiofilm activity and can be a promising therapeutic alternative against MDR bacteria.

Antibacterial and antibiofilm activity of 1-thiocarbamoyl-3,5-diaryl-4,5-dihydro-1H pyrazoles and thiazoles in multidrug-resistant pathogens.

To find novel antibiotic drugs, six 1-thiocarbamoyl-3,5-diaryl-4,5-dihydro-1H derivatives named 1b, 1d (pyrazoles), 2a, 2b, 2c, and 2d (thiazoles) were evaluated in silico and in vitro. The in silico analyses were based on ADME pharmacokinetic parameters (absorption, distribution, metabolism, and excretion). The in vitro antibacterial activity was evaluated in Gram-positive and Gram-negative species (Staphylococcus aureus ATCC® 25904, Staphylococcus epidermidis ATCC® 35984, Klebsiella pneumoniae ATCC® 700603, and Acinetobacter baumannii ATCC® 19606), by determination of minimal inhibitory concentration (MIC), minimal bactericidal concentration (MBC), kinetics curve, and antibiofilm assays. As results, the azoles have activity against the Gram-negative species K. pneumoniae ATCC® 700603 and A. baumannii ATCC® 19606. No antibacterial activity was observed for the Gram-positive bacteria evaluated. Thus, the azoles were evaluated against clinical isolates of K. pneumoniae carbapenemase (KPC) and A. baumannii multidrug-resistant (Ab-MDR). All azoles have antibacterial activity against Ab-MDR isolates (Gram-negative) with MIC values between 512 µg/mL and 1,024 µg/mL. Against KPC isolates the azoles 1b, 1d, and 2d present antibacterial activity (MIC = 1,024 µg/mL). In the kinetics curve assay, the 1b and 1d pyrazoles reduced significantly viable cells of Ab-MDR isolates and additionally inhibited 86.6 to 95.8% of the biofilm formation. The in silico results indicate high possibility to permeate the blood-brain barrier (2b) and was predict human gastrointestinal absorption (all evaluated azoles). Considering that the research and development of new antibiotics is a priority for drug-resistant pathogens, our study revealed the antibacterial and antibiofilm activity of novel azoles against K. pneumoniae and A. baumannii pathogens.

Silver nanoparticles (AgNPs) in the control of Staphylococcus spp.

The Staphylococcus bacteria cause several infections, S. aureus is the major species, expressing different virulence factors. Therefore, coagulase-negative Staphylococcus (CoNS) are nosocomial pathogens, mainly associated with biofilm formation in invasive medical devices. Methicillin-resistant S. aureus (MRSA) and multidrug resistant (MDR) CoNS are widely distributed in the hospital environment, leading to infections that are difficult to treat. Thus, nanoparticles (NPs) are studied as an alternative in the control of these pathogens. Silver nanoparticles (AgNPs) stand out due to their different biological properties, broad-spectrum antibacterial activity, low toxicity, and use in combination with other drugs. Several studies with AgNPs evaluated in-vitro against S. aureus and MRSA validated the spectrum of action of the NPs. However, few studies attempted to explore the response of the CoNS, mainly in vivo studies. Research that explored the in vivo application of AgNPs against these bacteria helped to understand and better elucidate their activity on the skin through different biological models. Furthermore, the application of NPs is a viable alternative for controlling these bacteria, including MDR bacteria, in cases of skin infections by avoiding worsening the clinical condition and favoring tissue regeneration of the injured area.

Biogenic silver nanoparticle (Bio-AgNP) has an antibacterial effect against carbapenem-resistant Acinetobacter baumannii with synergism and additivity when combined with polymyxin B.

