Antibacterial and cytotoxic effects of photoexcited Au clusters via blue high-power or white low-power light emitting diode irradiation

The development of photosensitizers and light sources has enabled the use of antimicrobial photodynamic therapy (aPDT) in various dental therapies. In the present study, we compared the antibacterial and cytotoxic effects of Au clusters photoexcited by blue and white LED irradiation. We fabricated novel photosensitizers, captopril-protected gold (Capt-Au) clusters and lysozyme-stabilized gold (Lyz-Au) clusters, for aPDT. Au clusters were then photoexcited by two kinds of light sources, blue high-power and white low-power light-emitting diodes (LEDs). Since white LED contains a wide spectrum of light (400–750 nm), white LED would be relevant for aPDT even if using a low-power source. The turbidity and viability of Streptococcus mutans were assessed following application of Capt-Au clusters (500 μg/mL) or Lyz-Au clusters (1,000 μg/mL) photoexcited by a blue high-power LED (1,000 mW/cm2) or white low-power LED (80 mW/cm2). In addition, the cytotoxicity of Au clusters and LED irradiation was evaluated in NIH3T3 and MC3T3-E1 cells. Au clusters photoexcited by the white low-power LED equally decreased the turbidity and viability of S. mutans compared with blue high-power LED. However, Au clusters photoexcited by white LED irradiation caused decreased cytotoxicity in mammalian cells compared with those photoexcited by blue LED irradiation. In conclusion, white LEDs possess biosafe properties for aPDT using Au clusters

Technique for simple apatite coating on a dental resin composite with light-curing through a micro-rough apatite layer

Tooth root surfaces restored with dental resin composites exhibit inferior biocompatibility. The objective of this study was to develop a simple technique for coating apatite onto a resin composite to improve its surface biocompatibility. First, we fabricated a polymer film coated with a micro-rough apatite layer and pressed it (coating-side down) onto a viscous resin composite precursor. As a result of light-induced curing of the precursor through the overlaid film, the micro-rough apatite layer was integrated with the resin composite and, thus, transferred from the polymer film surface to the cured resin composite surface as a result of the difference in interfacial adhesion strength. The transferred apatite layer attached directly to the cured resin composite without any gaps at the microscopic level. The adhesion between the apatite layer and the cured resin composite was so strong that the layer was not peeled off even by a tape-detaching test. The flexural strength of the resulting apatite-coated resin composite was comparable to that of the clinically used resin composite while satisfying the ISO requirement for dental polymer-based restorative materials. Furthermore, the apatite-coated resin composite showed better cell compatibility than the uncoated resin composite. The present apatite coating technique is well suited for dental treatment because the coating is applied during a conventional light curing procedure through simple utilization of the apatite-coated polymer film in place of an uncoated film. The proposed technique represents a practical evolution in dental treatment using light-curing resin composites, although further in vitro and in vivo studies are needed

チオラート保護金属ナノクラスターの抗菌/光増感作用と抗菌光線力学療法（a-PDT）への応用

Water-soluble Au nanoclusters (NCs) show potential for medical applications. More recently, the Au NCs have been considered as promising photosensitizers for photodynamic therapy (PDT) because of good biocompatibility, photosensitization under nearinfrared light irradiation, good resistance to photobleaching, and target specificity via surface modification. The present article primarily focuses on photosensitization for biomedical therapy applications of Au NCs. The antibacterial action of Ag NCs is also described.出版社サイト巻号ページhttps://www.hyomen.org/vol5no3

Evaluation of antibacterial and cytocompatible properties of multiple-ion releasing zinc-fluoride glass nanoparticles

Zinc-fluoride glass nanoparticles (Zinc-F) release several ions, such as fluoride, zinc and calcium ions, through acid-base reactions. The aim of this study was to evaluate the antibacterial and cytotoxic properties of Zinc-F. Antibacterial tests showed that a Zinc-F eluting solution significantly reduced the turbidity and colony-forming units of Streptococcus mutans and Actinomyces naeslundii, compared to that of calcium-fluoroaluminosilicate glass nanoparticles without zinc ions. In live/dead staining, Zinc-F eluate significantly decreased green-stained bacterial cells, indicating live cells, compared with the control (no application). Human dentin coated with Zinc-F showed suppressed S. mutans and A. naeslundii biofilm formation. Additionally, Zinc-F eluate showed low cytotoxic effects in osteoblastic and fibroblastic cells. Therefore, our findings suggested that Zinc-F exhibits antibacterial and biocompatible properties through multiple-ion release

Antibacterial tooth surface created by laser-assisted pseudo-biomineralization in a supersaturated solution

