Long-Term Antibacterial Efficacy of Cetylpyridinium Chloride-Montmorillonite Containing PMMA Resin Cement.

Despite being able to adhesively restore teeth, adhesives and cement do not possess any anticariogenic protection potential, by which caries recurrence may still occur and reduce the clinical lifetime of adhesive restorations. Several antibacterial agents have been incorporated into dental adhesives and cement to render them anticariogenic. Due to an additional therapeutic effect, such materials are classified as 'dental combination products' with more strict market regulations. We incorporated cetylpyridinium chloride (CPC), often used for oral hygiene applications, into montmorillonite (CPC-Mont), the latter to serve as a carrier for controlled CPC release. CPC-Mont incorporated into tissue conditioner has been approved by the Pharmaceuticals and Medical Devices Agency (PmontMDA) in Japan. To produce a clinically effective dental cement with the antibacterial potential to prevent secondary caries, we incorporated CPC-Mont into PMMA resin cement. We measured the flexural strength, shear bond strength onto dentin, CPC release, and the biofilm-inhibition potential of the experimental CPC-Mont-containing PMMA cement. An 8 and 10 wt% CPC-Mont concentration revealed the antibacterial potential without reducing the mechanical properties of the PMMA cement.

Controlled release, antimicrobial activity, and oral mucosa irritation of cetylpyridinium chloride-montmorillonite incorporated in a tissue conditioner.

This study examined the controlled release of cetylpyridinium chloride (CPC) from a tissue conditioner (TC) containing CPC-montmorillonite (CPC-Mont), the associated antimicrobial activity, and oral mucosa irritation. The CPC release test was performed daily for 28 days in three test solutions: distilled water, 0.2 M NaCl, and 0.2 M HCl. The antimicrobial activities for 7, 14, 21, and 28 days against Candida albicans, Staphylococcus aureus, and Streptococcus mutans were assessed according to the JIS Z 2801/ISO 22196 standard. An oral mucosa irritation test was conducted using cheek pouches in five male hamsters according to the ISO 10993-10:2010 standard. The amount of CPC released each day and the cumulative amount released over 28 days (6.12 mg) were less than the daily safe maximum of sore throat medicines (8 mg). Additionally, TC with CPC-Mont could sustain antimicrobial activity against adherent bacteria for 14 days and has no oral mucosa irritation potential.

Antibacterial Effect of Amino Acid-Silver Complex Loaded Montmorillonite Incorporated in Dental Acrylic Resin.

Several dental materials contain silver for antibacterial effect, however the effect is relatively low. The reason for the lower antibacterial efficacy of silver is considered to be the fact that silver ions bind to chloride ions in saliva. To develop new effective silver antibacterial agents that can be useful in the mouth, we synthesized two novel amino acid (methionine or histidine)-silver complexes (Met or His-Ag) loaded with montmorillonite (Mont) and analyzed their antibacterial efficacy. At first the complexes were characterized using nuclear magnetic resonance (NMR), and amino acid-Ag complex-loaded Mont (amino acid-Ag-Mont) were characterized using X-ray diffraction (XRD) and scanning electron microscopy (SEM). The antibacterial efficacy of these materials in dental acrylic resin was then investigated by bacterial growth measurement using a spectrophotometer. As controls, commercially available silver-loaded zeolite and silver-zirconium phosphate were also tested. Dental acrylic resin incorporating His-Ag-Mont strongly inhibited Streptococcus mutans growth. This was explained by the fact that His-Ag complex revealed the highest amounts of silver ions in the presence of chloride. The structure of the amino acid-Ag complexes affected the silver ion presence in chloride and the antibacterial efficacy. His-Ag-Mont might be used as antibacterial agents for dental materials.

Ammonium-to-sodium ion-exchange process at the interlayer of octacalcium phosphate.

Octacalcium phosphate (OCP) has been considered as the layer component of calcium phosphate, but whether it achieves the ionic-exchange ability of conventional layer components is unclear. As OCP is highly biocompatible, understanding its ionic-exchange properties would potentially expand its pharmaceutical and medical applications. Herein, we demonstrate that the substituted cations in ammonium (NH4)-substituted octacalcium phosphate (OCP-NH4) and sodium (Na)-containing ammonium phosphate solutions undergo ion exchanges with OCP interlayers. Replacing NH4 + with Na+ did not alter the crystal structure of OCP, confirming that a substituted cation exchange process similar to that in other layered compounds occurs in OCP.