AIMS: Carbapenem-resistant Acinetobacter baumannii represents a public health problem, and the search for new antibacterial drugs has become a priority. Here, we investigate the antibacterial activity of biogenic silver nanoparticles (Bio-AgNPs) synthesized by Fusarium oxysporum, used alone or in combination with polymyxin B against carbapenem-resistant A. baumannii. METHODS AND RESULTS: In this study, ATCC® 19606™ strain and four carbapenem-resistant A. baumannii strains were used. The antibacterial activity of Bio-AgNPs and its synergism with polymyxin B were determined using broth microdilution, checkboard methods and time-kill assays. The integrity of the bacterial cell membrane was monitored by protein leakage assay. In addition, the cytotoxicity in the VERO mammalian cell line was also evaluated, and the selectivity index was calculated. Bio-AgNPs have an antibacterial activity with MIC and MBC ranging from 0.460 to 1.870 µg/ml. The combination of polymyxin B and Bio-AgNPs presents synergy against four of the five strains tested and additivity against one strain in the checkerboard assay. Considering the time of cell death, Bio-AgNPs killed all carbapenem-resistant isolates and ATCC® 19606™ within 1 h. When combined, Bio-AgNPs presented 16-fold reduction of the polymyxin B MIC and showed a decrease in terms of viable A. baumannii cells in 4 h of treatment, with synergic and additive effects. Protein leakage was observed with increasing concentrations for Bio-AgNPs treatments. Additionally, Bio-AgNP and polymyxin B showed dose-dependent cytotoxicity against mammalian VERO cells and combined the cytotoxicity which was significantly reduced and presented a greater pharmacological safety. CONCLUSIONS: The results presented here indicate that Bio-AgNPs in combination with polymyxin B could represent a good alternative in the treatment of carbapenem-resistant A. baumannii. SIGNIFICANCE AND IMPACT OF STUDY: This study demonstrates the synergic effect between Bio-AgNPs and polymyxin B on carbapenem-resistant A. baumannii strains.

Effect of Calcium Hydroxide on Bonding Performance of an Experimental Self-etch Adhesive.

PURPOSE: To investigate the effect of Ca(OH)2 concentration on pH neutralization, degree of conversion (DC%), and bonding performance of experimental self-etch adhesives (SEAs). MATERIALS AND METHODS: Four different concentrations of Ca(OH)2 (0 wt%, 1 wt%, 2 wt%, and 4 wt%) were added to the bond of an experimental two-step SEA consisting of primer (10-MDP [30 wt%], TEG-DMA [30 wt%], ethanol [35 wt%], water [5 wt%], camphorquinone [0.5 wt%], and tertiary amine [0.5 wt%]) and bond (bis-GMA [50 wt%], TEG-DMA [30 wt%], HEMA [20 wt%], camphorquinone [0.5 wt%], and tertiary amine [0.5 wt%]) to form four groups: E0, E1, E2 and E4. pH neutralization was evaluated until it reached equilibrium, and DC% within the hybrid layer was analyzed by micro-Raman spectroscopy. Human molars were wet ground until the occlusal dentin was exposed, SEAs were applied, and composite buildups were constructed. After storage in distilled water at 37°C for 24 h, the teeth were cut into composite-dentin beams. Microtensile bond strength (µTBS) was evaluated after 24 h of water storage at 37°C. Nanoleakage was evaluated by SEM. Data were analyzed using ANOVA and Tukey's HSD test (a = 0.05). RESULTS: All the SEAs reached pH equilibrium after thirteen days, with E1 and E4 presenting the highest pH (p < 0.05). E0 and E1 presented lower DC% than did E2 and E4 (p < 0.05). All the SEAs showed statistically similar mTBS and nanoleakage (p > 0.05). CONCLUSION: The incorporation of Ca(OH)2 endowed the SEAs with pH-neutralization ability and improved their DC%, without interfering with the bond strength to dentin or nanoleakage extent.

Effect of HEMA Phosphate as an Alternative to Phosphoric Acid for Dentin Treatment Prior to Hybridization with Etch-and-Rinse Adhesive Systems.

PURPOSE: To evaluate the effect of dentin treatment using HEMA phosphate (HEMA-P) on the microtensile bond strength (µTBS) and nanoleakage of an etch-and-rinse adhesive system. MATERIALS AND METHODS: The occlusal surfaces of human molars were wet ground until superficial dentin was exposed. The specimens were then assigned to two groups according to dentin treatment: PA: 37% H3PO4 for 15 s; or HP: HEMA-P for 15 s. Adper Single Bond 2 was applied to the treated dentin surfaces and resin composite buildups were incrementally constructed over them. After 24-h storage in artificial saliva at 37°C, the bonded teeth were cut into resin-dentin sticks with a cross-sectional area of 1 mm², which were submitted to µTBS testing immediately or after 3 months of storage in artificial saliva at 37°C. Nanoleakage was assessed using SEM/EDS, and the interaction between dentin and H3PO4 or HEMA-P was evaluated by combining micro-Raman and FT-IR spectroscopy. The data were analyzed using two-way ANOVA and Tukey's HSD post-hoc test (α = 0.05). RESULTS: HP presented significantly higher µTBS than PA at both times (p < 0.05). Both treatments maintained µTBS stability after 3 months of artificial saliva storage (p > 0.005). At both times, PA presented higher nanoleakage than HP (p < 0.05). CONCLUSIONS: Both dentin treatments maintained µTBS stability after 3 months of artificial saliva storage. The use of HEMA-P was associated with less nanoleakage than was traditional phosphoric-acid etching.