Jiun-Chiou Wei

Evaluation on Cytotoxicity and Genotoxicity of the Exfoliated Silicate Nanoclay

The concern about toxicity for nanosilicate platelets (NSP) derived from natural montmorillonite clay is addressed. The NSP nanoclay was isolated from polyamine-salt exfoliation of the layered silicate clay into randomized individual plates, possessing multiple ionic charges on the surface of silicate plates with an average geometric dimension of ca. 80 x 80 x 1 nm(3). The material had been previously shown to be effective for antimicrobial and tendency for adhering onto the biomaterial surface based on the direct observation by using scanning electron microscope. The material safety on genotoxic effect was investigated by using three different test systems: the Comet assay test on Chinese Hamster Ovary (CHO) cells in vitro, micronucleus (MN) assay in vivo and the Salmonella gene mutation assay on strain TA98, TA100, TA102, TA1535 and TA1537. The Comet assay showed no DNA damage after 24 h of incubation with NSP of 1000 microg/mL. The MN test indicated no significant micronucleus induction in the CHO cells at the concentrations tested. With all five strains of Salmonella typhimurium, none of mutations was found. Furthermore, cytotoxicity of the same material was assayed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) release, showing a low cytotoxicity on CHO cells below 1000 microg/mL after 12 h incubation period and a dose-dependent effect after 24 h incubation. For feeding to rats, the acute oral toxicity was shown a low lethal dose (LD(50)) or greater than 5700 mg/kg body weight for both male and female Sprague-Dawley rats. Overall, the study has demonstrated the safety of the NSP for potential uses in biomedical areas.

Novel nanohybrids of silver particles on clay platelets for inhibiting silver-resistant bacteria

We develop a novel nanohybrid showing a strong antibacterial activity on all of the tested pathogens, including methicillin-resistant Staphylococcus auerus and silver-resistant E. coli. The nanohybrid consists of silver nanoparticles (AgNPs) supported on 1 nm-thick silicate platelets (NSPs). The AgNP/NSP nanohybrid enables to encapsulate bacteria and triggers death signals from the cell membrane. The geographic shape of the NSPs concentrates AgNPs but impedes their penetration into attached cells, mitigating the detrimental effect of silver ion deposition in applied tissues. Moreover, the tightly tethered AgNPs on NSP surface achieve a stronger biocidal effect than silver nitrate, but bypassing Ag+ mechanism, on silver-resistant bacteria. This nanohybrid presents an effective and safe antimicrobial agent in a new perspective.

Efficacy and safety of nanohybrids comprising silver nanoparticles and silicate clay for controlling Salmonella infection

Developing effective and safe drugs is imperative for replacing antibiotics and controlling multidrug-resistant microbes. Nanoscale silicate platelet (NSP) and its nanohybrid, silver nanoparticle/NSP (AgNP/NSP), have been developed, and the nanohybrids show a strong and general antibacterial activity in vitro. Here, their efficacy for protecting Salmonella-infected chicks from fatality and septicemia was evaluated. Both orally administrated NSP and AgNP/NSP, but not AgNPs alone, effectively reduced the systemic Salmonella infection and mortality. In addition, quantitative Ag analyses demonstrated that Ag deposition from AgNP/NSP in the intestines was less than that from conventional AgNPs, indicating that the presence of NSP for immobilizing AgNPs reduced Ag accumulation in tissue and improved the safety of AgNPs. These in vivo results illustrated that both NSP and AgNP/NSP nanohybrid represent potential agents for controlling enteric bacterial infections.

Nanohybrids of Silver Particles Immobilized on Silicate Platelet for Infected Wound Healing

Silver nanoparticles supported on nanoscale silicate platelets (AgNP/NSP) possess interesting properties, including a large surface area and high biocide effectiveness. The nanohybrid of AgNP/NSP at a weight ratio 7/93 contains 5-nm Ag particles supported on the surface of platelets with dimensions of approximately 80×80×1 nm3. The nanohybrid expresses a trend of lower cytotoxicity at the concentration of 8.75 ppm Ag and low genotoxicity. Compared with conventional silver ions and the organically dispersed AgNPs, the nanohybrid promotes wound healing. We investigated overall wound healing by using acute burn and excision wound healing models. Tests on both infected wound models of mice were compared among the AgNP/NSP, polymer-dispersed AgNPs, the commercially available Aquacel, and silver sulfadiazine. The AgNP/NSP nanohybrid was superior for wound appearance, but had similar wound healing rates, vascular endothelial growth factor (VEGF)-A levels and transforming growth factor (TGF)-β1 expressions to Aquacel and silver sulfadiazine.

Label-free and culture-free microbe detection by three dimensional hot-junctions of flexible Raman-enhancing nanohybrid platelets

Novel nanohybrid arrays of silver (Ag)-on-silicate platelets with flexibility and three-dimensional (3D) hot-junctions (particularly in z-direction) were discovered for improving the stability of free nanoparticles and the mobility of rigid (glass or silicon-based) substrates in surface-enhanced Raman scattering (SERS) detection technology. Since the Ag nanoparticles are adsorbed on both sides of few nanometer-thick silicate platelets (single-layer exfoliated clay), the geometric arrangement of Ag on both sides of the nanoplatelets (Ag/NSP) may induce strong hot-junctions (z-direction) in reference to the pristine montmorillonite clay (multi-layers) at the thickness of ∼20 nm, measured by small molecules (adenine of DNA) and bacteria (S. aureus). Enormous red-shifts (16 nm wavelength difference) were observed between single layer and multi-layer silicate platelets, showing that huge surface plasmon enhancement comes from hot junctions in the z-direction (∼7 times higher than 2D hot-junctions of traditional SERS biochips). Further, the Ag/NSP SERS substrate displays a free floating mobility and optical transparency (less background interference), which inherently increase the contacted surface-area between the substrate and microorganisms, to enhance the SERS sensitivity. The surface modulation with a surfactant could be complimentary towards a variety of microorganisms including hydrophobic microbes, irregular-shaped microorganisms and larger biological cells due to their mutual specific surface interactions. It was anticipated to apply in the rapid detection for varied microbes with label-free and culture-free characterizations.

