Engineering of Injectable Antibiotic-laden Fibrous Microparticles Gelatin Methacryloyl Hydrogel for Endodontic Infection Ablation

This study aimed at engineering cytocompatible and injectable antibiotic-laden fibrous microparticles gelatin methacryloyl (GelMA) hydrogels for endodontic infection ablation. Clindamycin (CLIN) or metronidazole (MET) was added to a polymer solution and electrospun into fibrous mats, which were processed via cryomilling to obtain CLIN- or MET-laden fibrous microparticles. Then, GelMA was modified with CLIN- or MET-laden microparticles or by using equal amounts of each set of fibrous microparticles. Morphological characterization of electrospun fibers and cryomilled particles was performed via scanning electron microscopy (SEM). The experimental hydrogels were further examined for swelling, degradation, and toxicity to dental stem cells, as well as antimicrobial action against endodontic pathogens (agar diffusion) and biofilm inhibition, evaluated both quantitatively (CFU/mL) and qualitatively via confocal laser scanning microscopy (CLSM) and SEM. Data were analyzed using ANOVA and Tukey’s test (α = 0.05). The modification of GelMA with antibiotic-laden fibrous microparticles increased the hydrogel swelling ratio and degradation rate. Cell viability was slightly reduced, although without any significant toxicity (cell viability > 50%). All hydrogels containing antibiotic-laden fibrous microparticles displayed antibiofilm effects, with the dentin substrate showing nearly complete elimination of viable bacteria. Altogether, our findings suggest that the engineered injectable antibiotic-laden fibrous microparticles hydrogels hold clinical prospects for endodontic infection ablation.

eDNA Inactivation and Biofilm Inhibition by the Polymeric Biocide Polyhexamethylene Guanidine Hydrochloride (PHMG-Cl)

The choice of effective biocides used for routine hospital practice should consider the role of disinfectants in the maintenance and development of local resistome and how they might affect antibiotic resistance gene transfer within the hospital microbial population. Currently, there is little understanding of how different biocides contribute to eDNA release that may contribute to gene transfer and subsequent environmental retention. Here, we investigated how different biocides affect the release of eDNA from mature biofilms of two opportunistic model strains Pseudomonas aeruginosa ATCC 27853 (PA) and Staphylococcus aureus ATCC 25923 (SA) and contribute to the hospital resistome in the form of surface and water contaminants and dust particles. The effect of four groups of biocides, alcohols, hydrogen peroxide, quaternary ammonium compounds, and the polymeric biocide polyhexamethylene guanidine hydrochloride (PHMG-Cl), was evaluated using PA and SA biofilms. Most biocides, except for PHMG-Cl and 70% ethanol, caused substantial eDNA release, and PHMG-Cl was found to block biofilm development when used at concentrations of 0.5% and 0.1%. This might be associated with the formation of DNA–PHMG-Cl complexes as PHMG-Cl is predicted to bind to AT base pairs by molecular docking assays. PHMG-Cl was found to bind high-molecular DNA and plasmid DNA and continued to inactivate DNA on surfaces even after 4 weeks. PHMG-Cl also effectively inactivated biofilm-associated antibiotic resistance gene eDNA released by a pan-drug-resistant Klebsiella strain, which demonstrates the potential of a polymeric biocide as a new surface-active agent to combat the spread of antibiotic resistance in hospital settings.

Sustainable phyto-fabrication of silver nanoparticles using Gmelina arborea exhibit antimicrobial and biofilm inhibition activity

