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Objective: To develop a catalytic hairpin assembly (CHA)-based fluorescent assay for the detection of the target RNA of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), so as to realize the rapid nucleic acid testing of SARS-CoV-2. Methods: A 24-nt segment of the SARS-CoV-2 nucleocapsid protein gene (N gene, NC_045512.2) was chosen as the target RNA and the hairpin motif 1 (H1) and hairpin motif 2 (H2) were designed based on the principle of CHA reaction. The H1 motif was labelled with a fluorophore group as well as a quencher group. When the target RNA was added to the hairpin motifs, CHA reaction was triggered at room temperature (25 ℃), which led to the amplification of fluorescence signal, thereby enabling the rapid detection of the target RNA. After the optimization of the hairpin motifs and the experimental conditions, the sensitivity and the specificity of the testing method were measured to evaluate its performance. Results: We successfully constructed a CHA-based fluorescent assay specifically for the target RNA of SARS-CoV-2. With this method, testing could be completed at room temperature within 30 min. This testing method exhibited excellent specificity and could be used to accurately distinguish the perfectly-matched target RNA from the target RNA with single-base mutations. In addition, the testing method demonstrated good sensitivity, with a detection limit of 50 pmol/L. Conclusion: The proposed assay enables the simple and rapid detection of the SARS-CoV-2 target RNA with excellent sensitivity and specificity, showing great promise for further optimization and subsequent clinical application for the rapid detection of SARS-CoV-2 nucleic acid.

Design, Synthesis, and Cellular Imaging Application of a Fluorescent Probe Based on Fluoride Ion-Induced Cyclization of Phenothiazine Derivatives.

Fluoride is both necessary and potentially harmful in excessive amounts, making its detection crucial. Fluorescent probes provide a sensitive and selective means for this purpose. In this study, we developed and synthesized a fluorescent probe for LDT using phenothiazine derivatives and aryl vinyl nitrile. Initially non-fluorescent, the probe undergoes a Si-O bond breakage in the presence of fluoride ions, resulting in the formation of a larger conjugated system and subsequent fluorescence emission. The probe exhibits superior selectivity and sensitivity towards fluoride ions, with a detection limit of 0.35 µM. Moreover, cellular imaging experiments demonstrated the probe's effectiveness in recognizing fluoride ions within HepG2 cells.

The survival of murine hepatitis virus (a surrogate of SARS-CoV-2) on conventional packaging materials under cold chain conditions.

Introduction: The cold chain conditions have been suggested to facilitate long-distance transmission of SARS-CoV-2, but it is unclear how viable the virus is on cold chain packaging materials. Methods: This study used the MHV-JHM strain of murine hepatitis virus as a model organism to investigate the viability of SARS-CoV-2 on foam, plastic, cardboard, and wood sheets at different temperatures (-40°C, -20°C, and 4°C). In addition, the ability of peracetic acid and sodium hypochlorite to eliminate the MHV-JHM on plastic and cardboard sheets were also evaluated. Results: The results indicate that MHV-JHM can survive on foam, plastic, or cardboard sheets for up to 28 days at -40°C and -20°C, and up to 14 days on foam and plastic surfaces at 4°C. Although viral nucleic acids were still detectable after storing at 4°C for 28 days, the corresponding virus titer was below the limit of quantification (LOQ). Discussion: The study highlights that a positive nucleic acid test result may not indicate that the virus is still viable, and confirms that peracetic acid and sodium hypochlorite can effectively eliminate MHV-JHM on packaging materials under cold chain conditions.

Quantifying global colonization pressures of alien vertebrates from wildlife trade.

The global trade in live wildlife elevates the risk of biological invasions by increasing colonization pressure (the number of alien species introduced to an area). Yet, our understanding of species traded as aliens remains limited. We created a comprehensive global database on live terrestrial vertebrate trade and use it to investigate the number of traded alien species, and correlates of establishment richness for aliens. We identify 7,780 species involved in this trade globally. Approximately 85.7% of these species are traded as aliens, and 12.2% of aliens establish populations. Countries with greater trading power, higher incomes, and larger human populations import more alien species. These countries, along with island nations, emerge as hotspots for establishment richness of aliens. Colonization pressure and insularity consistently promote establishment richness across countries, while socio-economic factors impact specific taxa. Governments must prioritize policies to mitigate the release or escape of traded animals and protect global biosecurity.

