Role of nanomaterial physicochemical properties on fate and toxicity in bacteria and plants

Nanomaterials, defined as having at least one dimension <100 nm, are ubiquitous in nature. However, engineered nanomaterials have gained increasing attention for use in drug-delivery applications and consumer goods. Examination of nanomaterial toxicity, both beneficial (e.g., drug delivery to bacterial pathogens) and detrimental (e.g., death of terrestrial plants), thus warranted. Herein, I present the evaluation of nitric oxidereleasing nanomaterial toxicity to bacteria and silica particle toxicity to plants as a function of nanomaterial physicochemical properties. Nanomaterial toxicity toward planktonic (i.e., free-floating) Pseudomonas aeruginosa and Staphylococcus aureus bacteria was evaluated as a function of scaffold size, shape, and exterior functionality using nitric oxide-releasing (NO) silica particles, dendrimers, and chitosan oligosaccharides. Improved bactericidal efficacy was observed for silica particles with decreased size and increased aspect ratio due to improved particle–cell interactions. Likewise, better nanomaterial–bacteria association and biocidal action was noted for more hydrophobic NO-releasing dendrimers and chitosan oligosaccharides. Planktonic bacterial killing was not dependent on chitosan molecular weight due to rapid association between the cationic scaffolds and negatively-charged bacterial cell membranes. Given the importance of nanomaterial physicochemical properties in planktonic bacterial killing, the NO-releasing scaffolds were also evaluated against clinicallyrelevant bacterial biofilms. Similar to planktonic studies, smaller particle sizes proved more efficient in delivering NO throughout the biofilm. Particles with rod-like shape also eradicated biofilms more effectively. The role of NO-releasing dendrimer and chitosan oligosaccharide hydrophobicity was prominent in scaffold diffusion through the biofilm and subsequent NO delivery, with scaffolds modified with hydrophobic functionalities generally exhibiting better bacterial association. Lastly, biofilm eradication was more effective for NO-releasing dendrimers exhibiting sustained NO-release compared to delivery of NO via an initial burst. Phytotoxicity and uptake of silica nanoparticles was evaluated for the plant Arabidopsis thaliana as a function of particle size, surface composition, and shape (i.e., spherical versus rod-like particles). Overall, the silica nanoparticles examined were found to be relatively non-toxic to A. thaliana plants when pH effects were mitigated. Sizedependent uptake of the silica particles was observed, with smaller particles concentrating more heavily in the roots, rosette, and stem; however no shape-dependent uptake was noted at the low exposure concentration examined.Doctor of Philosoph

Nitric oxide-releasing chitosan oligosaccharides as antibacterial agents

Secondary amine-functionalized chitosan oligosaccharides of different molecular weights (i.e., ~2500, 5000, 10000) were synthesized by grafting 2-methyl aziridine from the primary amines on chitosan oligosaccharides, followed by reaction with nitric oxide (NO) gas under basic conditions to yield N-diazeniumdiolate NO donors. The total NO storage, maximum NO flux, and half-life of the resulting NO-releasing chitosan oligosaccharides were controlled by the molar ratio of 2-methyl aziridine to primary amines (e.g., 1:1, 2:1) and the functional group surrounding the N-diazeniumdiolates (e.g., polyethylene glycol (PEG) chains), respectively. The secondary amine-modified chitosan oligosaccharides greatly increased the NO payload over existing biodegradable macromolecular NO donors. In addition, the water-solubility of the chitosan oligosaccharides enabled their penetration across the extracellular polysaccharides matrix of Pseudomonas aeruginosa biofilms and association with embedded bacteria. The effectiveness of these chitosan oligosaccharides at biofilm eradication was shown to depend on both the molecular weight and ionic characteristics. Low molecular weight and cationic chitosan oligosaccharides exhibited rapid association with bacteria throughout the entire biofilm, leading to enhanced biofilm killing. At concentrations resulting in 5-log killing of bacteria in Pseudomonas aeruginosa biofilms, the NO-releasing and control chitosan oligosaccharides elicited no significant cytotoxicity to mouse fibroblast L929 cells in vitro

Nitric Oxide-Releasing Amphiphilic Poly(amidoamine) (PAMAM) Dendrimers as Antibacterial Agents

A series of amphiphilic nitric oxide (NO)-releasing poly(amidoamine) (PAMAM) dendrimers with different exterior functionalities were synthesized by a ring-opening reaction between primary amines on the dendrimer and propylene oxide (PO), 1,2-epoxy-9-decene (ED), or a ratio of the two, followed by reaction with NO at 10 atm to produce N-diazeniumdiolate-modified scaffolds with a total storage of ~1 μmol/mg. The hydrophobicity of the exterior functionality was tuned by varying the ratio of PO and ED grafted onto the dendrimers. The bactericidal efficacy of these NO-releasing vehicles against established Gram-negative Pseudomonas aeruginosa biofilms was then evaluated as a function of dendrimer exterior hydrophobicity (i.e., ratio of PO/ED), size (i.e., generation), and NO release. Both the size and exterior functionalization of dendrimer proved important to a number of parameters including dendrimer-bacteria association, NO delivery efficiency, bacteria membrane disruption, migration within the biofilm, and toxicity to mammalian cells. Although enhanced bactericidal efficacy was observed for the hydrophobic chains (e.g., ED), toxicity to L929 mouse fibroblast cells was also noted at concentrations necessary to reduce bacterial viability by 5-logs (99.999% killing). The optimal PO to ED ratios for biofilm eradication with minimal toxicity against L929 mouse fibroblast cells were 7:3 and 5:5. The study presented herein demonstrated the importance of both dendrimer size and exterior properties in determining efficacy against established biofilms without compromising biocompatibility to mammalian cells

