Direct Quantification of Attogram Levels of Strontium-90 in Microscale Biosamples Using Isotope Dilution-Thermal Ionization Mass Spectrometry Assisted by Quadrupole Energy Filtering.

Although thermal ionization mass spectrometry (TIMS) has been employed for the high-precision analysis of isotope ratios, direct quantification of artificial mono-nuclide in the environment is difficult by even using isotope dilution (ID) due to the coexistence of the great magnitude of natural stable nuclides or isobars. In traditional TIMS and ID-TIMS, a sufficient amount of stable Sr doped on a filament is required to realize a stable and adequate ion-beam intensity (i.e., thermally ionized beams). However, the background noise (BGN) at m/z 90, detected by an electron multiplier, disturbs 90Sr analysis at low concentration levels due to peak tailing of a significant 88Sr ion beam dependent on the 88Sr-doping amount. Here, TIMS assisted by quadruple energy filtering was successfully employed for the direct quantification of attogram levels of an artificial monoisotopic radionuclide strontium-90 (90Sr) in microscale biosamples. Direct quantification was achieved by integrating the ID quantification of natural Sr and simultaneous 90Sr/86Sr isotope ratio analysis. Additionally, the measurement amount calculated by the combination of the ID and intercalibration was corrected for the net result amount of 90Sr by subtracting dark noise and the detected amount derived from the survived 88Sr, which are equivalent with the BGN intensity at m/z 90. Background correction revealed that the detection limits were in the range of 6.15 × 10-2-3.90 × 10-1 ag (0.31-1.95 µBq), depending on the concentration of natural Sr in a 1 µL sample, and the quantification of 0.98 ag (5.0 µBq) of 90Sr in 0-300 mg/L of natural Sr was successful. This method could analyze small sample quantities (1 µL), and the quantitative results were verified against authorized radiometric analysis techniques. Furthermore, the amount of 90Sr in actual teeth was successfully quantified. This method will be a powerful tool for measuring 90Sr in the measurement of micro-samples, which are required to assess and understand the degree of internal radiation exposure.

Shear bond strength of an acrylic resin to a ceria-stabilized zirconia-alumina nanocomposite.

STATEMENT OF PROBLEM: Ceria-stabilized zirconia-alumina nanocomposite (Ce-TZP-Al2O3) has properties that may be suitable for partial denture frameworks. However, studies on its adhesion strength and durability with denture base resin are lacking. PURPOSE: The purpose of this in vitro study was to determine the optimal surface treatment for Ce-TZP-Al2O3 to secure a durable bond with an acrylic resin. MATERIAL AND METHODS: The surface of Ce-TZP-Al2O3 test specimens was alumina airborne-particle abraded (Group APA) and then treated with 10-methacryloyloxydecyl dihydrogen phosphate (MDP) (Group MDP) and 2 silica coating methods: the flame spraying method (Group SLP) and the tribochemical treatment (110 µm: Group TRB-P, 30 µm: Group TRB-S). TRB-P and TBR-S were further treated by MDP (Group CBT-P and CBT-S). Autopolymerizing acrylic resin was bonded to the specimens, and the shear bond strength was tested after thermocycling (5 °C and 60 °C, 10 000 cycles). The area of the resin remaining on the fractured surfaces was also measured. To evaluate the effect of the surface treatment condition on shear bond strength and the resin remaining, 1-way analysis of variance (ANOVA) was conducted, followed by the Tukey multiple comparison post hoc test. Additionally, the effect of thermocycling on the specimens was evaluated by the Student t test. RESULTS: After placement in deionized water for 24 hours, the shear bond strengths of Group MDP and 2 types of combination treatment (Groups CBT-P and CBT-S) were significantly higher than those of Groups SLP, TRB-P, and TRB-S (P<.05). Moreover, the fractured surface of all the treatment conditions except Group APA showed cohesive failure. The shear bond strength as a result of all treatment conditions decreased significantly after thermocycling (P<.05). Group CBT-S showed the highest shear bond strength; however, no significant differences were found between Groups CBT-S and MDP (P=.908). In particular, the area of resin remaining on the fractured surfaces of Group CBT-S was 100% (cohesive failure). CONCLUSIONS: The combined surface treatment of alumina airborne-particle abrasion and tribochemical treatment, along with primer treatment using silane coupling and an MDP monomer, improved the adhesion strength and adhesion durability between base resins and Ce-TZP-Al2O3.