Pathogenic selection promotes adaptive immune variations against serious bottlenecks in early invasions of bullfrogs.

Adaptive genetic variations are key for understanding evolutionary processes influencing invasions. However, we have limited knowledge on how adaptive genetic diversity in invasive species responds to new pathogenic environments. Here, we compared variations in immune major histocompatibility complex (MHC) class-II ß gene and neutral loci in relation to pathogenic chytrid fungus (Batrachochytrium dendrobatidis, Bd) infection across invasive and native populations of American bullfrog between China and United States (US). Chinese invasive populations show a 60% reduction in neutral cytb variations relative to US native populations, and there were similar MHC variation and functional diversity between them. One MHC allele private to China was under recent positive selection and associated with decreased Bd infection, partly explaining the lower Bd prevalence for Chinese populations than for native US populations. These results suggest that pathogen-mediated selection favors adaptive MHC variations and functional diversity maintenance against serious bottlenecks during the early invasions (within 15 generations) of bullfrogs.

Mechanical properties of bulk-fill resin composites with single increment up to 4 mm: A novel mechanical strength test.

PURPOSE: To evaluate the mechanical properties of two brands of bulk-fill resin composites placed in a single increment up to 4 mm thickness via a novel mechanical strength test and provide related explanations. METHODS: Light transmission (LT), translucency parameter (TP), color difference (ΔE), Vickers hardness (HV) of two bulk-fill resin composites (Filtek Bulk Fill Posterior, Tetric N-Ceram Bulk Fill) and two conventional resin composites (Z100, Spectrum TPH) were evaluated. A novel flexural strength (FS) test method was applied for bulk-fill resin composite to determine the FS value of the bottom composites at depths of 1, 2, 3, and 4 mm after 24 hours of aging treatment (3 months water storage and 15,000 thermal cycles). The conventional resin composites were also tested for FS and all the FS results were subjected to Weibull analysis. Degree of conversion (DC) in the bulk-fill resin composites, light-cured at depths of 1, 2, 3, and 4 mm and conventional resin composites at depths of 2 and 4 mm, were assessed by FTIR. RESULTS: Both bulk-fill resin composites showed higher light transmission and translucency than that of conventional ones at each of the same thicknesses (1, 2, 3, 4 mm), wherein their flexural strength was not affected by depth. The Weibull analysis suggested both bulk-fill resin composites achieved good reliability and structural integrity under each curing thickness. Vickers hardness was affected by the material type and thickness. Bulk-fill resin composites showed a decrease in degree of conversion between 1 mm and 4 mm, but both were over 55%. CLINICAL SIGNIFICANCE: Filtek Bulk Fill Posterior, Tetric N-Ceram Bulk Fill achieved acceptable mechanical properties when cured at depths of up to 4 mm, which was beneficial from their optical and polymerized properties.

Heavy Metals in Soil around a Typical Antimony Mine Area of China: Pollution Characteristics, Land Cover Influence and Source Identification.

To understand contamination characteristics and identify sources of heavy metals in soil affected by complex mine activities, a detailed survey of soil heavy metals from different land cover types was investigated around the Xikuangshan (XKS) antimony mine in south-central China. Soil samples had average concentrations of Sb, As, Cd, Cr, Hg, Pb, Cu, Zn and Ni exceeding their background level in the Hunan province. Sb, As and Cd were the main pollutants. A total of 86.8% of samples were severely polluted, characterized by the Nemerow's comprehensive index, and 68.4% of samples were of very high potential ecological risk, primarily contributed by Sb, Cd and Hg. Among different land cover patterns, Hg, Pb and Cd concentrations showed a statistically significant difference. The application of Pearson correlation, principal component analysis (PCA) and hierarchical cluster analysis (HCA) combined with spatial interpolation GIS mapping revealed that Ni, Cr and Cu were mainly from natural parent materials, whereas other heavy metals were related to anthropogenic sources. Pb, As and Hg were mainly derived from smelting processes of sulfide minerals in the XKS area. The agricultural practice is the main factor for the accumulation of Cd and Zn, and sphalerite smelting also contributed to high Zn concentrations. Particularly, spatial variation of soil Sb concentrations was affected by multiple factors of complex antimony mine activities related to mining, beneficiation and smelting in the XKS area. These results are useful for the prevention and reduction of heavy metal contamination in soils by various effective measures in typical regions affected by antimony mine activities.