Nitric Oxide-Releasing Silica Nanoparticle-Doped Polyurethane Electrospun Fibers

Electrospun polyurethane fibers doped with nitric oxide (NO)-releasing silica particles are presented as novel macromolecular scaffolds with prolonged NO-release and high porosity. Fiber diameter (119–614 nm) and mechanical strength (1.7–34.5 MPa of modulus) were varied by altering polyurethane type and concentration, as well as the NO-releasing particle composition, size, and concentration. The resulting NO-releasing electrospun nanofibers exhibited ~83% porosity with flexible plastic or elastomeric behavior. The use of N-diazeniumdiolate- or S-nitrosothiol-modified particles yielded scaffolds exhibiting a wide range of NO release totals and durations (7.5 nmol mg−1–0.12 μmol mg−1 and 7 h to 2 weeks, respectively). The application of NO-releasing porous materials as coating for subcutaneous implants may improve tissue biocompatibility by mitigating the foreign body response and promoting cell integration

Influence of Scaffold Size on Bactericidal Activity of Nitric Oxide-Releasing Silica Nanoparticles

A reverse microemulsion synthesis was used to prepare amine functionalized silica nanoparticles of three distinct sizes (i.e., 50, 100, and 200 nm) with identical amine concentrations. The resulting hybrid nanoparticles, consisting of N-(6 aminohexyl) aminopropyltrimethoxysilane and tetraethoxysilane, were highly monodisperse in size. N-diazeniumdiolate nitric oxide (NO) donors were subsequently formed on secondary amines while controlling reaction conditions to keep the total amount of nitric oxide (NO) released constant for each particle size. The bactericidal efficacy of the NO releasing nanoparticles against Pseudomonas aeruginosa increased with decreasing particle size. Additionally, smaller diameter nanoparticles were found to associate with the bacteria at a faster rate and to a greater extent than larger particles. Neither control (non-NO-releasing) nor NO releasing particles exhibited toxicity towards L929 mouse fibroblasts at concentrations above their respective minimum bactericidal concentrations. This study represents the first investigation of the bactericidal efficacy of NO-releasing silica nanoparticles as a function of particle size

Dual Action Antimicrobials: Nitric Oxide Release from Quaternary Ammonium-Functionalized Silica Nanoparticles

The synthesis of quaternary ammonium (QA)-functionalized silica nanoparticles with and without nitric oxide (NO) release capabilities is described. Glycidyltrialkylammonium chlorides of varied alkyl chain lengths (i.e., methyl, butyl, octyl, and dodecyl) were tethered to the surface of amine-containing silica nanoparticles via a ring-opening reaction. Secondary amines throughout the particle were then functionalized with N-diazeniumdiolates NO donors to yield dual functional nanomaterials with surface QAs and total NO payloads of ca. 0.3 μmol/mg. The bactericidal activities of singly (i.e., only NO-releasing or only QA-functionalized) and dual (i.e., NO-releasing and QA-functionalized) functional nanoparticles were tested against Grampositive Staphylococcus aureus and Gram-negative Pseudomonas aeruginosa. Particles with only NO release capabilities alone were found to be more effective against P. aeruginosa, while particles with only QA-functionalities exhibited greater toxicity toward S. aureus. The minimum bactericidal concentrations (MBC) of QA-functionalized particles decreased with increasing alkyl chain length against both microbes tested. Combining NO release and QA-functionalities on the same particle resulted in an increase in bactericidal efficacy against S. aureus; however, no change in activity against P. aeruginosa was observed compared to NO-releasing particles alone

Nitric Oxide-Releasing Dendrimers as Antibacterial Agents

The antibacterial activity of a series of nitric oxide (NO)-releasing poly(propylene imine) (PPI) dendrimers was evaluated against both Gram-positive and Gram-negative pathogenic bacteria, including methicillin-resistant Staphylococcus aureus. A direct comparison of the bactericidal efficacy between NO-releasing and control PPI dendrimers (i.e., non-NO-releasing) revealed both enhanced biocidal action of NO-releasing dendrimers and reduced toxicity against mammalian fibroblast cells. Antibacterial activity for the NO donor-functionalized PPI dendrimers was shown to be a function of both dendrimer size (molecular weight) and exterior functionality. In addition to minimal toxicity against fibroblasts, NO-releasing PPI dendrimers modified with styrene oxide exhibited the greatest biocidal activity (≥9.999% killing) against all bacterial strains tested. The N-diazeniumdiolate NO donor-functionalized PPI dendrimers presented in this study hold promise as effective NO-based therapeutics for combating bacterial infections