Intranasal vaccination with HBs and HBc protein combined with carboxyl vinyl polymer induces strong neutralizing antibody, anti-HBs IgA, and IFNG response.

Hepatitis B virus (HBV) infection causes acute and chronic hepatitis, which is a major public health concern worldwide. Immunization methods incorporating hepatitis B surface-small (HBs-S) antigen and hepatitis B core antigen (HBc) have been proposed as candidate therapeutic vaccines, but the elimination of existing HBV infection remains a challenge. To enhance the efficacy of HBs and HBc vaccination, we investigated HBs-large (HBs-L) as an immunogen, and carboxyl vinyl polymer (CVP) as an excipient. HBs-S or HBs-L, in combination with HBc antigen, was administered subcutaneously (without CVP) or intranasally (with or without CVP) for the evaluation of immune response in the tree shrew, which is considered to be a suitable small animal model of HBV infection. Immunization with HBs-L antigen by either route induced a rapid IgG response. Intranasal immunization with HBs-S or HBs-L and HBc formulated with CVP strongly induced neutralizing antibody activity, IgA response, and HBc-specific expression of the interferon gamma-encoding gene. These data indicated the potential of HBs-L and HBc intranasal immunization with CVP, not only as a therapeutic vaccine, but also as a prophylactic vaccine candidate.

Streptococcus sanguinis Noncoding cia-Dependent Small RNAs Negatively Regulate Expression of Type IV Pilus Retraction ATPase PilT and Biofilm Formation.

Small noncoding RNAs (sRNAs) have been identified as important regulators of gene expression in various cellular processes. cia-dependent small RNAs (csRNAs), a group of sRNAs that are controlled by the two-component regulatory system CiaRH, are widely conserved in streptococci, but their targets have been identified only in Streptococcus pneumoniaeStreptococcus sanguinis, a pioneer colonizer of teeth and one of the most predominant bacteria in the early oral biofilm, has been shown to have six csRNAs. Using computational target prediction and the luciferase reporter assay, we identified pilT, a constituent of the type IV pilus operon, as a negative regulatory target for one of the csRNAs, namely, csRNA1-1, in S. sanguinis RNA-RNA electrophoretic mobility shift assay using a nucleotide exchange mutant of csRNA1-1 revealed that csRNA1-1 binds directly to pilT mRNA. In addition, csRNA1-1 and csRNA1-2, a putative gene duplication product of csRNA1-1 that is tandemly located in the S. sanguinis genome, negatively regulated S. sanguinis biofilm formation. These results suggest the involvement of csRNAs in the colonization step of S. sanguinis.

Potential effect of cationic liposomes on interactions with oral bacterial cells and biofilms.

CONTEXT: Although oral infectious diseases have been attributed to bacteria, drug treatments remain ineffective because bacteria and their products exist as biofilms. Cationic liposomes have been suggested to electrostatically interact with the negative charge on the bacterial surface, thereby improving the effects of conventional drug therapies. However, the electrostatic interaction between oral bacteria and cationic liposomes has not yet been examined in detail. OBJECTIVE: The aim of the present study was to examine the behavior of cationic liposomes and Streptococcus mutans in planktonic cells and biofilms. MATERIALS AND METHODS: Liposomes with or without cationic lipid were prepared using a reverse-phase evaporation method. The zeta potentials of conventional liposomes (without cationic lipid) and cationic liposomes were -13 and 8 mV, respectively, and both had a mean particle size of approximately 180 nm. We first assessed the interaction between liposomes and planktonic bacterial cells with a flow cytometer. We then used a surface plasmon resonance method to examine the binding of liposomes to biofilms. We confirmed the binding behavior of liposomes with biofilms using confocal laser scanning microscopy. RESULTS: The interactions between cationic liposomes and S. mutans cells and biofilms were stronger than those of conventional liposomes. Microscopic observations revealed that many cationic liposomes interacted with the bacterial mass and penetrated the deep layers of biofilms. DISCUSSION AND CONCLUSION: In this study, we demonstrated that cationic liposomes had higher affinity not only to oral bacterial cells, but also biofilms than conventional liposomes. This electrostatic interaction may be useful as a potential drug delivery system to biofilms.

