In vitro and in vivo evaluation of cerium oxide nanoparticles in respiratory syncytial virus infection.

Respiratory syncytial virus (RSV) is the most common cause of viral bronchiolitis among children worldwide, yet there is no vaccine for RSV disease. This study investigates the potential of cube and sphere-shaped cerium oxide nanoparticles (CNP) to modulate reactive oxygen (ROS) and nitrogen (RNS) species and immune cell phenotypes in the presence of RSV infection in vitro and in vivo. Cube and sphere-shaped CNP were synthesized by hydrothermal and ultrasonication methods, respectively. Physico-chemical characterization confirmed the shape of sphere and cube CNP and effect of various parameters on their particle size distribution and zeta potential. In vitro results revealed that sphere and cube CNP differentially modulated ROS and RNS levels in J774 macrophages. Specifically, cube CNP significantly reduced RSV-induced ROS levels without affecting RNS levels while sphere CNP increased RSV-induced RNS levels with minimal effect on ROS levels. Cube CNP drove an M1 phenotype in RSV-infected macrophages in vitro by increasing macrophage surface expression of CD80 and CD86 with a concomitant increase in TNFα and IL-12p70, while simultaneously decreasing M2 CD206 expression. Intranasal administration of sphere and cube-CNP were well-tolerated with no observed toxicity in BALB/c mice. Notably, cube CNP preferentially accumulated in murine alveolar macrophages and induced their activation, avoiding enhanced uptake and activation of other inflammatory cells such as neutrophils, which are associated with RSV-mediated inflammation. In conclusion, we report that sphere and cube CNP modulate macrophage polarization and innate cellular responses during RSV infection.

Altering the interfacial rheology of Pseudomonas aeruginosa and Staphylococcus aureus with N-acetyl cysteine and cysteamine.

Introduction: Chronic lung infection due to bacterial biofilms is one of the leading causes of mortality in cystic fibrosis (CF) patients. Among many species colonizing the lung airways, Pseudomonas aeruginosa and Staphylococcus aureus are two virulent pathogens involved in mechanically robust biofilms that are difficult to eradicate using airway clearance techniques like lung lavage. To remove such biological materials, glycoside hydrolase-based compounds are commonly employed for targeting and breaking down the biofilm matrix, and subsequently increasing cell susceptibility to antibiotics. Materials and methods: In this study, we evaluate the effects of N-acetyl cysteine (NAC) and Cysteamine (CYST) in disrupting interfacial bacterial films, targeting different components of the extracellular polymeric substances (EPS). We characterize the mechanics and structural integrity of the interfacial bacterial films using pendant drop elastometry and scanning electron microscopy. Results and discussion: Our results show that the film architectures are compromised by treatment with disrupting agents for 6 h, which reduces film elasticity significantly. These effects are profound in the wild type and mucoid P. aeruginosa, compared to S. aureus. We further assess the effects of competition and cooperation between S. aureus and P. aeruginosa on the mechanics of composite interfacial films. Films of S. aureus and wild-type P. aeruginosa cocultures lose mechanical strength while those of S. aureus and mucoid P. aeruginosa exhibit improved storage modulus. Treatment with NAC and CYST reduces the elastic property of both composite films, owing to the drugs' ability to disintegrate their EPS matrix. Overall, our results provide new insights into methods for assessing the efficacy of mucolytic agents against interfacial biofilms relevant to cystic fibrosis infection.

Mucoid Coating Provides a Growth Advantage to Pseudomonas aeruginosa at Oil-Water Interfaces.

Chronic lung infection with bacterial biofilms is one of the leading causes of death in cystic fibrosis (CF) patients. Among many species infecting the lung airways, Pseudomonas aeruginosa is the major pathogen colonizing and persisting throughout the patient's life. The microorganism undergoes pathoadaptation, while switching from a nonmucoid to a mucoid phenotype, improving the mechanical properties of the resulting biofilms. Previous investigation of the dynamic rheological properties of nonmucoid (PANT) and mucoid (PASL) clinical P. aeruginosa isolates exposed to interfacial stresses demonstrated that the mucoid strains formed films with stronger resistance to bending and nonlinear relaxation to compression and tension. We hypothesize that the mucoid switch provides a growth advantage to P. aeruginosa through the development of interfacial films with viscoelastic properties enabling cell survival. Here, we investigate the physiological response of the mucoid and the nonmucoid P. aeruginosa to interfacial entrapment. Our results, both macroscopic and molecular, reveal that mucoid coating plays an important role in protecting the bacteria from interfacial stresses. Cell characterizations using electron and fluorescence microscopies showed higher proportion of dead nonmucoid cells compared to mucoid cells on interfacial exposure. For example, scanning transmission electron microscopy (STEM) imaging showed that 96.6% of nonmucoid cells vs only 22.2% of mucoid cells were lysed owing to interfacial stress. Furthermore, the transcriptional profiling of P. aeruginosa cells indicated the upregulation of pel, psl, and alginate genes encoding for exopolysaccharide biomaterials is associated with mucoid cells' ability to cope with the interfacial environments. Further characterization of real-time gene regulation at interfaces will elucidate the effects of interfacial environment on the regulation of bacterial virulence.

Material properties of interfacial films of mucoid and nonmucoid Pseudomonas aeruginosa isolates.

