Effectiveness of plant-based hand sanitizer incorporating Quercus infectoria gall extract.

AIMS: Quercus infectoria (Qi), a traditional herbal plant with a broad spectrum of activities on multidrug-resistant bacteria, has been developed for hand sanitizer applications. METHODS AND RESULTS: Antimicrobial activity was evaluated using agar-well diffusion and broth microdilution method. Bactericidal activity was determined following the European Standard 1276 antibacterial suspension test. Neutralization assay was performed to assess antirespiratory syncytial virus. Safety, stability, and skin permeation of Qi hand gel was investigated. Qi hand sanitizer gel inhibited microorganisms ranging from 99.9% to 99.999% against Enterococcus faecalis, Staphylococcus aureus, methicillin-resistant Staph. aureus, Staph. epidermidis, Staph. pseudintermedius, Staph. saprophyticus, Streptococcus pyogenes, Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Candida albicans. A significant reduction in main human dermatophytes including Microsporum canis, M. gypseum, and Talaromyces marneffei of ∼50% was observed (P < .05). Qi hand sanitizer gel inactivated >99% viral particles entering human laryngeal epidermoid carcinoma cells in a dose-dependent manner. Scanning electron micrographs further illustrated that Qi hand sanitizer gel disrupted microbial cell membrane after 1-min contact time resulting in cell death. Qi hand sanitizer gel delivered emollient compounds through simulated human skin layers and showed no cytotoxicity on fibroblast cells. Moreover, Qi hand sanitizer gel demonstrated stability under extreme conditions. CONCLUSIONS: Qi hand sanitizer gel was able to inhibit various microorganisms including bacteria, dermatophytes, and virus.

Biogenic nanosilver-fabricated endotracheal tube to prevent microbial colonization in a veterinary hospital.

COVID-19 patients have often required prolonged endotracheal intubation, increasing the risk of developing ventilator-associated pneumonia (VAP). A preventive strategy is proposed based on an endotracheal tube (ETT) modified by the in situ deposition of eucalyptus-mediated synthesized silver nanoparticles (AgNPs). The surfaces of the modified ETT were embedded with AgNPs of approximately 28 nm and presented a nanoscale roughness. Energy dispersive X-ray spectroscopy confirmed the presence of silver on and inside the coated ETT, which exhibited excellent antimicrobial activity against Gram-positive and Gram-negative bacteria, and fungi, including multidrug-resistant clinical isolates. Inhibition of planktonic growth and microbial adhesion ranged from 99 to 99.999% without cytotoxic effects on mammalian cells. Kinetic studies showed that microbial adhesion to the coated surface was inhibited within 2 h. Cell viability in biofilms supplemented with human tracheal mucus was reduced by up to 95%. In a porcine VAP model, the AgNPs-coated ETT prevented adhesion of Pseudomonas aeruginosa and completely inhibited bacterial invasion of lung tissue. The potential antimicrobial efficacy and safety of the coated ETT were established in a randomized control trial involving 47 veterinary patients. The microbial burden was significantly lower on the surface of the AgNPs-coated ETT than on the uncoated ETT (p < 0.05). KEY POINTS: • Endotracheal tube surfaces were modified by coating with green-synthesized AgNPs • P. aeruginosa burden of endotracheal tube and lung was reduced in a porcine model • Effective antimicrobial activity and safety was demonstrated in a clinical trial.

Biologically rapid synthesized silver nanoparticles from aqueous Eucalyptus camaldulensis leaf extract: Effects on hyphal growth, hydrolytic enzymes, and biofilm formation in Candida albicans.

