Autophagy Alters the Susceptibility of Candida albicans Biofilms to Antifungal Agents.

Candida albicans (C. albicans) reigns as a major cause of clinical candidiasis. C. albicans biofilms are known to increase resistance to antifungal agents, making biofilm-related infections particularly challenging to treat. Drug resistance is of particular concern due to the spread of multidrug-resistant fungal pathogens, while autophagy is crucial for the maintenance of cellular homeostasis. Therefore, this study aimed to investigate the effects of an activator and an inhibitor of autophagy on the susceptibility of C. albicans biofilms to antifungal agents and the related mechanisms. The susceptibility of C. albicans biofilms to different antifungal agents after treatment with or without the autophagy activator or inhibitor was evaluated using XTT assay. Alkaline phosphatase (ALP) activity and reactive oxygen species (ROS) level, as well as the expression of ROS-related and autophagy-related genes, were examined to evaluate the autophagic activity of C. albicans biofilms when treated with antifungal agents. The autophagosomes were observed by transmission electron microscopy (TEM). The susceptibility of C. albicans biofilms to antifungal agents changed when autophagy changed. The ALP activity and ROS level of C. albicans biofilms increased with the treatment of antifungal agents, and autophagosomes could be observed in C. albicans biofilms. Autophagy was involved in the susceptibility of C. albicans biofilms to antifungal agents.

Apically extruded debris, canal transportation, and shaping ability of nickel-titanium instruments on contracted endodontic cavities in molar teeth.

PURPOSE: Apically extruded debris, canal transportation and shaping ability were compared between contracted endodontic cavities (CECs) and traditional endodontic cavities (TECs) after instrumentation with XP-endo Shaper (XPS), ProTaper Gold (PTG), ProTaper for hand-use (HPT) and Hero Shaper. METHODS: The CECs or TECs groups were sub-divided into 24 groups according to root canal morphology and nickel-titanium (Ni-Ti) instruments. The weight of apically extruded debris was calculated using the Myers and Montgomery model. Pre- and postoperative images of teeth were scanned using micro-CT and the three-dimensional models were constructed and compared. RESULTS: Under CECs or TECs, XPS and PTG produced less apical debris and formed less canal transportation than HPT and Hero Shaper (P < 0.05). XPS group under CECs extruded less apical debris than that under TCEs for round canals with curvature of 20°-35° (P < 0.05). The centering ratios of four tested instruments were higher under TECs than those under CECs (P < 0.05). The HPT and Hero Shaper had more transportation under CECs than that under TCEs (P < 0.05). No statistical difference was found regarding shaping ability among all the groups. CONCLUSION: Under CECs, XPS preserves the original root canal anatomy, meanwhile it produces less apical debris than the other instruments.

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.

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.

Effects of hydrothermal aging, thermal cycling, and water storage on the mechanical properties of a machinable resin-based composite containing nano-zirconia fillers.

OBJECTIVES: This study aimed to investigate the effects of hydrothermal aging, thermal cycling, and water storage on the mechanical properties of a machinable resin-based composite containing nano-zirconia fillers. MATERIALS AND METHODS: A machinable resin-based composite containing nano-zirconia fillers (Lava Ultimate, LU) and a resin-based composite with a similar resin matrix-to-filler ratio but without zirconia fillers (Tetric N-Ceram, TNC) were prepared into bars and assigned into four groups based on the type of aging treatment (hydrothermal aging, thermal cycling, water storage, or no aging). The phase transformations of the zirconia fillers in LU after aging were evaluated by X-ray diffraction. The flexural strength, Weibull modulus, flexural modulus, and Vickers hardness of each group were investigated. The fracture surface morphologies of both resin-based composites before and after aging were observed by a scanning electron microscopy (SEM). RESULTS: Only Tetragonal zirconia was detected in the LU samples. Both before and after aging, the flexural strength, flexural modulus, and Vickers hardness values of LU were significantly higher than those of TNC (p < 0.05) with the exception of the flexural modulus of LU, which showed no difference with that of TNC after water storage (p = 0.68). Hydrothermal aging, thermal cycling, and water storage had no significant effects on the surface Vickers hardnesses of LU or TNC (p > 0.05). Hydrothermal aging significantly improved the flexural strength of LU (p = 0.00). Thermal cycling (p = 0.00) and water storage (p = 0.00) significantly decreased the flexural strength of LU. The flexural strength of TNC was not decreased by hydrothermal aging (p = 0.82) or water storage (p = 0.36), while it was decreased by thermal cycling (p = 0.00). The hydrothermal aging group of LU exhibited the highest Weibull modulus. CONCLUSIONS: The machinable resin-based composite containing nano-zirconia fillers provides superior flexural strength, flexural modulus, and Vickers hardness compared to the direct-filling resin-based composite with a similar resin matrix-to-filler ratio, although it fails to provide better aging resistance.