Pr3+-doped CeF3 crystals: Analysis of optical traits and fluorescence from Pr3+: 1D2 level for visible and near-infrared lasers.

In this work, four Pr3+ -doped CeF3 crystals with 0.2, 0.4, 0.6, and 0.8 at.% Pr3+ ion doping levels nominally in the melt have been successfully grown, and their absorption and emission spectra, including fluorescence decay times were analyzed. Especially, yellow and near-infrared (NIR) emissions originating from Pr3+: 1D2 level were explored in detail. In comparison, 0.2 at.% Pr3+-doped sample showed better spectral parameters in all studied crystals. In such a crystal, at 443 nm wavelength, the derived absorption cross-section (σabs) is 0.88 × 10-20 cm2 with full width at half maximum (FWHM) âˆ¼ 10 nm in π polarization direction, whereas σabs is 1.32 × 10-20 cm2 with FWHM âˆ¼ 6.7 nm in σ polarization direction at the same wavelength. Likewise, the calculated emission cross-section (σem) for 594 nm wavelength is 0.69 × 10-20 cm2 in π direction and 0.46 × 10-20 cm2 in σ direction. Also, acquired σem at 790, 755, 1014, and 1432 nm wavelengths is 6.15 × 10-21 cm2, 7.33 × 10-21 cm2, 7.66 × 10-21 cm2, and 6.56 × 10-21 cm2 individually. Here evaluated fluorescence decay time of 1D2 level is âˆ¼ 200.6 µs. Obtained higher σabs, larger σem, and higher luminescence decay rates of 0.2 at.% Pr: CeF3 crystal specify its potential as a gain medium for orange and NIR lasers.

Combined transcriptomic and metabolomic analysis of Salmonella in the presence or absence of PhoP-PhoQ system under low Mg2+ conditions.

INTRODUCTION: Previous reports revealed the role played by Salmonella PhoP-PhoQ system in virulence activation, antimicrobial tolerance and intracellular survival, the impact of PhoP-PhoQ on cell metabolism has been less extensively described. OBJECTIVES: The aim of this study is to address whether and how the PhoP-PhoQ system affects the cell metabolism of Salmonella. METHODS: We constructed a Salmonella phoP deletion mutant strain TT-81 (PhoP-OFF), a Salmonella PhoP constitutively expressed strain TT-82 (PhoP-ON) and a wild-type Salmonella PhoP strain TT-80 (PhoP-N), using P22-mediated generalized transduction or λ Red-mediated targeted mutagenesis. We then measured the in vitro growth kinetics of all test strains and determined their metabolomic and transcriptomic profiles using gas chromatography coupled with tandem mass spectrometry (GC-MS/MS) and RNA-seq technique, respectively. RESULTS: Low-Mg2+ conditions impaired the growth of the phoP deletion mutant strain TT-81 (PhoP-OFF) dramatically. 42 metabolites in the wild-type PhoP strain TT-80 (PhoP-N) and 28 metabolites in the PhoP constitutively expressed strain TT-82 (PhoP-ON) changed by the absence of phoP. In contrast, the level of 19 compounds in TT-80 (PhoP-N) changed comparing to the PhoP constitutively expressed strain TT-82 (PhoP-N). The mRNA level of 95 genes in TT-80 (PhoP-N) changed when phoP was disrupted, wherein 78 genes downregulated and 17 genes upregulated. 106 genes were determined to be differentially expressed between TT-81 (PhoP-OFF) and TT-82 (PhoP-ON). While only 16 genes were found to differentially expressed between TT-82 (PhoP-ON) and TT-80 (PhoP-N). CONCLUSION: Our findings confirmed the impact of PhoP-PhoQ system on lipopolysaccharide (LPS) modification, energy metabolism, and the biosynthesis or transport of amino acids. Most importantly, we demonstrated that the turnover of a given metabolite could respond differentially to the level of phoP. Taken together, the present study provided new insights into the adaptation of Salmonella to the host environment and helped to characterize the impact of the PhoP-PhoQ system on the cell metabolism.