Aesthetic Silver-Doped Octacalcium Phosphate Powders Exhibiting Both Contact Antibacterial Ability and Low Cytotoxicity.

Since the introduction of biomaterials, infection has been a serious problem in clinical operations. Although several studies have introduced hybrid materials of calcium phosphate and Ag0 nanoparticles (NPs) that exhibit antibacterial activity, released Ag+ ions and Ag0 NPs are highly cytotoxic and the materials require complex fabrication techniques such as laser irradiation. In this study, we introduce a simple one-pot synthesis method based on crystal-engineering techniques to prepare Ag+-substituted octacalcium phosphate (OCP-Ag) powder that simultaneously exhibits antibacterial activity, little change in color, and low cytotoxicity, thereby overcoming the shortcomings of calcium phosphate as a biomaterial. We used AgNO3-containing (NH4)2HPO4 aqueous solutions as reaction solutions in which Ag+ forms soluble complex [Ag(NH3)2]+ ions that are stable at Ag+ concentrations less than ∼30 mmol/L. Hydrolysis of soluble calcium phosphate in this solution led to pure OCP-Ag when the Ag+ concentration was less than ∼30 mmol/L. Crystallographic analysis showed that Ag+ substituted at the P5 PO4-conjugated sites and was uniformly distributed. When the concentration of Ag+ in the reaction solution was varied, the Ag+ content of the OCP-Ag could be controlled. The obtained OCP-Ag exhibited little color change or Ag+ release when immersed in various media; however, it exhibited contact antibacterial ability toward resident oral bacteria. The prepared OCP-Ag showed no substantial cytotoxicity toward undifferentiated and differentiated MC3T3-E1 cells in assays. Notably, when the Ag+ content in OCP-Ag was optimized (Ag: ∼1 at %), it simultaneously exhibited contact antibacterial ability, little color change, and low cytotoxicity.

Development of tissue conditioner containing cetylpyridinium chloride montmorillonite as new antimicrobial agent: Pilot study on antimicrobial activity and biocompatibility.

PURPOSE: The mechanical properties, antimicrobial activity, and biocompatibility of a novel antimicrobial tissue conditioner containing cetylpyridinium chloride with montmorillonite (CPC-Mont) were evaluated. METHODS: To examine the mechanical properties of the novel material, hardness, consistency, and penetration tests were conducted. Antimicrobial activity against Candida albicans (C. albicans) and Staphylococcus aureus (S. aureus) was evaluated. Cell viabilities of fibroblasts and epithelial cells using eluates from materials were measured to evaluate cytotoxicity. In addition, to assess tissue response, animal experiments were conducted. RESULTS: The hardness test results were similar to those of other commercially available materials. The novel tissue conditioner showed good antimicrobial activity against C. albicans and S. aureus compared with other materials. This effect was sustained for a week for C. albicans. In the case of S. aureus, microbial growth was suppressed for up to 3 weeks. Cell viability of the novel material for the eluate at 1 day was significantly less than those of other material for both cells. However, the cell viability at 7 days showed no significant difference. Animal experiments demonstrated that inflammatory responses around materials were not observed on the oral mucosa as other material. CONCLUSION: Within the limitations of this in vitro and in vivo study, the results suggest that the newly developed tissue conditioner containing CPC-Mont has not only excellent antimicrobial properties, but also the same mechanical properties and biocompatibility as tissue conditioners on the market.

Rechargeable anti-microbial adhesive formulation containing cetylpyridinium chloride montmorillonite.

