The possible role of sirtuin 5 in the pathogenesis of apical periodontitis.

OBJECTIVES: We investigated the relation between expression of sirtuin 5 (SIRT5) in osteoblastic cells and progression of apical periodontitis. The role of SIRT5 in hypoxia-induced reactive oxygen species (ROS) formation and osteoblast apoptosis was also examined. MATERIALS AND METHODS: Progression of rat apical periodontitis was monitored by conventional radiography and microcomputed tomography. SIRT5 and oxidative stress biomarker 8-OHdG in bone-lining cells were assessed by immunohistochemistry. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling was used to demonstrate apoptosis. In primary human osteoblasts cultured under hypoxia, Western blot was used to analyze SIRT5 expression and cleavage of pro-caspase 3 and poly(ADP-ribose) polymerase (PARP). SIRT5 was overexpressed through lentiviral technique. ROS formation and mitochondrial membrane potential changes were assessed by MitoSOX-Red and JC-1 fluorescence, respectively. Immunofluorescence microscope was used to evaluate mitochondrial release of cytochrome c. RESULTS: In rat apical periodontitis, disease progression was accompanied by decreased expression of SIRT5, increased oxidative stress, and enhanced apoptosis in bone-lining cells. SIRT5 was suppressed in cultured osteoblasts under hypoxia. SIRT5 overexpression ameliorated hypoxia-enhanced ROS formation, mitochondrial depolarization, cytochrome c leakage, activation of caspase-3, and PARP fragmentation. CONCLUSIONS: SIRT5 is able to alleviate hypoxia-enhanced osteoblast apoptosis. SIRT5 augmentation may have therapeutic potential for apical periodontitis.

ZnO Nanoparticles Induced Caspase-Dependent Apoptosis in Gingival Squamous Cell Carcinoma through Mitochondrial Dysfunction and p70S6K Signaling Pathway.

Zinc oxide nanoparticles (ZnO-NPs) are increasingly used in sunscreens, food additives, pigments, rubber manufacture, and electronic materials. Several studies have shown that ZnO-NPs inhibit cell growth and induce apoptosis by the production of oxidative stress in a variety of human cancer cells. However, the anti-cancer property and molecular mechanism of ZnO-NPs in human gingival squamous cell carcinoma (GSCC) are not fully understood. In this study, we found that ZnO-NPs induced growth inhibition of GSCC (Ca9-22 and OECM-1 cells), but no damage in human normal keratinocytes (HaCaT cells) and gingival fibroblasts (HGF-1 cells). ZnO-NPs caused apoptotic cell death of GSCC in a concentration-dependent manner by the quantitative assessment of oligonucleosomal DNA fragmentation. Flow cytometric analysis of cell cycle progression revealed that sub-G1 phase accumulation was dramatically induced by ZnO-NPs. In addition, ZnO-NPs increased the intracellular reactive oxygen species and specifically superoxide levels, and also decreased the mitochondrial membrane potential. ZnO-NPs further activated apoptotic cell death via the caspase cascades. Importantly, anti-oxidant and caspase inhibitor clearly prevented ZnO-NP-induced cell death, indicating the fact that superoxide-induced mitochondrial dysfunction is associated with the ZnO-NP-mediated caspase-dependent apoptosis in human GSCC. Moreover, ZnO-NPs significantly inhibited the phosphorylation of ribosomal protein S6 kinase (p70S6K kinase). In a corollary in vivo study, our results demonstrated that ZnO-NPs possessed an anti-cancer effect in a zebrafish xenograft model. Collectively, these results suggest that ZnO-NPs induce apoptosis through the mitochondrial oxidative damage and p70S6K signaling pathway in human GSCC. The present study may provide an experimental basis for ZnO-NPs to be considered as a promising novel anti­tumor agent for the treatment of gingival cancer.

Cyclic fatigue behavior of nickel-titanium dental rotary files in clinical simulated root canals.

BACKGROUND/PURPOSE: Dental rotary instruments can be applied in multiple conditions of canals, but unpredictable fatigue fracture may happen. This study evaluated the fatigue lives of two batches of nickel-titanium (NiTi) dental rotary files operating in clinically simulated root canals. METHODS: Single-step cyclic fatigue tests were carried out to assess the performance of two batches of NiTi files (ProTaper and ProFile) in nine combinations of simulated canals (cylinder radii 5 mm, 7.5 mm, and 10 mm, and insertion angles 20°, 40°, and 60°). Two-step cyclic fatigue tests were carried out in simulated root canals with the same radius by using the following two sets of insertion angles: (20°, 40°), (20°, 60°), (40°, 20°), and (60°, 20°). Fracture surfaces were observed by scanning electron microscopy. RESULTS: The single-step cyclic fatigue results showed that cyclic fatigue lives of the files decreased with increasing insertion angles or decreasing cylinder radius. The ProFile #25 .04 file was more fatigue resistant than the ProTaper F2 file. In two-step cyclic fatigue tests, the total fatigue lives were usually more than 100% when the files operated at a lower strain and then at a higher strain. By scanning electron microscopy, a larger area of fatigue striation corresponded to a longer fatigue life. CONCLUSION: Cyclic fatigue life can be influenced by the strains and geometries of files. The fatigue life was prolonged when the files operated at a lower strain and then at a higher strain. However, the fatigue life was shortened if the loading sequence was reversed.