Comparison of stress distribution between zirconia/alloy endocrown and CAD/CAM multi-piece zirconia post-crown: three-dimensional finite element analysis.

OBJECTIVES: This study aimed to evaluate a digital multi-piece zirconia post-crown to restore a mandibular second molar with extensive coronal loss and limited restoration space, and to compare the stress distribution between endocrowns made of zirconia or alloy and CAD/CAM multi-piece zirconia post-crowns. MATERIAL AND METHODS: Four three-dimensional finite element analysis models of a mandibular second molar with extensive coronal loss and limited restoration space were created as follows: (A) intact molar; (B) zirconia endocrown restored molar; (C) multi-piece post-crown restored-molar with tapered nail; (D) multi-piece post-crown restored molar with T-shaped nail. Models C and D were divided into two subgroups according to the material type: C1/D1, zirconia; C2/D2, NiCr alloy. The maximum modified von Mises failure criterion (mvM) stresses were calculated, and the stress distribution was recorded to analyze the effects of the restoration and material types on the biomechanical properties of dentin and prosthesis. RESULTS: The maximum mvM stress of dentin in model B (33.80 MPa) was lower compared with models C (C1, 37.81 MPa; C2, 36.36 MPa) and D (D1, 36.34 MPa; D2, 34.97 MPa) under vertical load, but the opposite was observed under oblique load. The highest mvM stress was concentrated in the nail region located in the root canal, and the T-shaped nail values were greater than the tapered nail, whereas model D with T-shaped nail showed a lower mvM stress level in dentin compared with Model C with tapered nail. CONCLUSIONS: The digital multi-piece zirconia post-crown is a potential approach to restore mandibular second molars with extensive coronal loss and limited restoration space. CLINICAL RELEVANCE: The digital multi-piece zirconia post-crown has potential to restore mandibular second molars with extensive coronal loss and limited restoration space using an innovative approach.

The DmeRF System Is Involved in Maintaining Cobalt Homeostasis in Vibrio parahaemolyticus.

Although cobalt (Co) is indispensable for life, it is toxic to cells when accumulated in excess. The DmeRF system is a well-characterized metal-response system that contributes to Co and nickel resistance in certain bacterial species. The Vibrio parahaemolyticus RIMD 2210633 genome also harbors a dmeRF operon that encodes a multiple antibiotic resistance regulator family transcriptional regulator and a cation diffusion facilitator family protein. Quantitative real-time PCR, growth curves analysis, inductively coupled plasma-mass spectrometry, ß-galactosidase activity assays, electrophoretic mobility shift assays, and a mouse infection experiment were performed to characterize the function of the DmeRF system in V. parahaemolyticus. Zinc, copper, and Co significantly increase dmeF expression, with Co inducing the greatest increase. DmeF promotes V. parahaemolyticus growth under high-Co conditions. Additionally, increased accumulation of cellular Co in the ΔdmeF mutant indicates that DmeF is potentially involved in Co efflux. Moreover, DmeR represses the dmeRF operon by binding directly to its promoter in the absence of Co. Finally, the DmeRF system was not required for V. parahaemolyticus virulence in mice. Collectively, our data indicate that the DmeRF system is involved in maintaining Co homeostasis in V. parahaemolyticus and DmeR functioning as a repressor of the operon.

Effect of the thicknesses of bulk-fill composites on bonding strength.