Increase in bacterial resistance to commonly used antibiotics is a major public health concern generating interest in novel antibacterial treatments. Aim of this scientific endeavor was to find an alternative efficient antibacterial agent from non-conventional plant source for human health applications. We used an eco-friendly approach for phyto-fabrication of silver nanoparticles (AgNPs) by utilizing logging residue from timber trees Gmelina arborea (GA). GC–MS analysis of leaves, barks, flowers, fruits, and roots was conducted to determine the bioactive compounds. Biosynthesis, morphological and structural characterization of GA-AgNPs were undertaken by UV–Vis spectroscopy, scanning electron microscopy (SEM), energy-dispersive spectroscopy (EDX), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR) and X-ray diffractometer (XRD). GA-AgNPs were evaluated for antibacterial, antibiofilm, antioxidant, wound healing properties and their toxicity studies were carried out. Results identified the presence of terpenoids, sterols, aliphatic alcohols, aldehydes, and flavonoids in leaves, making leaf extract the ideal choice for phyto-fabrication of silver nanoparticles. The synthesis of GA-AgNPs was confirmed by dark brown colored colloidal solution and spectral absorption peak at 420 nm. Spherical, uniformly dispersed, crystalline GA-AgNPs were 34–40 nm in diameter and stable in solutions at room temperature. Functional groups attributed to the presence of flavonoids, terpenoids, and phenols that acted as reducing and capping agents. Antibacterial potency was confirmed against pathogenic bacteria Bacillus cereus, Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus by disc diffusion assay, MIC and MBC assay, biofilm inhibition assay, electron-microscopy, cell staining and colony counting techniques. The results from zone of inhibition, number of ruptured cells and dead-cell-count analysis confirmed that GA-AgNPs were more effective than GA-extract and their bacteria inhibition activity level increased further when loaded on hydrogel as GA-AgNPs-PF127, making it a novel distinguishing feature. Antioxidant activity was confirmed by the free radical scavenging assays (DPPH and ABTS). Wound healing potential was confirmed by cell scratch assay in human dermal fibroblast cell lines. Cell-proliferation study in human chang liver cell lines and optical microscopic observations confirmed non-toxicity of GA-AgNPs at low doses. Our study concluded that biosynthesized GA-AgNPs had enhanced antibacterial, antibiofilm, antioxidant, and wound healing properties.

Antibiofilm Activities of Biogenic Silver Nanoparticles Against Candida albicans

Biofilms are microbial colonies that are encased in an organic polymeric matrix and are resistant to antimicrobial treatments. Biofilms can adhere to both biotic and abiotic surfaces, allowing them to colonize medical equipment such as urinary and intravenous catheters, mechanical heart valves, endotracheal tubes, and prosthetic joints. Candida albicans biofilm is the major etiological cause of the pathogenesis of candidiasis in which its unobstructed growth occurs in the oral cavity; trachea, and catheters that progress to systemic infections in the worst scenarios. There is an urgent need to discover novel biofilm preventive and curative agents. In the present investigation, an effort is made to observe the role of cyanobacteria-derived AgNPs as a new antibiofilm agent with special reference to candidiasis. AgNPs synthesized through the green route using Anabaena variabilis cell extract were characterized by UV–visible spectroscopy. The nanoparticles were spherical in shape with 11–15 nm size and were monodispersed. The minimum inhibitory concentration (MIC) of AgNPs was obtained at 12.5 μg/mL against C. albicans. AgNPs 25 μg/mL showed 79% fungal cell membrane permeability and 22.2% ROS production. AgNPs (25 μg/mL) also facilitated 62.5% of biofilm inhibition and degradation. Therefore, AgNPs could be considered as a promising antifungal agent to control biofilm produced by C. albicans.

The Investigation of Thymol Formulations Containing Poloxamer 407 and Hydroxypropyl Methylcellulose to Inhibit Candida Biofilm Formation and Demonstrate Improved Bio-Compatibility

The aim of this study was to investigate the potential of thymol to inhibit Candida biofilm formation and improve thymol biocompatibility in the presence of hydroxypropyl methylcellulose (HPMC) and poloxamer 407 (P407), as possible drug carriers. Thymol with and without polymers were tested for its ability to inhibit biofilm formation, its effect on the viability of biofilm and biocompatibility studies were performed on HEK 293 (human embryonic kidney) cells. Thymol showed a concentration dependent biofilm inhibition; this effect was slightly improved when it was combined with HPMC. The Thymol-P407 combination completely inhibited the formation of biofilm and the antibiofilm effect of thymol decreased as the maturation of Candida biofilms increased. The effect of thymol on HEK 293 cells was a loss of nearly 100% in their viability at a concentration of 250 mg/L. However, in the presence of P407, the viability was 25% and 85% using neutral red uptake and sulforhodamine B assays, respectively. While, HPMC had less effect on thymol activity the thymol-P407 combination showed a superior inhibitory effect on biofilm formation and better biocompatibility with human cell lines. The combination demonstrates a potential medical use for the prevention of Candida biofilm formation.