Fracture resistance of dental nickel-titanium rotary instruments with novel surface treatment: Thin film metallic glass coating.

BACKGROUND/PURPOSE: Dental nickel-titanium (NiTi) rotary instruments are widely used in endodontic therapy because they are efficient with a higher success rate. However, an unpredictable fracture of instruments may happen due to the surface characteristics of imperfection (or irregularity). This study assessed whether a novel surface treatment could increase fatigue fracture resistance of dental NiTi rotary instruments. METHODS: A 200- or 500-nm thick Ti-zirconium-boron (Ti-Zr-B) thin film metallic glass was deposited on ProTaper Universal F2 files using a physical vapor deposition process. The characteristics of coating were analyzed by scanning electron microscopy, transmission electron microscopy, and X-ray diffractometry. In cyclic fatigue tests, the files were performed in a simulated root canal (radius=5 mm, angulation=60°) under a rotating speed of 300rpm. The fatigue fractured cross sections of the files were analyzed with their fractographic performances through scanning electron microscopy images. RESULTS: The amorphous structure of the Ti-Zr-B coating was confirmed by transmission electron microscopy and X-ray diffractometry. The surface of treated files presented smooth morphologies without grinding irregularity. For the 200- and 500-nm surface treatment groups, the coated files exhibited higher resistance of cyclic fatigue than untreated files. In fractographic analysis, treated files showed significantly larger crack-initiation zone; however, no significant differences in the areas of fatigue propagation and catastrophic fracture were found compared to untreated files. CONCLUSION: The novel surface treatment of Ti-Zr-B thin film metallic glass on dental NiTi rotary files can effectively improve the fatigue fracture resistance by offering a smooth coated surface with amorphous microstructure.

Erosive potential of soft drinks on human enamel: an in vitro study.

BACKGROUND/PURPOSE: Most soft drinks are acidic in nature. Regular consumption of these drinks may result in dental erosion. The aim of this in vitro study was to evaluate the erosive potential of different soft drinks in Taiwan by a novel multiple erosive method. METHODS: Four commercially available soft drinks in Taiwan were selected for this study. The properties of each product were analyzed to measure their pH, titratable acidity, and ion contents. The erosive potential of the soft drinks was measured based on the amount of loss of human enamel surface following its exposure to the soft drinks tested for different periods (20 minutes, 60 minutes, and 180 minutes). The enamel loss was measured using a confocal laser scanning microscope. RESULTS: The pH values of the soft drinks were below the critical pH value (5.5) for enamel demineralization, and ranged from 2.42 to 3.46. The drink with ingredients of citric acid and ascorbic acid had the highest titratable acidity (33.96 mmol OH(-)/L to pH 5.5 and 71.9 mmol OH(-)/L to pH 7). Exposure to all the soft drinks resulted in loss of human enamel surface (7.28-34.07 µm for 180-minute exposure). The beverage with the highest calcium content had the lowest erosive potential. CONCLUSION: All tested soft drinks were found to be erosive. Soft drinks with high calcium contents have significantly lower erosive potential. Low pH value and high citrate content may cause more surface enamel loss. As the erosive time increased, the titratable acidity to pH 7 may be a predictor of the erosive potential for acidic soft drinks. The erosive potential of the soft drinks may be predicted based on the types of acid content, pH value, titratable acidity, and ion concentration.

Prosthetic-Driven Autotransplantation with the Assistance of Medical Image-Processing Software and a Real-Time Navigation System: A Case Report.

Autotransplantation has been proven as a viable method of reconstructing missing teeth. While preparing the recipient site, the bone reduction location depends largely on the surgeon's experience. Inappropriate overpreparation can cause biologic and esthetic complications, such as buccal or lingual bone resorption. This paper provides an innovative method to aid clinicians in precisely preparing a recipient site with the assistance of medical image-processing software and a real-time navigation system. This case report presents the autotransplantation of a mandibular molar using this technique with good short-term (6 months) clinical outcomes, including radiographic bone fill, normal probing pocket depth, physiologic tooth mobility, acceptable gingival level, and satisfactory restoration.