A technique for implementing biocompatible and antibacterial functions to a targeted region on tooth surfaces has potential in dental treatments. We have recently demonstrated pseudo-biomineralization, i.e., the growth of an apatite layer on a human dentin substrate by a laser-assisted biomimetic (LAB) process, based on pulsed laser irradiation in a supersaturated CaP solution. In this study, pseudo-biomineralization was induced in the presence of fluoride ions using the LAB process in order to fabricate an antibacterial fluoride-incorporated apatite (FAp) layer on the dentin surface. After processing for 30 min, a micron-thick FAp layer was formed heterogeneously at the laser-irradiated solid-liquid interface via pseudo-biomineralization. A time-course study revealed that the LAB process first eliminated the pre-existing organic layer, while allowing fluoride incorporation into the dentin surface within 1 min. Within 5 min, FAp nanocrystals precipitated on the dentin surface. Within 30 min, these nanocrystals acquired a pillar-like structure that was weakly oriented in the direction normal to the substrate surface to form a dense micron-thick layer. This layer was integrated seamlessly with the underlying dentin without any apparent gaps. The FAp layer exhibited antibacterial activity against a major oral bacterium, Streptococcus mutans. The proposed LAB process is expected to be a useful new tool for tooth surface functionalization via facile and area-specific pseudo-biomineralization

Human Dentin Coated with Silver Nanoclusters Exhibits Antibacterial Activity against Streptococcus mutans

Silver nanoclusters (AgNCs) are ultrasmall in size (< 2 nm) and are expected to be an effective antibacterial substance to combat oral infective diseases. In the present study, we synthesized AgNCs (Ag∼75) as the main component for esthetic and antibacterial application against the caries pathogen Streptococcus mutans. The results showed that AgNCs significantly reduced the turbidity and viability of S. mutans. In addition, the bactericidal effects of AgNCs were confirmed by LIVE/DEAD staining. After AgNC application to human dentin, no discoloration of dentin was observed, as compared to silver diamine fluoride application, and AgNC-treated dentin showed an inhibitory effect on colony formation by S. mutans. Therefore, AgNCs appear to be beneficial for dental therapy as an antibacterial and/or esthetic substrate

Photodynamic inactivation of oral bacteria with silver nanoclusters/rose bengal nanocomposite

Antimicrobial photodynamic therapy (a-PDT) is a promising anti-infective technique for generation of singlet oxygen (1O2) to target dental disease. However, conventional organic photosensitizers have problems for clinical use in terms of cytotoxicity, quenching of a-PDT activity by self-dimerization, and the lack of long-term antibacterial effect. We herein propose silver nanoclusters/rose bengal nanocomposite (AgNCs/RB) as a novel photosensitizer with two primary antibacterial effects: (1) 1O2 generation by irradiated RB and (2) Ag+ ion release from AgNCs. AgNCs/RB irradiated with white light-emitting diode (LED) for a short irradiation time of 1 min significantly decreased the bacterial turbidity of Streptococcus mutans, Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans (P < 0.05). In SEM, TEM and LIVE/DEAD staining images, photoexcited AgNCs/RB reduced S. mutans colonization, destroyed the cell membrane, and increased the number of dead cells. The antibacterial efficiency of photoexcited AgNCs/RB was greater than that of AgNCs or RB alone (P < 0.05), suggesting a synergistic effect of 1O2 and Ag+ ions from photoexcited AgNCs/RB. By contrast, photoexcited AgNCs/RB did not affect WST-8 and LDH activities and morphology of NIH3T3 mammalian cells, indicating low cytotoxicity. Interestingly, the antibacterial activity of AgNCs/RB on S. mutans was maintained even after the cessation of LED irradiation, indicating a long-term antibacterial effect due to released Ag+ ions. The present AgNCs/RB photosensitizers provide effective synergistic antibacterial effects for dental a-PDT via 1O2 and Ag+ ions coupled with low cytotoxicity

Fluoridated Apatite Coating on Human Dentin via Laser-Assisted Pseudo-Biomineralization with the Aid of a Light-Absorbing Molecule

A simple, area-specific coating technique for fluoridated apatite (FAp) on teeth would be useful in dental applications. Recently, we achieved area-specific FAp coating on a human dentin substrate within 30 min by a laser-assisted biomimetic (LAB) process; pulsed Nd:YAG laser irradiation in a fluoride-containing supersaturated calcium phosphate solution (FCP solution). The LAB-processed, FAp-coated dentin substrate exhibited antibacterial activity against a major oral bacterium, Streptococcus mutans. In the present study, we refined the LAB process with a combination of a dental diode laser and a clinically approved light-absorbing molecule, indocyanine green (ICG). A micron-thick FAp layer was successfully formed on the dentin surface within only 3 min by the refined LAB process, i.e., dental diode laser irradiation in the FCP solution following ICG treatment. The ICG layer precoated on the dentin substrate played a crucial role in inducing rapid pseudo-biomineralization (FAp layer formation) on the dentin surface by absorbing laser light at the solid-liquid interface. In the refined LAB process, the precoated ICG layer was eliminated and replaced with the newly formed FAp layer composed of vertically oriented pillar-like nanocrystals. Cross-sectional ultrastructural analysis revealed a smooth interface between the FAp layer and the dentin substrate. The refined LAB process has potential as a tool for the tooth surface functionalization and hence, is worth further process refinement and in vitro and in vivo studies