Long-term anti-bacterial effect is a desired ability of any dental material in combating tooth caries as one of the most common and widespread persistent diseases today. Among several cationic quaternary ammonium compounds with antiseptic properties, cetylpyridinium chloride (CPC) is often used in mouthrinses and toothpastes. In this study, we incorporated CPC in a soft phyllosilicate mineral (clay), referred to as montmorillonite (Mont), to enable gradual CPC release with rechargeability. Besides measuring CPC release and recharge, we examined the anti-bacterial effect, cytotoxicity and bonding effectiveness of five experimental adhesive formulations, prepared by adding 1 and 3 wt% CPC_Mont, 3 wt% Mont (without CPC), and 1 and 3 wt% CPC (without Mont) to the commercial adhesive Clearfil S3 Bond ND Quick ('C-S3B'; Kuraray Noritake). Strong inhibition of Streptococcus mutans biofilm formation by CPC_Mont adhesives was confirmed by optical density and SEM. CPC release from CPC_Mont adhesives was higher and lasted longer than from CPC adhesives, while CPC_Mont adhesives could also be recharged with CPC upon immersion in 2 wt% CPC. In conclusion, CPC_Mont technology rendered adhesives anti-bacterial properties with recharge ability, this without reducing its bonding potential, neither increasing its cytotoxicity. STATEMENT OF SIGNIFICANCE: Dental caries is one of the most prevalent chronic diseases in the population worldwide and is the major cause of tooth loss. In this study, we developed cetylpyridinium chloride (CPC) loaded montmorillonite (CPC-Mont) with a long-term antibacterial efficacy to prevent caries. CPC is an antibacterial agent approved by FDA, used as an OTC drug and contained in oral hygiene aids. CPC-Mont was incorporated in a dental adhesive to gradually release CPC. CPC_Mont technology rendered adhesives anti-bacterial properties with rechargeability, this without reducing its bonding potential, neither increasing its cytotoxicity.

Sodium inhibits the formation of ammonium-substituted solid solutions of octacalcium phosphate by filling its substitution site.

Octacalcium phosphate (OCP) is a layered type of calcium phosphate that shows promise for pharmaceutical and biomedical applications because it offers both excellent biocompatibility and a unique, robust crystal structure that readily accepts substitution by various molecules. Although several cations can be incorporated into the OCP crystal lattice by ionic substitution, little is known about the relative probability of different ions to substitute into the OCP crystal lattice. In this study, we focus on Na and NH4, which are known to enter the OCP crystal lattice by ionic substitution. We investigate which of these two is most likely to substitute into OCP in a system containing both ions. In this work, OCP is fabricated from monocalcium phosphate monohydrate via hydrolysis in a solution containing 1 mol L-1 (NH4)2HPO4 with NaCl concentrations ranging from 0 to 100 mmol L-1. X-ray diffraction and thermal analyses indicate that the structure characteristic of NH4 ionic substitution in OCP is attenuated as the Na concentration increases. Furthermore, when the Na concentration exceeds 50 mmol L-1 (Na/NH4 ≥ 1/20), the structure characteristic of NH4 ionic substitution into OCP almost completely disappears. These results indicate that Na is more likely to be ionically substituted into the OCP crystal structure than NH4 and thereby inhibits the structural change of OCP caused by ionic substitution of NH4.

Effectiveness and stability of silane coupling agent incorporated in 'universal' adhesives.

OBJECTIVE: For bonding indirect restorations, some 'universal' adhesives incorporate a silane coupling agent to chemically bond to glass-rich ceramics so that a separate ceramic primer is claimed to be no longer needed. With this work, we investigated the effectiveness/stability of the silane coupling function of the silanecontaining experimentally prepared adhesives and Scotchbond Universal (3MESPE). METHODS AND MATERIALS: Experimental adhesives consisted of Scotchbond Universal and the silane-free Clearfil S3 ND Quick (Kuraray Noritake) mixed with Clearfil Porcelain Bond Activator (Kuraray Noritake) and the two adhesives to which γ-methacryloxypropyltrimethoxysilane (γ-MPTS) was added. Shear bond strength was measured onto silica-glass plates; the adhesive formulations were analyzed using fourier transform infrared spectroscopy (FTIR) and 13C nuclear magnetic resonance (NMR). In addition, shear bond strength onto CAD-CAM composite blocks was measured without and after thermo-cycling ageing. RESULTS: A significantly higher bond strength was recorded when Clearfil Porcelain Bond Activator was freshly mixed with the adhesive. Likewise, the experimental adhesives, to which γ-MPTS was added, revealed a significantly higher bond strength, but only when the adhesive was applied immediately after mixing; delayed application resulted in a significantly lower bond strength. FTIR and (13)C NMR revealed hydrolysis and dehydration condensation to progress with the time after γ-MPTS was mixed with the two adhesives. After thermo-cycling, the bond strength onto CAD-CAM composite blocks remained stable only for the two adhesives with which Clearfil Porcelain Bond Activator was mixed. SIGNIFICANCE: Only the silane coupling effect of freshly prepared silanecontaining adhesives was effective. Clinically, the use of a separate silane primer or silane freshly mixed with the adhesive remains recommended to bond glass-rich ceramics.