OBJECTIVES: This work evaluated the effects of thickness on resin bonding strength. METHODS: We set the two bulk-fill composites Filtek Bulk Fill Posterior (FBF) and Tetric N-Ceram Bulk Fill (TBF) as the experimental groups and the two conventional composites Filtek Z100 (Z100) and Spectrum TPH (ST) as the control groups. The translucency parameter (TP), color difference, Vickers hardness (HV), and microtensile bond strength (µTBS) of dentine and the resin composites were measured at different depths. RESULTS: In each group, TP and HV decreased with the increase in thickness. At the same depth, the TP of the bulk-fill composites was higher than that of the conventional composites. At the same depth, the HV of the four different resin composites followed the order of Z100>FBF>TBF>ST (except for the upper surface). Except for FBF at 3 and 4 mm, all of the other groups showed no visible color change at all the tested thicknesses. Although the µTBS values of the four different composites obtained through the bulk-fill technique were lower than those of composites obtained through the incremental fill technique, the µTBS of the bulk-fill composites obtained through the bulk-filling technique can reach 30 MPa. CONCLUSIONS: Bulk-fill composites applied in single 4 mm increments can meet the requirements of clinical application. However, the color stability of some bulk-fill composites cannot be maintained.

Evaluating the Bonding Performance of a Novel Dual-curing Composite Cement to Zirconia.

PURPOSE: To investigate the bond strength and durability of a novel dual-curing composite cement to zirconia under different curing conditions. MATERIALS AND METHODS: Zirconia plates of different thickness (0.5, 1, and 2 mm) were bonded with either a novel dual-curing composite cement (Panavia V5, PV5, Kuraray Noritake) or a traditional one (RelyX Ultimate, RUL, 3M Oral Care; Multilink Automix, MLA, Ivoclar Vivadent), in light-, self-, or dual-curing mode. Bonded specimens were subjected to shear bond strength (SBS) tests after 24 h of water storage or after artificially aging by 20,000 thermal cycles plus 150 days of water storage. The degree of conversion (DC) of the composite cements under different curing conditions was measured by Fourier transform infrared (FTIR) spectroscopy. The irradiance and translucency of the zirconia plates of different thickness were also investigated. RESULTS: The irradiance and translucency of zirconia decreased significantly with increasing thickness (p = 0.00). Both before and after aging, SBS of PV5 in self-curing mode was significantly higher than that of RUL (p = 0.07 before aging and 0.02 after aging) and MLA (p = 0.00 both before and after aging). However, for the three composite cements, light- and dual curing yielding the same SBSs for a constant Y-TZP thickness (p > 0.05). The FTIR analysis showed that, for all three dual-curing composite cements examined in this study, the mean DC values obtained in dual-curing mode were lower than those achieved in light-curing mode (p = 0.00 for PV5, RUL, and MLA). For RUL and MLA, lower mean DC values were obtained in self-curing than dual-curing mode (p = 0.00 for both RUL and MLA), while the DC values of PV5 showed no significant difference between self-curing and dual-curing mode (p = 0.33). CONCLUSION: When the photoactivation time is 60 s and the thickness of the zirconia restoration is less than 2 mm, it is safe to use the two traditional dual-curing composite cements RUL and MLA and PV5 for bonding zirconia. However, when the light exposure time is insufficient, PV5 provides improved bond strength and durability to zirconia.

Sex differences in the association between green tea consumption and hypertension in elderly Chinese adults.

BACKGROUND: Green tea has been one of the most popular beverages in China since ancient times. Mixed results concerning the effect of green tea consumption on the incidence of hypertension have been published over the past decades. However, no previous studies have focused on longevous individuals in China and the sex differences in the association between habitual green tea intake and hypertension. METHODS: The data extracted from the database of the Chinese Longitudinal Healthy Longevity Survey (CLHLS) in 2018 were used for a secondary analysis. Logistic regression models were employed to examine the odds ratio (OR) of daily green tea consumption on the incidence of hypertension by sex. RESULTS: A total of 9277 individuals were included in the analysis (39.8% were men). The included individuals had a mean age of 80.9 and 84.8 years for those who drank green tea daily and those who had never, respectively (p <  0.001). The incidence of hypertension varied at baseline according to green tea drinking habit and sex. For women who had a habitual green tea intake or had never drunk green tea, the incidence of hypertension was 47.3 and 43.9%, respectively (p = 0.241), whereas it was 51.6 and 39.7% for men (p <  0.001). After adjusting for potential confounders, a 38% increase in the risk of hypertension was observed in men who consumed green tea daily (OR, 1.38; 95% CI, 1.15-1.67; p <  0.001). CONCLUSIONS: Chinese longevous men had a 38% higher risk of developing hypertension when drinking green tea daily. However, no effect of green tea consumption on the incidence of hypertension in women was found. More attention should be paid to the lifestyle of longevous individuals for health promotion, and a sex-specific approach to deliver care for very elderly people is warranted.