Controlling Listeria monocytogenes Growth and Biofilm Formation using Flavonoids

The aim of the present study was to investigate the ability of natural plant-derivate (flavonoid compounds) products to reduce and/or inhibit the biofilm-forming ability of Listeria monocytogenes. A collection of 500 synthetic and natural flavonoids were tested on strains of L. monocytogenes for their antimicrobial and anti-biofilm activity. L. monocytogenes biofilm inhibition by flavonoid compounds was tested on i) stainless steel coupons using crystal violet staining and ii) glass slides using confocal laser scanning microscopic (CLSM) imaging. The flavonoids were tested against a L. monocytogenes cocktail of 5 strains at a concentration of 100 µM to determine their effect on planktonic growth. A total of 17 flavonoids were chosen for further study due to their ability to significantly reduce the growth of L. monocytogenes in BHI broth, while 2 flavonoids were chosen because they actually increased growth. A lower concentration of flavonoid compounds (50 µM) was selected to investigate their effects on L. monocytogenes biofilm formation using i) stainless steel coupons to quantify biomass and ii) glass coupons to observe the biofilm architecture. The 19 flavonoids showed various levels of L. monocytogenes growth inhibition, ranging from 2% to 100%, as compared to the respective positive and negative controls on stainless steel, after 48 h of incubation at 22 o C. In addition, in comparison to the control, most of the 19 flavonoids significantly (p ≤ 0.05) inhibited biofilm formation, with at least one of the L. monocytogenes strains or at one of the tested temperatures. In fact, when grown in BHI broth with 50 µM of the 19 selected flavonoid compounds for 48 h at 22 o C, there were visible reductions in L. monocytogenes biofilm formation on the glass coupons. Overall, we found multiple flavonoid compounds to be promising anti-biofilm and antimicrobial agents against L. monocytogenes .

The Effect of the plant Adiantum philippense Extracts on Biofilms Formation and Adhesion to Shigella flexneri: A Predictive Modelling Approach

In the quest for novel bioactive metabolites, which can also be used as therapeutic agents, Adiantum philippense (A. philippense), an ethnomedically important fern, has become a fascinating herb. In this study, the predictive mathematical modelling of A. philippense crude extract was tested against Shigella flexneri, a common food pathogen for its phytochemical constituents, antagonistic ability, and effect on bacterial adhesion and biofilm formation was calculated. Various kinetics models such as Von Bertalanffy, Baranyi-Roberts, modified Schnute, Modified Richards, Modified Gompertz, Modified Logistics and latest Huang were used to get values for the above kinetic constants or parameters. modified Gompertz of the entire model was found to be the best model with the highest adjusted R2 value and lowest RMSE value. The accuracy and bias factors values were close to unity (1.0). The maximum specific growth rate (mmax (h-1) for S. flexneri treatment with A. philippense extract was significantly much lower (p<0.05) with a value of 0.292 (95% confidence interval of 0.254 to 0.331) compared to control with a value of 0.540 (95% confidence interval of 0.481 to 0.599) indicating potential biofilm inhibition.

2D PdCu Alloy Nanodendrites Manifest Effective Peroxidase-Like Activity Against Biofilms

Noble metal nanomaterials with peroxidase-like catalytic activity have received great interest lately for their potential applications in biomedicine and environmental protection; however, it is still challenging to achieve high catalytic efficiency despite enormous efforts. In this work, a novel but simple route was developed to synthesize 2D PdCu alloy nanodendrites (PdCu NDs) as a high-performance peroxidase mimic for biofilm elimination. Catalytic kinetics shows that the composition-dependent synergy between Pd and Cu in the PdCu NDs can strongly enhance the peroxidase-like activity. Density functional theory calculations further provide the underlying mechanisms at both atomic and electronic levels for the effective adsorption and dissociation of H2O2 molecules on PdCu NDs surfaces. Owing to their superior peroxidase-like activity, the PdCu NDs exhibit striking biofilm inhibition properties, which suggests that the controlled synthesis of 2D noble metal alloy may open up new opportunities for enhancing enzyme-like activities of noble metal nanomaterials.

Effect of Plasmonic Gold Nanoprisms on Biofilm Formation and Heat Shock Proteins Expression in Human Pathogenic Bacteria

Gold nanoparticles have gained interest in biomedical sciences in the areas of nano-diagnostics, bio-labeling, drug delivery, and bacterial infection. In this study, we examined, for the first time, the antibacterial and antibiofilm properties of plasmonic gold nanoprisms against human pathogenic bacteria using MIC and crystal violet. In addition, the expression level of GroEL/GroES heat shock proteins was also investigated by western blot. Gold nanoparticles were characterized by TEM and EDX, which showed equilateral triangular prisms with an average edge length of 150 nm. Antibacterial activity testing showed a great effect of AuNPs against pathogenic bacteria with MICs values ranging from 50 μg/mL to 100 μg/mL. Nanoparticles demonstrated strong biofilm inhibition action with a percentage of inhibition ranging from 40.44 to 82.43%. Western blot analysis revealed that GroEL was an AuNPs-inducible protein with an increase of up to 66.04%, but GroES was down-regulated with a reduction of up to 46.81%. Accordingly, plasmonic gold nanoprisms, could be a good candidate for antibiotics substitution in order to treat bacterial infections.