Intracellular accumulation of indium ions released from nanoparticles induces oxidative stress, proinflammatory response and DNA damage.

Due to the widespread use of indium tin oxide (ITO), it is important to investigate its effect on human health. In this study, we evaluated the cellular effects of ITO nanoparticles (NPs), indium chloride (InCl3) and tin chloride (SnCl3) using human lung epithelial A549 cells. Transmission electron microscopy and inductively coupled plasma mass spectrometry were employed to study cellular ITO NP uptake. Interestingly, greater uptake of ITO NPs was observed, as compared with soluble salts. ITO NP species released could be divided into two types: 'indium release ITO' or 'tin release ITO'. We incubated A549 cells with indium release ITO, tin release ITO, InCl3 or SnCl2 and investigated oxidative stress, proinflammatory response, cytotoxicity and DNA damage. We found that intracellular reactive oxygen species were increased in cells incubated with indium release ITO, but not tin release ITO, InCl3 or SnCl2. Messenger RNA and protein levels of the inflammatory marker, interleukin-8, also increased following exposure to indium release ITO. Furthermore, the alkaline comet assay revealed that intracellular accumulation of indium ions induced DNA damage. Our results demonstrate that the accumulation of ionic indium, but not ionic tin, from ITO NPs in the intracellular matrix has extensive cellular effects.

In vitro evaluation of the cellular effect of indium tin oxide nanoparticles using the human lung adenocarcinoma A549 cells.

Indium tin oxide (ITO) is widely used in liquid crystal displays (LCDs) or plasma and mobile phone displays. Elevated production and usage of ITO in such displays have led to increased concerns over the safety of industrial workers exposed to particulate aerosols produced during cutting, grinding and polishing of these materials. However, the cellular effects of ITO nanoparticles (NPs) are still unclear, although it has been reported that micro-scale ITO particles induce cytotoxicity. The aim of this study was to examine the potential of ITO NPs to induce cytotoxicity, oxidative stress, and DNA damage using human lung adenocarcinoma A549 cells. Here, stable dispersions of a medium containing ITO NPs were obtained using pre-adsorption and centrifugal fractionation methods, and the A549 cells were incubated in this medium. The ITO NPs showed low cytotoxic effects as shown by the WST-1 and LDH assays. Transmission electron microscopy observations showed the cellular uptake of ITO NPs. The ITO NPs increased the intracellular level of reactive oxygen species and the expression of the heme oxygenase 1 gene. Further, the results of alkaline comet assays showed that ITO NPs induced DNA damage. Thus, these results suggest that ITO NPs possess a genotoxic potential on human lung adenocarcinoma A549 cells.

New approach to a practical quartz crystal microbalance sensor utilizing an inkjet printing system.

The present work demonstrates a valuable approach to developing quartz crystal microbalance (QCM) sensor units inexpensively for reliable determination of analytes. This QCM sensor unit is constructed by inkjet printing equipment utilizing background noise removal techniques. Inkjet printing equipment was chosen as an alternative to an injection pump in conventional flow-mode systems to facilitate the commercial applicability of these practical devices. The results demonstrate minimization of fluctuations from external influences, determination of antigen-antibody interactions in an inkjet deposition, and quantification of C-reactive protein in the range of 50-1000 ng∙mL-1. We thus demonstrate a marketable application of an inexpensive and easily available QCM sensor system.

Lithium recovery from salt lake brine by H2TiO3.