Modifications of the Langendorff Method for Simultaneous Isolation of Atrial and Ventricular Myocytes from Adult Mice.

A single cardiomyocyte is a vital tool in the cellular and subcellular level studies of cardiac biology and diseases as a fundamental unit of contraction and electrical activity. Hence, isolating viable, high-quality cardiomyocytes from the heart is the initial and most crucial experimental step. Comparing the various protocols for isolating the cardiomyocytes of adult mice, the Langendorff retrograde perfusion is the most successful and reproducible method reported in the literature, especially for isolating ventricular myocytes. However, isolating quality atrial myocytes from the perfused heart remains challenging, and few successful isolation reports are available. Solving this complicated problem is extremely important because apart from ventricular disease, atrial disease accounts for a large part of heart diseases. Therefore, further investigations on the cellular level to reveal the mechanisms are warranted. In this paper, a protocol based on the Langendorff retrograde perfusion method is introduced and some modifications in the depth of aorta cannulation and the steps that may affect the digestion process to isolate atrial and ventricular myocytes were simultaneously made. Moreover, the isolated cardiomyocytes are confirmed to be amenable to patch clamp investigation.

Mechanical Properties of Nanohybrid Resin Composites Containing Various Mass Fractions of Modified Zirconia Particles.

PURPOSE: The aim of this study was to investigate the effect of various mass fractions of 10-methacry-loyloxydecyl dihydrogen phosphate (MDP)-conditioned or unconditioned zirconia nano- or micro-particles with different initiator systems on the mechanical properties of nanohybrid resin composites. METHODS: Both light-cured (L) and dual-cured (D) resin composites were prepared. When the mass fraction of the nano- or micro-zirconia fillers reached 55 wt%, resin composites were equipped with dual-cured initiator systems. We measured the three-point bending-strength, elastic modulus, Weibull modulus and translucency parameter of the nanohybrid resin composites containing various mass fractions of MDP-conditioned or unconditioned zirconia nano- or micro-particles (0%, 5 wt%, 10 wt%, 20 wt%, 30 wt% and 55 wt%). A Cell Counting Kit (CCK)-8 was used to test the cell cytotoxicity of the experimental resin composites. The zirconia nano- or micro-particles with MDP-conditioning or not were characterized by transmission electron microscopy (TEM), Fourier infrared spectroscopy (FTIR), and X-ray photoelectron spectroscopy (XPS). RESULTS: Resin composites containing 5-20 wt% MDP-conditioned or unconditioned nano-zirconia fillers exhibited better three-point bending-strength than the control group without zirconia fillers. Nano- or micro-zirconia fillers decreased the translucence of the nanohybrid resin composites. According to the cytotoxicity classification, all of the nano- or micro-zirconia fillers containing experimental resin composites were considered to have no significant cell cytotoxicity. The FTIR spectra of the conditioned nano- or micro-fillers showed new absorption bands at 1719 cm-1 and 1637 cm-1, indicating the successful combination of MDP and zirconia particles. The XPS analysis measured Zr-O-P peak area on MDP-conditioned nano- and micro-zirconia fillers at 39.91% and 34.89%, respectively. CONCLUSION: Nano-zirconia filler improved the mechanical properties of nanohybrid resin composites, but cannot be the main filler to replace silica filler. The experimental dual-cured composites can be resin cements with better opacity effects and a low viscosity.

