Polyethylene terephthalate nanoparticles effect on RAW 264.7 macrophage cells

AbstractPlastic pollution is a major environmental concern due to its pervasiveness which continues to increase year on year, as a result of a continuing acceleration in global plastic production and use. Polyethylene terephthalate (PET) is among the most produced plastics, commonly used as food and beverage containers. Once released in the environment, the degradation of plastic materials produces micro-and nano-plastics, with a particular concern about potential toxicological effects if they cross epithelial barriers via inhalation or ingestion. In this work, the effect of PET nanoparticles (PET-NPs) (≤ 250 d.nm) was assayed on mouse macrophages cell line (RAW 264.7) in in vitro experiments. Results showed that PET nanoparticles were easily internalized by the cells, 15 μg/mL of nanoparticles concentration had exhibited effects in cell proliferation and a slightly increased production of Reactive Oxygen Species (ROS), which seems to trigger cell response as foreign particles related to upregulation of PCDH12, IGH-V10, ROBO1 genes, and cell maintenance functions, related to FTSJ2 gene upregulation. Thus, the RAW 264.7 results showed here are useful towards for a preliminary and understanding of the potentially toxic effects related to PET nanoparticles and complementary to other in vitro assays, as the first step into the development of the risk assessment framework

Effect of antimicrobial nanocomposites on Vibrio cholerae lifestyles: Pellicle biofilm, planktonic and surface-attached biofilm.

Vibrio cholerae is an important human pathogen causing intestinal disease with a high incidence in developing countries. V. cholerae can switch between planktonic and biofilm lifestyles. Biofilm formation is determinant for transmission, virulence and antibiotic resistance. Due to the enhanced antibiotic resistance observed by bacterial pathogens, antimicrobial nanomaterials have been used to combat infections by stopping bacterial growth and preventing biofilm formation. In this study, the effect of the nanocomposites zeolite-embedded silver (Ag), copper (Cu), or zinc (Zn) nanoparticles (NPs) was evaluated in V. cholerae planktonic cells, and in two biofilm states: pellicle biofilm (PB), formed between air-liquid interphase, and surface-attached biofilm (SB), formed at solid-liquid interfaces. Each nanocomposite type had a distinctive antimicrobial effect altering each V. cholerae lifestyles differently. The ZEO-AgNPs nanocomposite inhibited PB formation at 4 μg/ml, and prevented SB formation and eliminated planktonic cells at 8 μg/ml. In contrast, the nanocomposites ZEO-CuNPs and ZEO-ZnNPs affect V. cholerae viability but did not completely avoid bacterial growth. At transcriptional level, depending on the nanoparticles and biofilm type, nanocomposites modified the relative expression of the vpsL, rbmA and bap1, genes involved in biofilm formation. Furthermore, the relative abundance of the outer membrane proteins OmpT, OmpU, OmpA and OmpW also differs among treatments in PB and SB. This work provides a basis for further study of the nanomaterials effect at structural, genetic and proteomic levels to understand the response mechanisms of V. cholerae against metallic nanoparticles

Synthesis and Complete Antimicrobial Characterization of CEOBACTER, an Ag-Based Nanocomposite.

The antimicrobial activity of silver nanoparticles (AgNPs) is currently used as an alternative disinfectant with diverse applications, ranging from decontamination of aquatic environments to disinfection of medical devices and instrumentation. However, incorporation of AgNPs to the environment causes collateral damage that should be avoided. In this work, a novel Ag-based nanocomposite (CEOBACTER) was successfully synthetized. It showed excellent antimicrobial properties without the spread of AgNPs into the environment. The complete CEOBACTER antimicrobial characterization protocol is presented herein. It is straightforward and reproducible and could be considered for the systematic characterization of antimicrobial nanomaterials. CEOBACTER showed minimal bactericidal concentration of 3 μg/ml, bactericidal action time of 2 hours and re-use capacity of at least five times against E. coli cultures. The bactericidal mechanism is the release of Ag ions. CEOBACTER displays potent bactericidal properties, long lifetime, high stability and re-use capacity, and it does not dissolve in the solution. These characteristics point to its potential use as a bactericidal agent for decontamination of aqueous environments

Enhancement of antibiotics antimicrobial activity due to the silver nanoparticles impact on the cell membrane.

The ability of microorganisms to generate resistance outcompetes with the generation of new and efficient antibiotics; therefore, it is critical to develop novel antibiotic agents and treatments to control bacterial infections. An alternative to this worldwide problem is the use of nanomaterials with antimicrobial properties. Silver nanoparticles (AgNPs) have been extensively studied due to their antimicrobial effect in different organisms. In this work, the synergistic antimicrobial effect of AgNPs and conventional antibiotics was assessed in Gram-positive and Gram-negative bacteria. AgNPs minimal inhibitory concentration was 10-12 μg mL-1 in all bacterial strains tested, regardless of their different susceptibility against antibiotics. Interestingly, a synergistic antimicrobial effect was observed when combining AgNPs and kanamycin according to the fractional inhibitory concentration index, FICI: <0.5), an additive effect by combining AgNPs and chloramphenicol (FICI: 0.5 to 1), whereas no effect was found with AgNPs and β-lactam antibiotics combinations. Flow cytometry and TEM analysis showed that sublethal concentrations of AgNPs (6-7 μg mL-1) altered the bacterial membrane potential and caused ultrastructural damage, increasing the cell membrane permeability. No chemical interactions between AgNPs and antibiotics were detected. We propose an experimental supported mechanism of action by which combinatorial effect of antimicrobials drives synergy depending on their specific target, facilitated by membrane alterations generated by AgNPs. Our results provide a deeper understanding about the synergistic mechanism of AgNPs and antibiotics, aiming to combat antimicrobial infections efficiently, especially those by multi-drug resistant microorganisms, in order to mitigate the current crisis due to antibiotic resistance

Bactericidal parameters (expressed as μg/ml) of the CEOBACTER nanocomposite compared with those of Zeolite-Ag nanocomposites and colloidal AgNPs.

<p>Bactericidal parameters (expressed as μg/ml) of the CEOBACTER nanocomposite compared with those of Zeolite-Ag nanocomposites and colloidal AgNPs.</p

Histogram for the Ag nanoparticles size (A) and high-resolution Ag 3d (B) and O 1s (C) XPS spectra of the CEOBACTER.

<p>Solid line curve correspond to full multi-Gaussian fit, and dash-dot line curves show the deconvoluted signals.</p

AgNPs-ion release is sufficient to exert a bactericidal effect over <i>E</i>. <i>coli</i> cultures.

<p>By dialysis experiments, confined bacteria was killed by ions released from CEOBACTER (A). No release of AgNPs were shown in culture media inside and outside dialysis sacks (B). Ag ions was measured by ICP analysis. Free colloidal AgNPs at reported concentrations were used as a sensitivity control.</p

Exposure time and agent concentration dependence of bactericidal effect of CEOBACTER.

<p>The values are the average of three different experiments.</p

XRD analysis.

<p>XRD patterns of the CEOBACTER, the Na-mordenite (NaMOR) and the standard NaMOR ICSD 68445 file [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0166205#pone.0166205.ref026" target="_blank">26</a>]. Inside, the NaMOR unit cell is shown.</p

Bacterial survival values (in percentage) for different CEOBACTER concentrations at increasing number of inoculums.

<p>Bacterial survival values (in percentage) for different CEOBACTER concentrations at increasing number of inoculums.</p