Nanomechanical properties and molecular structures of in vitro mineralized tissues on anodically-oxidized titanium surfaces.

The biomechanical stability of mineralized tissues at the interface between implant surface and bone tissue is of critical importance. Anodically oxidized titanium prepared in a chloride solution results in enhanced mineralization of adherent osteoblasts and has antimicrobial activity against oral microorganisms. We evaluated the nanomechanical properties and molecular structures of the in vitro mineralized tissues developing around anodically oxidized titanium surfaces with and without preparation in chloride solution. Anodically oxidized titanium surfaces showed superior osteogenic gene expressions than those of thermally oxidized and bare titanium surfaces. Preparation of anodically oxidized titanium in chloride enhanced the production of mineralized tissue around it. However, the mineralized tissue around anodically oxidized titanium prepared without chloride had increased mineral:matrix and cross-linking ratios, resulting in higher hardness and lower elasticity. FROM THE CLINICAL EDITOR: In this study anodically oxidized titanium was used to enhance the biomechanical stability of mineralized tissues at the implant surface -- bone tissue interface. The mineralized tissue around anodically oxidized titanium prepared without chloride had increased mineral:matrix and cross-linking ratios, resulting in higher hardness and lower elasticity.

Biodistribution assessment of cationic pullulan nanogel, a nasal vaccine delivery system, in mice and non-human primates.

Cationic cholesteryl-group-bearing pullulan nanogel (cCHP-nanogel) is an effective drug-delivery system for nasal vaccines. However, cCHP-nanogel-based nasal vaccines might access the central nervous system due to its close proximity via the olfactory bulb in the nasal cavity. Using real-time quantitative tracking of the nanogel-based nasal botulinum neurotoxin and pneumococcal vaccines, we previously confirmed the lack of deposition of vaccine antigen in the cerebrum or olfactory bulbs of mice and non-human primates (NHPs), rhesus macaques. Here, we used positron emission tomography to investigate the biodistribution of the drug-delivery system itself, cCHP-nanogel after mice and NHPs were nasally administered with 18F-labeled cCHP nanogel. The results generated by the PET analysis of rhesus macaques were consistent with the direct counting of radioactivity due to 18F or 111In in dissected mouse tissues. Thus, no depositions of cCHP-nanogel were noted in the cerebrum, olfactory bulbs, or eyes of both species after nasal administration of the radiolabeled cCHP-nanogel compound. Our findings confirm the safe biodistribution of the cCHP-nanogel-based nasal vaccine delivery system in mice and NHPs.

Antimicrobial and osteogenic properties of a hydrophilic-modified nanoscale hydroxyapatite coating on titanium.

Hydroxyapatite (HA)-coated titanium (Ti) is commonly used for implantable medical devices. This study examined in vitro osteoblast gene expression and antimicrobial activity against early and late colonizers of supra-gingival plaque on nanoscale HA-coated Ti prepared by discharge in a physiological buffered solution. The HA-coated Ti surface showed super-hydrophilicity, whereas the densely sintered HA and Ti surfaces alone showed lower hydrophilicity. The sintered HA and HA-coated Ti surfaces enhanced osteoblast phenotypes in comparison with the bare Ti surface. The HA-coated Ti enabled antimicrobial activity against early colonizers of supra-gingival plaques, namely Streptococcus mitis and Streptococcus gordonii. Such antimicrobial activity may be caused by the surface hydrophilicity, thereby leading to a repulsion force between the HA-coated Ti surface and the bacterial cell membranes. On the contrary, the sintered HA sample was susceptible to infection of microorganisms. Thus, hydrophilic-modified HA-coated Ti may have potential for use in implantable medical devices. From the Clinical Editor: This study establishes that Hydroxyapatite (HA)-coated titanium (Ti) surface of implanted devices may result in an optimal microenvironment to control and prevent infections and may have potential future clinical applications.