Effect of APTES- or MPTS-Conditioned Nanozirconia Fillers on Mechanical Properties of Bis-GMA-Based Resin Composites.

To investigate the effects of 3-aminopropyltriethoxysilane (APTES)- or (3-mercaptopropyl)trimethoxysilane (MPTS)-conditioned nanozirconia fillers on the mechanical properties of Bis-GMA-based resin composites. The conditioned fillers were characterized by Fourier transform infrared (FTIR) spectroscopy, X-ray photoelectron spectroscopy (XPS), and thermodynamic calculations. They were then used to prepare Bis-GMA-based resin composites, whose flexural strength and elastic modulus were evaluated. The Cell Counting Kit-8 (CCK-8) assessed the composites' cytotoxicity. The FTIR spectra of the conditioned fillers showed new absorption bands at 1569 and 1100 cm-1, indicating successful grafting of APTES or MPTS onto nanozirconia. XPS confirmed the Zr-O-Si bonds in the APTES- or MPTS-conditioned fillers at contents of 2.02 and 6.98%, respectively. Thermodynamic calculations reaffirmed the chemical binding between the two silanes and nanozirconia fillers. Composites containing the conditioned nanozirconia fillers had significantly greater flexural strengths (APTES, 121.02 ± 8.31 MPa; MPTS, 132.80 ± 15.80 MPa; control, 94.84 ± 9.28 MPa) and elastic moduli (8.76 ± 0.52, 9.24 ± 0.60, and 7.44 ± 0.83 GPa, respectively) than a control with untreated fillers. The cytotoxicity assay identified no significant cytotoxicity by composites containing the conditioned fillers. Silanes were previously considered to be unable to chemically condition zirconia to bond with resin. Inclusion of APTES- or MPTS-conditioned nanozirconia fillers can improve the mechanical properties of Bis-GMA-based resin composites without obvious cytotoxicity in this study.

Evaluation of the interaction of chlorhexidine and MDP and its effects on the durability of dentin bonding.

OBJECTIVE: This study aimed to evaluate the potential interaction of chlorhexidine (CHX) and 10-methacryloyloxydecyl dihydrogen phosphate (MDP) and its effects on the durability of dentin bonding. METHODS: Two commercial adhesives were tested: a MDP-free adhesive (Single Bond 2, SB2) and a MDP-containing adhesive (Single Bond Universal, SBU). Teeth were randomly assigned to six groups and tested for micro-tensile bond strength (µTBS): Ctr, direct bonding with SB2; CHX, CHX conditioning and SB2; MDP, MDP conditioning and SB2; CHX+MDP, combined CHX and MDP conditioning and SB2; SBU, direct bonding with SBU; CHX+SBU, CHX conditioning and SBU. The potential interaction of CHX and MDP was assessed by measuring nanoleakage, in situ zymography, and chemoanalytic characterization via Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS) and nuclear magnetic resonance (NMR). Specimens for µTBS and nanoleakage tests were first subjected to water storage for 24h or 6 months. RESULTS: The initial µTBS values of the Ctr and CHX groups were significantly lower than those of the other four groups (P<0.05). Water storage for 6 months significantly weakened all groups (P<0.05), with the Ctr group showing the lowest µTBS. This group also showed more obvious nanoleakage than the other five groups. In situ zymography revealed that the Ctr group showed the strongest fluorescence and that the CHX+MDP group showed greater fluorescence than either CHX or MDP group. FTIR, XPS, and NMR indicated that MDP can interact with hydroxyapatite. NMR detected no Ca2+ salt peak for MDP when it was combined with CHX. SIGNIFICANCE: The application of either CHX or MDP alone can improve dentin bond durability. However, CHX may interfere with the formation of MDP-Ca salts.

Glycerol Phosphate Dimethacrylate: An Alternative Functional Phosphate Ester Monomer to 10-Methacryloyloxydecyl Dihydrogen Phosphate for Enamel Bonding.