Bionanotechnology has increasingly gained attention in biomedical fields as antifungal and antibiofilm agents. In this study, biosynthesized silver nanoparticles (bio-AgNPs) using aqueous Eucalyptus camaldulensis leaf extract were successfully performed by a one-step green approach. Spherical-shaped nanoparticles, approximately 8.65 nm, exhibited noncytotoxicity to erythrocytes, HeLa, and HaCaT cells. The synthesized nanoparticles showed strong fungicidal activity ranging from 0.5 to 1 µg/ml. The nanoparticles affected Candida adhesion and invasion into host cells by reduced germ tube formation and hydrolytic enzyme secretion. Inhibitory effects of bio-AgNPs on Candida biofilms were evaluated by the prevention of yeast-to-hyphal transition. A decrease in cell viability within mature biofilm demonstrated the ability of bio-AgNPs to penetrate into the extracellular matrix and destroy yeast cell morphology, leading to cell death. Molecular biology study on biofilms confirmed downregulation in the expression of genes ALS3, HWP1, ECE1, EFG1, TEC1, ZAP1, encoding hyphal growth and biofilm development and PLB2, LIP9, SAP4, involved in hydrolytic enzymes. In addition to candida treatment, the bio-AgNPs could be applied as an antioxidant to protect against oxidative stress-related human diseases. The findings concluded that bio-AgNPs could be used as an antifungal agent for candida treatment, as well as be incorporated in medical devices to prevent biofilm formation.

Prolonged inhibitory effects against planktonic growth, adherence, and biofilm formation of pathogens causing ventilator-associated pneumonia using a novel polyamide/silver nanoparticle composite-coated endotracheal tube.

Microbial cells can rapidly form biofilm on endotracheal tubes (ETT) causing ventilator-associated pneumonia, a serious complication in patients receiving mechanical ventilation. A novel polyamide with a good balance of hydrophilic/hydrophobic moieties was used for the embedment of green-reduction silver nanoparticles (AgNPs) for the composite-coated ETT. The films were conformal with a thickness of ∼ 17 ± 3 µm accommodating high loading of 60 ± 35 nm spherical-shaped AgNPs. The coated ETT resulted in a significant difference in reducing both planktonic growth and microbial adhesion of single and mixed-species cultures, compared with uncoated ETT (p < 0.05). A time-kill assay demonstrated rapid bactericidal effects of the coating on bacterial growth and cell adhesion to ETT surface. Biofilm formation by Pseudomonas aeruginosa and Staphylococcus aureus, commonly encountered pathogens, was inhibited by > 96% after incubation for 72 h. Polyamide/AgNP composite-coated ETT provided a broad-spectrum activity against both Gram-positive and Gram-negative bacteria as well as Candida albicans and prolonged antimicrobial activity.

In vitro and in vivo comparative analysis of antibacterial activity of green-synthesized silver nanoparticles.

This study aims to compare antibacterial effects of green-synthesized silver nanoparticles (AgNPs) with silver nitrate (AgNO3 ). AgNPs were successfully synthesized using Eucalyptus camaldulensis leaf extract as a reducing and stabilizing agent. Minimum inhibitory concentrations (MIC) of AgNPs and AgNO3 against Staphylococcus aureus and Pseudomonas aeruginosa ranged between 4.8 and 6.75 µg mL-1 . Growth curves demonstrated that inhibition of P. aeruginosa occurred right after AgNPs were added and throughout the period of the study (72 h). Antibacterial effects of both AgNPs and AgNO3 could be abrogated by cysteine and 2-mercaptoethanol, thiol-containing compounds. Galleria mellonella model revealed relatively low toxic effects of both AgNPs and AgNO3 . At 20MIC of AgNPs (≈137.8 mg kg-1 ), more than 80% survival of G. mellonella was observed. Unexpectedly, silver-containing agents could not rescue larvae after S. aureus infection. Further ex vivo experiments in the presence of coelomic larval fluid demonstrated the reduction of antibacterial activity of both AgNPs and AgNO3 . It was speculated that anionic molecules present in the coelomic fluid might neutralize the action of Ag ions. Binding of AgNPs or AgNO3 to albumin, a major protein in human blood which transport several endogenous compounds was not detected, indicating that the silver-containing agents could be applied as an antimicrobial agent.

Control of biomaterial-associated infections through biofabrication of gold nanoparticles using Musa sapientum extract.