Selective SERS Detecting of Hydrophobic Microorganisms by Tricomponent Nanohybrids of Silver–Silicate-Platelet–Surfactant

Nanohybrids consisting of silver nanoparticles (Ag), clay platelets, and a nonionic surfactant were prepared and used as the substrate for surface-enhanced Raman scattering (SERS). The nanoscale silicate platelets (SP) (with dimensions of 100 × 100 nm(2) and a thickness of ∼1 nm) were previously prepared from exfoliation of the natural layered silicates. The tricomponent nanohybrids, Ag-SP-surfactant (Ag-SP-S), were prepared by in situ reduction of AgNO3 in the presence of clay and the surfactant. The clay platelets with a large surface area and ionic charge (ca. 18 000 sodium ions per platelet) allowed for the stabilization of Ag nanoparticles in the range of 10-30 nm in diameter. With the addition of a nonionic surfactant such as poly(oxyethylene) alkyl ether, the tricomponent Ag-SP-S nanohybrids possessed an altered affinity for contacting microorganisms. The particle size and interparticle gaps between neighboring Ag on SP were characterized by TEM. The surface tension of Ag-SP and Ag-SP-S in water implied different interactions between Ag and hydrophobic bacteria ( Escherichia coli and Mycobacterium smegmatis ). By increasing the surfactant content in Ag-SP-S, the SERS peak intensity was dramatically enhanced compared to the Ag-SP counterpart. The nanohybrids, Ag-SP and Ag-SP-S, with the advantages of varying hydrophobic affinity, floating in medium, and 3D hot-junction enhancement could be tailored for use as SERS substrates. The selective detection of hydrophobic microorganisms and larger biological cells makes SERS a possible rapid, label-free, and culture-free method of biodetection.

Enhancing silver nanoparticle and antimicrobial efficacy by the exfoliated clay nanoplatelets

The antimicrobial efficacy of silver nanoparticles (AgNP) is enhanced by using the exfoliated clay nanoscale silicate platelets (NSP) as the supports. The NSP were in the dimension of 80 nm × 80 nm × 1 nm and were derived from the exfoliation of natural clays such as sodium montmorillonite. Increasing the amount of NSP addition at the relative Ag/silicate composition ratios from 0.5/99.5 to 50/50 w/w significantly altered the particle size AgNP from 3.6 nm to 35 nm in diameter. Evaluation of antibacterial activities for these nanohybrids demonstrated the effect of the amount of NSP support and Ag size on inhibiting the growth of dermal pathogens including Gram-positive and Gram-negative bacteria. Scanning electron microscope observations further revealed the interaction of the AgNP/NSP nanohybrids in contact with individual bacteria surface. The tailoring of the AgNP/NSP nanostructures has largely affected the bacterial surface interaction by NSP adherence and hence the antimicrobial efficacy.

Surfactant-Modified Nanoclay Exhibits an Antiviral Activity with High Potency and Broad Spectrum

ABSTRACT Nanomaterials have the characteristics associated with high surface-to-volume ratios and have been explored for their antiviral activity. Despite some success, cytotoxicity has been an issue in nanomaterial-based antiviral strategies. We previously developed a novel method to fully exfoliate montmorillonite clay to generate the most fundamental units of nanoscale silicate platelet (NSP). We further modified NSP by capping with various surfactants and found that the surfactant-modified NSP (NSQ) was less cytotoxic. In this study, we tested the antiviral potentials of a series of natural-clay-derived nanomaterials. Among the derivatives, NSP modified with anionic sodium dodecyl sulfate (NSQc), but not the pristine clay, unmodified NSP, a silver nanoparticle-NSP hybrid, NSP modified with cationic n-octadecanylamine hydrochloride salt, or NSP modified with nonionic Triton X-100, significantly suppressed the plaque-forming ability of Japanese encephalitis virus (JEV) at noncytotoxic concentrations. NSQc also blocked infection with dengue virus (DEN) and influenza A virus. Regarding the antiviral mechanism, NSQc interfered with viral binding through electrostatic interaction, since its antiviral activity can be neutralized by Polybrene, a cationic polymer. Furthermore, NSQc reduced the lethality of JEV and DEN infection in mouse challenge models. Thus, the surfactant-modified exfoliated nanoclay NSQc may be a novel nanomaterial with broad and potent antiviral activity. IMPORTANCE Nanomaterials have being investigated as antimicrobial agents, yet their antiviral potential is overshadowed by their cytotoxicity. By using a novel method, we fully exfoliated montmorillonite clay to generate the most fundamental units of nanoscale silicate platelet (NSP). Here, we show that the surfactant-modified NSP (NSQ) is less cytotoxic and that NSQc (NSP modified with sodium dodecyl sulfate) could potently block infection by dengue virus (DEN), Japanese encephalitis virus (JEV), and influenza A virus at noncytotoxic concentrations. For the antiviral mechanism, we find that the electrostatic interaction between the negatively charged NSQc and the positively charged virus particles blocks viral binding. Furthermore, we used mouse challenge models of JEV and DEN to demonstrate the in vivo antiviral potential of NSQc. Thus, NSQc may function as a potent and safe antiviral nanohybrid against several viruses, and our success in synthesizing surfactant-modified NSP with antiviral activity may shed some light on future antiviral development.