High-resolution mechanical mapping of the adhesive-dentin interface: The effect of co-monomers in 10-methacryloyloxydecyl dihydrogen phosphate.

The presence of 10-methacryloyloxydecyl dihydrogen phosphate (MDP) at the adhesive-dentin interface enables ionic binding to calcium salts, which results in rigid nano-layering within the submicron scale resin-dentin interdiffusion zone. MDP has been used with additional co-monomers, such as hydroxyethyl methacrylate (HEMA) and/or 4-methacryloyloxyethyl-trimellitic acid (4-MET), mainly to enhance the chemical bonding properties. However, the use of co-monomers may compromise the rigidity of the adhesive-dentin interface. In this study, we use high-resolution mechanical mapping across the interface to discern the in situ mechanical properties of each target region at the nanoscale. Visualization by modulus mapping demonstrated that HEMA increases the diffusion properties of MDP into dentin structures. However, the rigidity of the adhesive-dentin interface indicated by the storage modulus was markedly lower in MDP containing HEMA than in MDP containing 4-MET. Dynamic indentation testing revealed that the bonding layer was more deformable in the presence of HEMA. Moreover, the presence of MDP in the bonding layer might also increase the deformability because the polymerization linearity allows a large degree of viscoelasticity. These factors also diminish the rigidity of the adhesive-dentin interface. Within the limitations of this study, our findings demonstrated that 4-MET is a better co-monomer than HEMA in MDP-based dental adhesives. Modulus mapping and nanoindentation are introduced as new tests for the adhesive-dentin interface to address queries about the effectiveness of dental adhesives.

A nanogel-based trivalent PspA nasal vaccine protects macaques from intratracheal challenge with pneumococci.

Current polysaccharide-based pneumococcal vaccines are effective but not compatible with all serotypes of Streptococcus pneumoniae. We previously developed an adjuvant-free cationic nanogel nasal vaccine containing pneumococcal surface protein A (PspA), which is expressed on the surfaces of all pneumococcal serotypes. Here, to address the sequence diversity of PspA proteins, we formulated a cationic nanogel-based trivalent pneumococcal nasal vaccine and demonstrated the vaccine's immunogenicity and protective efficacy in macaques by using a newly developed nasal spray device applicable to humans. Nasal vaccination of macaques with cationic cholesteryl pullulan nanogel (cCHP)-trivalent PspA vaccine effectively induced PspA-specific IgGs that bound to pneumococcal surfaces and triggered complement C3 deposition. The immunized macaques were protected from pneumococcal intratracheal challenge through both inhibition of lung inflammation and a dramatic reduction in the numbers of bacteria in the lungs. These results demonstrated that the cCHP-trivalent PspA vaccine is an effective candidate vaccine against pneumococcal infections.

Combination treatment of tribochemical treatment and phosphoric acid ester monomer of zirconia ceramics enhances the bonding durability of resin-based luting cements.

The aim of the present study was to evaluate the bonding durability of resin-based luting cement to partially stabilized tetragonal zirconia (Y-TZP) achieved by combination treatment of tribochemical (TBC) treatment and two different phosphate acid ester monomers. Two phosphate acid ester monomers (EP: Epricord opaque primer, AZ: AZ primer) were applied to each surface modification followed by application of resin-based luting cement (Rely-X ARC). Bonding specimens were placed in deionized water at 37 degrees C and stored for 24 h. The other groups were subjected to 30,000 cycles of a thermal stress for the durability test. Shear bond tests were done using a universal testing machine at 1 mm/min. Shear bond strengths of combination treatments using EP and AZ on TBC treatment after thermal stress showed no significant difference (p>0.05) compared with those of storage after 24 h. Combination treatment using phosphoric acid ester monomer could achieve a durable bond.

Transparent dentin region in the tooth root.

Transparent dentin in the tooth root forms during the aging process. The transparent dentin is weaker than normal opaque dentin, which may explain the occurrence of root dentin fractures in aged individuals. Tooth fractures are caused by the brittleness of the transparent dentin. Clinical procedures in aged persons require modification to accommodate the reduced strength of the dentin tissue.

Anisotropic ultimate strength and microscopic fracture patterns during tensile testing in the dentin-enamel junction region.