Biofilm-associated infections are a critical element in infectious diseases and play an important role in antibiotic resistance. Biosynthesized gold nanoparticles (AuNPs) using ethanolic extract of Musa sapientum unripe fruit were performed. The nanoparticles demonstrated an absorption peak at 554 nm with particle sizes ranging from 5.45 to 104.44 nm. High negative zeta potential value of -33.97 mV confirmed the high stability of AuNPs. The presence of bioconstituents responsible for capping and stabilization was indicated by intensity changes of several peaks from Fourier-transform infrared spectroscopy analysis. The minimum inhibitory concentrations (MIC) of the biosynthesized AuNPs against important pathogens ranged from 10 to 40 µg mL-1 . Synthesized nanoparticles at 0.062 to 0.5 × MIC significantly inhibited biofilm formation in all the tested microorganisms (p < 0.05). Scanning electron microscopy and confocal scanning laser microscopy images clearly illustrated in disruption and architectural changes of microbial biofilms at sub-MIC of biosynthesized AuNPs. Excellent antioxidant and antityrosinase activities of AuNPs were observed. The biosynthesized AuNPs at 20 µg mL-1 significantly inhibited nitric oxide production by 93% in lipopolysaccharide-stimulated RAW 264.7 cells, compared with control (p < 0.05). The biosynthesized AuNPs at 0.6 to 40 µg mL-1 demonstrated no toxic effects on L929 fibroblast cells.

Eucalyptus-Mediated Synthesized Silver Nanoparticles-Coated Urinary Catheter Inhibits Microbial Migration and Biofilm Formation.

Catheter-associated urinary tract infections (CAUTIs) are significant complications among catheterized patients, resulting in increased morbidity, mortality rates, and healthcare costs. Foley urinary catheters coated with synthesized silver nanoparticles (AgNPs) using Eucalyptus camaldulensis leaf extract were developed using a green chemistry principle. In situ-deposited AgNPs with particle size ranging between 20 and 120 nm on the catheter surface were illustrated by scanning electron microscopy. Atomic force microscopy revealed the changes in surface roughness after coating with nanoparticles. The coated catheter could significantly inhibit microbial adhesion and biofilm formation performed in pooled human urine-supplemented media to mimic a microenvironment during infections (p 0.05). AgNPs-coated catheter exhibited broad-spectrum antimicrobial activity against important pathogens, causing CAUTIs with no cytotoxic effects on HeLa cells. A reduction in microbial viability in biofilms was observed under confocal laser scanning microscopy. A catheter bridge model demonstrated complete prevention of Proteus mirabilis migration by the coated catheter. Significant inhibition of ascending motility of Escherichia coli and P. mirabilis along the AgNPs-coated catheter was demonstrated in an in vitro bladder model (p 0.05). The results suggested that the AgNPs-coated urinary catheter could be applied as an alternative strategy to minimize the risk of CAUTIs by preventing bacterial colonization and biofilm formation.

Poly (vinyl alcohol) copolymerized with xanthan gum/hypromellose/sodium carboxymethyl cellulose dermal dressings functionalized with biogenic nanostructured materials for antibacterial and wound healing application.

Effective treatment of infected wounds requires a comprehensive wound dressing with a combination of antibacterial, antioxidative, and anti-inflammatory effects. Biodegradable wound dressings incorporating nanostructured material were developed using polyvinyl alcohol with xanthan gum, hypromellose, or sodium carboxymethyl cellulose and extensively evaluated for antibacterial and wound healing efficacy. Synthesized silver nanoparticles and wound dressings displayed λmax at 420 nm with zeta potential ≈ - 35 mV. Significant growth inhibition with >99 % reduction in CFU/ml (p < 0.05) against important wound pathogens including Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus epidermidis, and Candida albicans were observed. Within 1 h of treatment, hypromellose nanocomposite demonstrated excellent bactericidal effects with a 99.9 % of reduction in growth. In addition, wound dressings demonstrated inhibitory activities against free radical scavengers. Wound dressings demonstrated a significant reduction in the inflammatory response in RAW 264.7 macrophages (p < 0.001). Ex-vivo diffusion demonstrated zero-order release and steady-state flux between 0.1571-0.2295 µg/ml/cm2h with 0.124-0.144 permeability coefficient after 10 h. Usage in animals further confirmed that the hypromellose nanocomposite accelerated the wound healing process with biocompatibility. The results suggested that hybrid biodegradable dressings can be effectively applied to treat infected wounds and attenuate inflammatory responses.