The details of the ion exchange properties of layered H2TiO3, derived from the layered Li2TiO3 precursor upon treatment with HCl solution, with lithium ions in the salt lake brine (collected from Salar de Uyuni, Bolivia) are reported. The lithium adsorption rate is slow, requiring 1 d to attain equilibrium at room temperature. The adsorption of lithium ions by H2TiO3 follows the Langmuir model with an adsorptive capacity of 32.6 mg g(-1) (4.7 mmol g(-1)) at pH 6.5 from the brine containing NaHCO3 (NaHCO3 added to control the pH). The total amount of sodium, potassium, magnesium and calcium adsorbed from the brine was <0.30 mmol g(-1). The H2TiO3 was found capable of efficiently adsorbing lithium ions from the brine containing competitive cations such as sodium, potassium, magnesium and calcium in extremely large excess. The results indicate that the selectivity order Li(+) ≫ Na(+), K(+), Mg(2+), Ca(2+) originates from a size effect. The H2TiO3 can be regenerated and reused for lithium exchange in the brine with an exchange capacity very similar to the original H2TiO3.

Fe-Al layered double hydroxides in bromate reduction: Synthesis and reactivity.

This study presents a rare use of layered double hydroxides of Fe(II) and Al(III) (Fe-Al LDH), as reported for the first time for bromate removal from aqueous solutions. The Fe-Al LDH samples were prepared with Fe/Al molar ratios of 1-4 using a co-precipitation method at pH 7, with subsequent hydrothermal treatment at 120°C. The Fe-Al LDH (molar ratio of Fe/Al=1, 2) with a layered structure exhibited nearly complete removal of bromate from initial concentration of 100µmol/dm(3) at a wide pH range of 4.0-10.5 over a 2h reaction period; the residual bromate concentration in the solution was lower than the detection limit of 0.07µmol/dm(3) (9µg-BrO(3)(-)/dm(3)). During the reaction period, bromide was released into the solution via a reduction process. Reactivity of Fe-Al LDH with a Fe/Al molar ratio of 2 did not decrease the bromate reduction efficiency during 30days.

Synthesis of a novel layered double hydroxides [MgAl(4)(OH)(12)](Cl)(2)·2.4H(2)O and its anion-exchange properties.

A novel layered double hydroxide of Mg and Al with composition [Mg(0.96)Al(4.00)(OH)(12)]Cl(1.86)(CO(3))(0.03)·2.4H(2)O, designated as MgAl(4)-Cl, was synthesized by mixing crystalline gibbsite (γ-Al(OH)(3)) and solid MgCl(2)·6H(2)O with subsequent hydrothermal treatment at 160 °C for 72h. The MgAl(4)-Cl exhibited a crystalline material of a layered structure, as evidenced from X-ray diffraction. Anion uptake experiments with the MgAl(4)-Cl showed that Cl(-) in the interlayer space can be exchanged with anions such as Br(-), H(2)PO(4)(-), CO(3)(2-) or dodecyl sulfate (DS(-)) from aqueous solutions with preservation of the layered structure. Uptake of NO(3)(-), BrO(3)(-) or SO(4)(2-) on the MgAl(4)-Cl showed different behavior; these anions can be exchanged within 1h maintaining the layered structure, but a release of Mg(2+) cations from the sample was observed with increased reaction time, resulting in collapse of the layered structure and formation of the gibbsite phase, as determined from chemical analyses and X-ray diffraction.

Uptake properties of phosphate on a novel Zr-modified MgFe-LDH(CO(3)).

We prepared a novel Zr-modified MgFe-LDH(CO(3)) composite by adding a mixed solution of MgCl(2), FeCl(3), and ZrOCl(2) and another mixed solution of 1mol/dm(3) NaOH and 1mol/dm(3) Na(2)CO(3) to distilled water at a constant pH of 10. The composite exhibited only a poorly crystalline structure, resembling that of layered double hydroxides (LDH) from X-ray diffraction. The phosphate uptake is dependent on pH, decreasing with an increase in pH. This composite shows a much greater uptake of phosphate ions in P-enriched seawater (0.33mg-P/dm(3)) than amorphous zirconium oxide and MgFe-LDH(CO(3)). The uptake isotherm was fitted with a Freundlich relation. These phosphate-uptake behaviors closely resemble those of the relevant Zr-MgAl-LDH, which is estimated to be a composite of MgAl-LDH with amorphous zirconium hydroxide on the surface from X-ray absorption spectroscopy. Therefore, a similar structure of Zr-modified MgFe-LDH(CO(3)) composite probably causes the marked increase in phosphate uptake from P-enriched seawater.