Chronic lung infection with bacterial biofilms is a leading cause of death in cystic fibrosis (CF) patients. Pseudomonas aeruginosa, one of the many species colonizing the lung airways, can undergo pathoadaptation, leading to a mucoid phenotype with interesting material properties. We hypothesize that the surface properties and extracellular materials of mucoid P. aeruginosa cells greatly influence the mechanical behavior of their films at fluid interfaces. In this study, we investigate the interfacial properties of films formed by nonmucoid (PANT) and mucoid (PASL) strains of P. aeruginosa isolated from CF patients. We use pendant drop elastometry to analyze the interfacial response of the films formed by PANT and PASL at the hexadecane-water interface. The dynamic rheological analyses of the films highlight the distinctive signature of the mucoid strains at fluid interfaces. The mucoid films exhibit greater relaxation following a compressive strain than a tensile one, while a full hysteresis response is achieved by the nonmucoid films; this indicates that the material properties of the PANT films are conserved under both compression and tension. The wrinkling and shape analyses of the interfacial bacterial films elucidate that the mucoid strain exhibits remarkable viscoelastic properties, enabling the remodeling of the living films and dissipation of the compressive stress. The comparative analysis of the material properties of mucoid and nonmucoid P. aeruginosa cells indicates that mucoid switch can play an important role in protecting the bacteria from interfacial stresses. Further characterization of interfacial bacterial films will provide new insights into the development of methods for controlling interfacial films of bacteria.

Electrochemical Strategy for Eradicating Fluconazole-Tolerant Candida albicans Using Implantable Titanium.

A persistent problem in modern health care derives from the overwhelming presence of antibiotic-resistant microbes on biomaterials, more specifically, fungal growth on metal-based implants. This study seeks to investigate the antifungal properties of low-level electrochemical treatments delivered using titanium electrodes against Candida albicans. We show that C. albicans can be readily controlled with electrical currents/potentials, reducing the number of viable planktonic cells by 99.7% and biofilm cells by 96.0-99.99%. Additionally, this study explores the ability of the electrochemical treatments to potentiate fluconazole, a clinically used antifungal drug. We have found that electrochemical treatment substantially enhances fluconazole killing activity. While fluconazole alone exhibits a low efficiency against the stationary phase and biofilm cells of C. albicans, complete eradication corresponding to 7-log killing is achieved when the antifungal drug is provided subsequently to the electrochemical treatment. Further mechanistic analyses have revealed that the sequential treatment shows a complex multimodal action, including the disruption of cell wall integrity and permeability, impaired metabolic functions, and enhanced susceptibility to fluconazole, while altering the biofilm structure. Altogether, we have developed and optimized a new therapeutic strategy to sensitize and facilitate the eradication of fluconazole-tolerant microbes from implantable materials. This work is expected to help advance the use of electrochemical approaches in the treatment of infections caused by C. albicans in both nosocomial and clinical cases.

Synthesis, physico-chemical characterization, and antioxidant effect of PEGylated cerium oxide nanoparticles.

Cerium oxide nanoparticles (CNPs) represent a promising class of antioxidant nanoparticles with potential therapeutic value. Due to the easily reversible oxidation states of cerium (Ce3+ and Ce4+) at the nanoscale, CNPs scavenge excessive reactive oxygen and nitrogen species in a self-regenerative manner. In this study, we have demonstrated a simple method to functionalize shape-specific CNPs (i.e., rod- and cube-shaped) with polyethylene glycol (PEG) and studied the effect of PEGylation on the physico-chemical properties, antioxidant activity, and biocompatibility of rod- and cube-shaped CNPs. The chemical conjugation of PEG onto the CNP surface was confirmed by a series of physico-chemical characterizations (1H-NMR, FTIR, and surface zeta potential). Rod-shaped CNPs demonstrated greater reactive oxygen species scavenging ability compared to cube-shaped CNPs. PEGylation of CNPs did not affect shape, cerium oxidation state, and cytocompatibility. Importantly, PEGylation significantly reduced the amount of proteins adsorbed onto the CNPs. The antioxidant effects of CNPs were maintained in PEGylated CNPs. We envision that PEGylated rod-shaped CNPs synthesized in this study have the potential to be biocompatible nanoparticles that can combat oxidative stress-related diseases.

Effect of surfactant in mitigating cadmium oxide nanoparticle toxicity: Implications for mitigating cadmium toxicity in environment.

Cadmium (Cd), classified as human carcinogen, is an extremely toxic heavy metal pollutant, and there is an increasing environmental concern for cadmium exposure through anthropogenic sources including cigarette smoke. Though Cd based nanoparticles such as cadmium oxide (CdO) are being widely used in a variety of clinical and industrial applications, the toxicity of CdO nanoparticles has not been well characterized. Herein we report the toxicity of CdO nanoparticles employing zebrafish as a model. Two different CdO nanoparticles were prepared, calcination of Cd(OH)2 without any organic molecule (CdO-1) and calcination of Cd-citrate coordination polymer (CdO-2), to evaluate and compare the toxicity of these two different CdO nanoparticles. Results show that zebrafish exposed to CdO-2 nanoparticles expressed reduced toxicity as judged by lower oxidative stress levels, rescue of liver carboxylesterases and reduction in metallothionein activity compared to CdO-1 nanoparticles. Histopathological observations also support our contention that CdO-1 nanoparticles showed higher toxicity relative to CdO-2 nanoparticles. The organic unit of Cd-citrate coordination polymer might have converted into carbon during calcination that might have covered the surface of CdO nanoparticles. This carbon surface coverage can control the release of Cd2+ ions in CdO-2 compared to non-covered CdO-1 nanoparticles and hence mitigate the toxicity in the case of CdO-2. This was supported by atomic absorption spectrophotometer analyses of Cd2+ ions release from CdO-1 and CdO-2 nanoparticles. Thus the present study clearly demonstrates the toxicity of CdO nanoparticles in an aquatic animal and also indicates that the toxicity could be substantially reduced by carbon coverage. This could have important implications in terms of anthropogenic release and environmental pollution caused by Cd and human exposure to Cd2+ from sources such as cigarette smoke.