Prevention of nosocomial transmission and biofilm formation on novel biocompatible antimicrobial gloves impregnated with biosynthesized silver nanoparticles synthesized using Eucalyptus citriodora leaf extract.

Failure in the prevention of cross-transmission from contaminated gloves has been recognized as an important factor that contributes to the spread of several healthcare-associated infections. Ex situ coating process with silver nanoparticles (AgNPs) using Eucalyptus citriodora ethanolic leaf extract as reducing and capping agents to coat glove surfaces has been developed to prevent this mode of transmission. Elemental analysis of coated gloves showed 24.8 Wt% silver densely adhere on the surface. The coated gloves fully eradicated important hospital-acquired pathogens including Gram-positive bacteria, Gram-negative bacteria, and yeasts within 1 h. The coated gloves showed significant reduction, an average of five logs when tested against all standard strains and most clinical isolates (p < 0.01). Following prolonged exposure, the coating significantly reduced the numbers of most adhered pathogenic species, compared with uncoated gloves (p < 0.0001). AgNPs-coated gloves reduced microbial adhesion of mixed-species biofilms. A series of contamination and transmission assays demonstrated no transmission of viable organisms. Biocompatibility analysis confirmed high viability of HaCaT and L929 cells at all concentrations of AgNPs tested. The coated gloves were non-toxic with direct contact with L929 cells. The highly efficacious AgNPs-coated gloves potentially provide additional protection against transmission of healthcare-associated infections.

Green fabrication of anti-bacterial biofilm layer on endotracheal tubing using silver nanoparticles embedded in polyelectrolyte multilayered film.

Endotracheal tubes (ETTs) are a common source of bacterial colonization, leading to ventilator-associated pneumonia (VAP). This research developed a biofilm-resistant ETT, following the principles of green chemistry. Using an aqueous layer-by-layer (LbL) technique, a thick polyelectrolyte multilayered film was deposited on a ventilation tube. The polyelectrolyte multilayered film accommodated silver nanoparticles (AgNPs) formed in situ by reducing Ag+ ions with Eucalyptus citriodora leaf extract. The multilayered film coating conformed to the curved surfaces of the ETT. Film thickness and silver content increased exponentially with the number of polyelectrolyte bilayer pairs, and a sufficiently high AgNPs content of 10-30%w/w was obtained at 75 to 125 bilayer films. Adhesion of the Gram-positive Staphylococcus aureus and Gram-negative Pseudomonas aeruginosa was prevented by 99.9 and 99.99%, respectively, without cytotoxic effects against human lung epithelial cells (p < 0.05).

Evaluation of phytochemicals from medicinal plants of Myrtaceae family on virulence factor production by Pseudomonas aeruginosa.

Virulence factors regulated by quorum sensing (QS) play a critical role in the pathogenesis of an opportunistic human pathogen, Pseudomonas aeruginosa in causing infections to the host. Hence, in the present work, the anti-virulence potential of the medicinal plant extracts and their derived phytochemicals from Myrtaceae family was evaluated against P. aeruginosa. In the preliminary screening of the tested medicinal plant extracts, Syzygium jambos and Syzygium antisepticum demonstrated a maximum inhibition in QS-dependent violacein pigment production by Chromobacterium violaceum DMST 21761. These extracts demonstrated an inhibitory activity over a virulence factor, pyoverdin, production by P. aeruginosa ATCC 27853. Gas chromatography-mass spectrometric (GC-MS) analysis revealed the presence of 23 and 12 phytochemicals from the extracts of S. jambos and S. antisepticum respectively. Three top-ranking phytochemicals, including phytol, ethyl linoleate and methyl linolenate, selected on the basis of docking score in molecular docking studies lowered virulence factors such as pyoverdin production, protease and haemolytic activities of P. aeruginosa to a significant level. In addition, the phytochemicals reduced rhamnolipid production by the organism. The work demonstrated an importance of plant-derived compounds as anti-virulence drugs to conquer P. aeruginosa virulence towards the host.