The dentin-enamel junction (DEJ), which is the interface between the outer tooth enamel and the underlying dentin, has unique biomechanical properties that allow the two different materials to function together without fracture. The aim of this study was to investigate the anisotropic ultimate tensile strength of the DEJ. Twenty dumbbell-shaped samples were prepared from 10 extracted human molar teeth for tensile strength testing. The maximum tensile forces at fracture were measured in the direction parallel (10 samples) and perpendicular (10 samples) to the DEJ. The mean tensile strength of the samples was significantly higher under parallel forces (30.9±3.3 MPa) than under perpendicular forces (20.6±4.8 MPa; p<0.05). Our findings confirm the structural anisotropy of the DEJ.

3D imaging of proximal caries in posterior teeth using optical coherence tomography.

Optical coherence tomography (OCT) can create cross-sectional images of tooth without X-ray exposure. This study aimed to investigate the diagnostic accuracy of 3D imaging of OCT for proximal caries in posterior teeth. Thirty-six human molar teeth with 51 proximal surfaces visibly 6 intact, 16 slightly demineralized, and 29 distinct carious changes were mounted to take digital radiographs and 3D OCT images. The sensitivity, specificity and area under the receiver operating characteristic curve (AUC) for the diagnosis of enamel caries and dentin caries were calculated to quantify the diagnostic ability of 3D OCT in comparison with digital radiography. Diagnostic accuracy was evaluated by the agreement with histology using weighted Kappa. OCT showed significantly higher sensitivity, AUC and Kappa values than radiography. OCT can be a safer option for the diagnosis of proximal caries in posterior teeth that can be applied to the patients without X-ray exposure.

Properties of a gypsum-bonded magnesia investment using a K2SO4 solution for titanium casting.

The purpose of the study was to investigate the effects of a K2SO4 solution on the improvement of the properties of an experimental magnesia-based gypsum-bonded investment. The characteristics of the experimental investment with different ratios of a K2SO4 solution were as follows: setting time ranged from 34 to 152 minutes, green and fired compressive strengths were approximately 2.15 to 5.81 MPa and 1.63 to 2.45 MPa, respectively; thermal expansion was 0.38 to 0.47% at 700 degrees C, which did not show any significant differences due to the concentration of the K2SO4 solution. Titanium casting could be obtained using the experimental investment mixed with a 1% K2SO4 solution, and the reaction layer thickness of the casting was less than 200 microm. The results suggest that the experimental investment mixed with 1% K2SO4 showed acceptable physical properties and casting characteristics for titanium casting.

A review of dental CAD/CAM: current status and future perspectives from 20 years of experience.

In this article, we review the recent history of the development of dental CAD/CAM systems for the fabrication of crowns and fixed partial dentures (FPDs), based on our 20 years of experience in this field. The current status of commercial dental CAD/CAM systems developed around the world is evaluated, with particular focus on the field of ceramic crowns and FPDs. Finally, we discuss the future perspectives applicable to dental CAD/CAM. The use of dental CAD/CAM systems is promising not only in the field of crowns and FPDs but also in other fields of dentistry, even if the contribution is presently limited. CAD/CAM technology will contribute to patients' health and QOL in the aging society.

Comparison of nanohardness between coronal and radicular intertubular dentin.

This study investigated the hardness and Young's modulus of coronal and radicular intertubular dentin. Ten bovine teeth were each divided into coronal and radicular groups, and the flat surfaces of the coronal and radicular dentin were subsequently processed along the tooth axis. The hardness and Young's modulus of the coronal and radicular intertubular dentin were evaluated using nanoindentation tests, at two locations per tooth. Mean hardness and Young's modulus values were statistically compared by one-way ANOVA and Fisher's PLSD test. The hardness of coronal intertubular dentin was 0.81 +/- 0.05 GPa and that of radicular dentin was 0.55 +/- 0.02 GPa. Additionally, the Young's modulus of coronal intertubular dentin was 26.60 +/- 2.19 GPa and that of radicular dentin was 20.89 +/- 1.10 GPa. Findings of this study revealed that the hardness and Young's modulus of coronal intertubular dentin were greater than those of radicular intertubular dentin.