Photoinduced antiviral carbon nanohorns.

Nanocarbons, such as carbon nanohorns (CNH) and carbon nanotubes, are materials of interest in many fields of science and technology because of their remarkable physical properties. We report here a novel approach for using NIR laser-driven CNH as an antiviral agent. NIR laser-driven functional CNH complexes could open the way to a new range of antiviral materials.

Studies on the formation of todorokite-type manganese oxide with different crystalline birnessites by Mg2+-templating reaction.

The formation of todorokite-type manganese oxide (TodMO) by hydrothermal soft chemical reaction (Mg2+ exchange followed by hydrothermal treatment) was studied using three kinds of Na-type birnessite (Na-BirMO) with different crystallinities. Buserite (BusMO) formation by Mg2+ exchange and TodMO formation by hydrothermal treatment progressed in a similar manner regardless of the crystallinity of the initial Na-BirMO, but the crystallinity of the synthesized TodMO depended on that of the initial Na-BirMO. Particle morphology of the synthesized TodMO was related to the crystallinity of the initial Na-BirMO. The amounts of fibrous morphology were gradually increasing, accompanied by increased crystallinity of the initial Na-BirMO. TodMO with a neat fibrous morphology was obtained using a high crystalline Na-BirMO as precursor. Most of the interlayer metal ions (Na+ and Mg2+) were extracted from Na-BirMO and BusMO by treatment with a 0.1 M HCl solution, while only 37% of Mg2+ was extracted from TodMO. Both the acid treatment and the particle morphology may be the most convenient method to distinguish TodMO from BusMO.

Temperature-sensitive photoluminescence of CdSe quantum dot clusters.

The formation of narrow size dispersed and nanometer size aggregates (clusters) of cadmium selenide (CdSe) quantum dots (QDs) and their temperature-sensitive photoluminescence (PL) spectral properties close to room temperature (298 K) are discussed. CdSe QDs formed stable clusters with an average diameter of approximately 27 nm in the absence of coordinating solvents. Using transmission electron microscopy (TEM) imaging, we identified the association of individual QDs with 2-5 nm diameters into clusters of uniform size. A suspension of these clusters in different solvents exhibited reversible PL intensity changes and PL spectral shifts which were correlated with temperature. Although the PL intensity of CdSe QDs encapsulated in host matrixes and the solid state showed a response to temperature under cryogenic conditions, the current work identified for the first time QD clusters showing temperature-sensitive PL intensity variations and spectral shifts at moderate temperatures above room temperature. Temperature-sensitive reversible PL changes of clusters are discussed with respect to reversible thermal trapping of electrons at inter-QD interfaces and dipole-dipole interactions in clusters. Reversible luminescence intensity variations and spectral shifts of QD clusters show the potential for developing sensors based on QD nanoscale assemblies.

Subsecond luminescence intensity fluctuations of single CdSe quantum dots.

Photoluminescence (PL) intermittency characteristics are examined for single quantum dots (QDs) in a CdSe QD sample synthesized at a slow rate at 75 degrees C. Although the PL quantum efficiency was relatively low ( approximately 0.25), we noticed that the PL intensity of single CdSe QDs fluctuated on a subsecond time scale with short-lived "on" and "off" states. The subsecond PL intensity fluctuations of CdSe QDs are different from "on" and "off" PL blinking generally observed for QDs fluctuating on a millisecond to minute time scale. We characterized single QDs by identifying polarized excitations, topographic imaging using atomic force microscopy (AFM), and transmission electron microscopy (TEM). From analysis of the PL intensity trajectories from >100 single CdSe QDs, the average intermittency time was 213 ms. From the PL quantum efficiency, slow growth of QDs, intensity trajectory analyses, and previous reports relating surface trap states and PL properties of QDs, we attribute the subsecond PL intensity fluctuations of single CdSe QDs and short-lived "on" and "off" states to a high-density distribution of homogeneous surface trap states.