The bonding performance of a glycerol phosphate dimethacrylate (GPDM)-based, two-step, self-etch (SE) adhesive was experimentally compared to that of 10-methacryloyloxydecyl dihydrogen phosphate (MDP)-based universal adhesives in different application modes for enamel bonding. Microtensile bond strength (µTBS) for adhesives bonded to enamel was measured initially (24 h water storage) and after 10 000 thermocycles plus water storage for 30 days. A GPDM-based, two-bottle, two-step, self-etch adhesive (Optibond Versa, OV) and three one-bottle MDP-based universal adhesives, one self-etching (Tetric N Bond Universal, TNBU) and two with etch-and-rinse (E&R) processing (Single Bond Universal (SBU); Clearfil Universal Bond Quick (CUBQ)), were tested. Scanning electron microscopy (SEM) evaluated nanoleakage at the bonding interfaces. A profilometer determined roughnesses of enamel surfaces after phosphoric acid etching, OV priming, or TNBU conditioning. SEM observed the corresponding surface morphology. NMR and X-ray photoelectron spectroscopy (XPS) characterized chemical bonding in hydroxyapatites (HAps) conditioned with the adhesives. Etch-and-rinse samples had significantly stronger bonding than self-etch samples (p < 0.05) irrespective of aging. The µTBS values for initial and aged OV were significantly higher than those of TNBU (p < 0.05). Aging did not significantly decrease µTBS for any sample except TNBU (p < 0.05), but it significantly aggravated nanoleakage. Etch-and-rinse processing resulted in less nanoleakage than self-etching; the OV samples leaked less than TNBU, both before and after aging. Phosphoric acid etching achieved the highest enamel surface roughness, followed by OV primer. Ca-O-P bonds in hydroxyapatite conditioned with TNBU, SBU, and CUBQ were confirmed by NMR, which showed similar results to XPS observations of conditioned hydroxyapatite powders except OV primer. The GPDM-based, two-step, self-etch adhesive can provide higher micromechanical retention potential, bond strength, and durability than the MDP-based universal adhesive in self-etch mode but lower performance than the MDP-based universal adhesive in etch-and-rinse mode. None of the tested adhesives could avoid nanoleakage after aging.

Effects of nano-zirconia fillers conditioned with phosphate ester monomers on the conversion and mechanical properties of Bis-GMA- and UDMA-based resin composites.

OBJECTIVES: This study investigated the effects of the surface conditioning of nano-zirconia fillers with 10-methacryloyloxydecyl dihydrogen phosphate (MDP) and dipentaerythritol penta-acrylate phosphate (PENTA) on the degree of conversion and mechanical properties of Bis-GMA- and UDMA-based resin composites containing the fillers. METHODS: MDP or PENTA conditioned nano-zirconia fillers were characterized by Fourier transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS). The conditioned nano-zirconia fillers were then used to prepare Bis-GMA- and UDMA-based resin composites. The bending strength and elastic modulus of the prepared composites were evaluated, and Weibull analysis was adopted to evaluate the reliability of the bending tests. Degree of conversion (DC) was measured by FTIR. RESULTS: FTIR and XPS analysis confirmed formation of ZrOP bonds in the MDP-conditioned and PENTA-conditioned zirconia fillers. MDP-conditioning and PENTA-conditioning increased bending strength of Bis-GMA-based composites (100.70 ±â€¯7.15 MPa, 98.65 ±â€¯4.76 Mpa) and UDMA-based composites (99.02 ±â€¯4.45 MPa, 99.92 ±â€¯5.78 MPa) compared to the groups with untreated nano-zirconia fillers (Bis-GMA-based, 84.98 ±â€¯5.13 MPa; UDMA-based, 88.24 ±â€¯4.61 MPa). The same trend was observed for the Weibull modulus (m) and elastic modulus. The MDP/PENTA conditioning did not affect the DC of the composites. CONCLUSION: MDP or PENTA conditioning of nano-zirconia fillers improved the mechanical properties of the composite compared to the composites containing untreated nano-zirconia fillers; however, it did not affect DC. CLINICAL SIGNIFICANCE: Mixing MDP- or PENTA-conditioned nano-zirconia fillers directly with gamma-MPS-conditioned silica to the resin composite has the potential to improve the composite's mechanical properties.