Detection of enamel subsurface lesions by swept-source optical coherence tomography.

This study aimed to non-destructively analyze the progression of subsurface enamel caries using swept-source optical coherence tomography (SS-OCT), a recently developed imaging analysis modality. Artificial enamel caries at various stages of demineralization were created in bovine tooth enamel using a modified lactic acid gel system. Untreated enamel served as a control. OCT images from cross-sections of enamel were collected. Mineral density, distribution, and histological changes were analyzed using an electron probe microanalyzer, scanning electron microscopy, and contact microradiography. The Raman spectrum and X-ray structural analysis of the enamel surfaces were determined. SS-OCT detected significant differences in mineral loss among the samples. A high-brightness image was confirmed, along with changes in the respective brightness graphs proportionate to the degree of demineralization. SS-OCT can potentially be used to evaluate the progression of incipient enamel carious lesions.

Pentosidine correlates with nanomechanical properties of human jaw bone.

Initial intimate apposition between implant fixtures and host bone at the surgical site is a critical factor for osseointegration of dental implants. The advanced glycation end products accumulated in the jaw bone could lead to potential failure of a dental implant during the initial integration stage, because of the inferior bone mechanical property associated with the abnormal collagen cross-linking at the material level. Here, we demonstrate the lowered creep deformation resistance and reduced dimensional recovery of jaw bone in line with high levels of pentosidine accumulation in the bone matrix which likely correlate with the pentosidine level in blood plasma. Peripheral blood samples and cortical bone samples at the surgical site were obtained from patients scheduled for dental implants in the mandible. The pentosidine levels in blood plasma were assessed. Subsequently, the relative pentosidine levels and the mechanical properties of the jaw bone were quantified by Raman microspectroscopy and nanoindentation, respectively. The nanoindentation tests revealed less creep deformation resistance and reduced time-dependent dimensional recovery of bone samples with the increase in the relative pentosidine level in the bone matrix. Higher tan δ values at the various frequencies during the dynamic indentation tests also suggested that viscoelasticity is associated with the relative intensity of pentosidine in the jaw bone matrix. We found a positive correlation between the pentosidine levels in blood plasma and the bone matrix, which in turn reduced the mechanical property of the jaw bone at the material level. Increased creep and reduced dimensional recovery of the jaw bone may diminish the mechanical interlocking of dental implants during the initial integration stage. Given the likely correlation between the plasma pentosidine level and the mechanical properties of bone, measurement of the plasma pentosidine level could serve as a new index to assess jaw bone matrix quality in advance of implant surgery.

Quantitative/qualitative analysis of adhesive-dentin interface in the presence of 10-methacryloyloxydecyl dihydrogen phosphate.

Dental adhesive provides effective retention of filling materials via adhesive-dentin hybridization. The use of co-monomers, such as 10-methacryloyloxydecyl dihydrogen phosphate (10-MDP), is thought to be crucial for hybridization owing to their ionic-binding to calcium and co-polymerization in the polymerizable adhesives. Optimal hybridization partly depends on the mechanical properties of polymerized adhesives, which are likely to be proportional to the degree of conversion ratio. This study assessed the correlation between polymerization quality and mechanical properties at the adhesive-dentin interfaces in the presence or absence of 10-MDP. In situ Raman microspectroscopy and nanoindentation tests were used concurrently to quantify the degree of conversion ratio and dynamic mechanical properties across the adhesive-dentin interfaces. Despite the excellent diffusion and apparent higher degree of co-polymerization, 10-MDP reduced the elastic modulus of the interface. The higher viscoelastic properties of the adhesive are suggestive of poor polymerization, namely polymerization linearity related to the long carboxyl chain of 10-MDP. Such reduced mechanical integrity of hybridization could also be associated with the inhibition of nano-layering between 10-MDP and mineralized tissue in the presence of hydroxyethyl methacrylate (HEMA). This potential drawback of HEMA necessitates further qualitative/quantitative characterization of adhesive-dentin hybridization using a HEMA-free/low concentration experimental 10-MDP monomer, which theoretically possesses superior chemical bonding potential to the current HEMA-rich protocol.