Fate of TiO2 nanoparticles in the aquatic environment in the presence of anthropogenic compounds

International audienceThe increasing production and use of nanoparticles (NP) in consumer products inevitably lead to ENP emissions into the environment. The physicochemical properties of NP depend on various parameters (e.g. pH, cations, IS). In natural waters, the stability of NP can vary as a function of a sum of these parameters and occurs by one of the numerous scenarios. In particular, the presence of anthropogenic organic molecules (AOM) can change the NP fate. Also, the presence of NP may affect the organic pollutants (fate and toxicity). The main objective of the work was to study the aggregation of TiO2 NP (pure hydrophilic 100 % rutile and pure hydrophilic 100 % anatase, 5−30 nm) in the presence of the most frequently occur and representative pesticides (glyphosate, AMPA, 2.4D) in natural waters considering lab experiments under relevant aqueous conditions (pH, ionic strength, presence and concentrations of mono- and bivalent cations). The presence of pesticides affected TiO2 NP homoaggregation in solutions (IS=10-3M - 10-2M) with pH values below the NP point of zero charge (PZC) for the anatase NPs (pH=6.5) and with pH values above the NP PZC for the rutile NP (pH=4.5). No changes in NP aggregation were observed in very low (IS=10-4M) or very high (IS= 10-1M) ionic strength solutions. The presence of the pesticides caused a significant modification of the NP surface charge (zeta potential) over a large range of salt concentrations (IS=10-4M - 10-1M). Compared to mono-valent cations (Na+), bi-valent cations (Ca2+) favor an increase in zeta potential of NP (anatase and rutile) at pH 8. There is no significant difference between at pH 5. Finally, these results demonstrated that, among the studied AOMs, glyphosate (with 4 pKa-s from 0.8 to 11) affects NP aggregation/stabilization in a wider range of physicochemical conditions. Overall, these results will aid in the evaluation of potential environmental risks posed by engineered NP in the aquatic environments exposed to pesticide load

The effect of nitric oxide surface flux on the foreign body response to subcutaneous implants

Although the release of nitric oxide (NO) from biomaterials has been shown to reduce the foreign body response (FBR), the optimal NO release kinetics and doses remain unknown. Herein, polyurethane-coated wire substrates with varying NO release properties were implanted into porcine subcutaneous tissue for 3, 7, 21 and 42 d. Histological analysis revealed that materials with short NO release durations (i.e., 24 h) were insufficient to reduce the collagen capsule thickness at 3 and 6 weeks, whereas implants with longer release durations (i.e., 3 and 14 d) and greater NO payloads significantly reduced the collagen encapsulation at both 3 and 6 weeks. The acute inflammatory response was mitigated most notably by systems with the longest duration and greatest dose of NO release, supporting the notion that these properties are most critical in circumventing the FBR for subcutaneous biomedical applications (e.g., glucose sensors)

Release and fate of nanoparticulate TiO2 UV filters from sunscreen: Effects of particle coating and formulation type

International audienceNanoparticulate mineral UV filters, such as titanium dioxide (TiO 2) nanocomposites, are being increasingly used in sunscreens as an alternative to organic UV filters. However, there is still a lack of understanding regarding their fate and behavior in aquatic environments and potential environmental impacts after being released from a bather's skin during recreational activities. In this work, we assessed the release, fate, and transformation of two commercial nanocomposite TiO 2 UV filters, one hydrophobic and one hydrophilic, in ultrapure water and simulated fresh-and seawater. The hydrophobic TiO 2 nanocomposite, T-SA, was coated with a primary Al 2 O 3 photopassivation layer and a secondary stearic acid layer, while the hydrophilic TiO 2 nanocomposite, T-SiO 2 , was coated with a single SiO 2 photopassivation layer. The influence of the sunscreen formulation was examined by dispersing the TiO 2 nanocomposites in their typical continuous phase (i.e., oil for T-SA and water for T-SiO 2) before introduction into the aqueous system. After 48 h of aqueous aging and 48 h of settling, 88e99% of the hydrophobic T-SA remained floating on top of the water column in all aqueous systems. On the other hand, 100% of the hydrophilic T-SiO 2 settled out of the water column in the fresh-and seawaters. With respect to the photopassivation coatings, no loss of the T-SA Al 2 O 3 layer was detected after aqueous aging, but 99e100% dissolution of the SiO 2 layer on the T-SiO 2 nanocomposite was observed after 48 h in the fresh-and seawaters. This dissolution left behind T-SiO 2 by-products exhibiting a photocatalytic activity similar to that of bare rutile TiO 2. Overall, the results demonstrated that the TiO 2 surface coating and sunscreen formulation type drive environmental behavior and fate and that loss of the passivation layer can result in potentially harmful, photoactive by-products. These insights will help guide regulations and assist manufacturers in developing more environmentally safe sunscreens