Effects of ceramic thickness, ceramic translucency, and light transmission on light-cured bulk-fill resin composites as luting cement of lithium disilicate based-ceramics.

PURPOSE: To assess the effects of ceramic thickness, ceramic translucency, and light transmission on restorative composites used as luting cement for lithium disilicate-based ceramics. METHODS: Four luting types of cement were tested (n=8); a dual-cured resin cement (Multilink N), a light-cured conventional flowable composite (Tetric N-Flow), and two light-cured bulk-fill flowable composites (Tetric N-Flow Bulk Fill and X-tra base). The 20 s- or 40 s-light (1000 mW/cm2) was transmitted through 1- or 2-mm-thick high- or low-translucency (HT- or LT-) ceramic discs (IPS e.Max press) to reach the 1-mm-thick luting cement. Light transmitted to cement without ceramic served as a control. Vickers hardness number (VHN), flexural strength (FS), fractography, and degree of conversion (DC) were evaluated. One-way and multi-way analysis of variance was conducted to determine the effects of factors on VHN and FS. RESULTS: Ceramic thickness, light transmission time, and cement type significantly affected the VHN of the luting cement (P < .000). Only Multilink N (LT- and HT-1mm) and Tetric N-Flow (HT-1mm) reached 90% VHN of corresponding control by 20 s-light transmissions, but Tetric N-Flow exhibited lowest VHN and approximately 1/3-1/2 VHN of Multilink N (P < 0.05). X-tra base expressed superior physicochemical properties to Tetric N-Flow Bulk Fill (P < 0.05) and reached >90% VHN of control in all conditions with 40 s-light transmissions except for LT-2 mm. DC, FS, and fractography supported these findings. CONCLUSIONS: The light-cured bulk-fill composite served as a luting cement for lithium-disilicate-based ceramics in a product-dependent manner. Light transmission time is crucial to ensure sufficient luting cement polymerization.

Effects of surface treatments of bioactive tricalcium silicate-based restorative material on the bond strength to resin composite.

OBJECTIVES: This study aimed to enhance the bond strength between Biodentine™ (BD), a bioactive tricalcium silicate (C3S) based material, and resin composite through various surface treatments. METHODOLOGY: BD samples were immersed in either double distilled water or Hank's Balanced Salt Solution and analyzed using X-ray Diffraction (XRD). Shear bond strength (SBS) evaluations of BD were performed using Prime & Bond™ NT (PNT), Single Bond Universal (SBU), Xeno V (Xeno), and glass ionomer cement (GIC) following various etching durations (0 s/ 15 s/ 30 s/ 60 s with 37.5% phosphoric acid). Two primers, RelyX™ Ceramic Primer (RCP) and Monobond ™ Plus (MBP), were chosen to prime BD for SBS enhancement. Fractography and bonding interfaces were examined with energy dispersive X-ray spectroscopy (EDS)/ scanning electron microscopy (SEM) and Fourier-transform infrared spectroscopy (FTIR). RESULTS: XRD confirmed BD's main compositions as C3S, Ca(OH)2, CaCO3 and ZrO2 after 14 days crystal maturation. Etched BD did not improve SBS. GIC exhibited the lowest SBS (p < 0.05) among all adhesives, regardless of the etching mode (all < 1 MPa). The highest SBS (17.5 ± 3.6 MPa, p < 0.05) was achieved when BD primed with MBP followed by SBU application. FTIR and EDS showed γ-MPTS and10-MDP within the MBP primer interacted with C3S and ZrO2 of BD, achieving enhanced SBS. Most specimens exhibited mixed or cohesive failure modes. Significance BD's subpar mechanical properties and texture may contribute to its poor adhesion to resin composite. Pretreating BD with MBP primer, followed by SBU adhesive is recommended for improving bond strength.

[Effect of a 10-Methacryloyloxydecyl Dihydrogen Phosphate- treated Bioceramic Sealer on the Bond Strength of an Endodontic Fiber Post: Multilayer Composite Disk Models and Ultra-highspeed Imaging Analysis].

PURPOSE: Multiple materials are found in the root canal after fiber-post cementation. The layer of a bioceramic-based (BC) sealer may affect the bond strength (σBS) of the fiber post in the root canal. The purpose of this study was to employ multilayer compos-ite-disk models in diametral compression to investigate whether the bond strength between a fiber post and root dentin can be in-creased by the application of a primer on the BC sealer. MATERIALS AND METHODS: The multilayers of materials in the root canal required 3D finite-element (FE) stress analyses (FEA) to pro-vide precise σBS values. First, BC sealer was characterized using x-ray powder diffraction (XRD) to determine when the sealer com-pletely set and the types of crystals formed to select which primer to apply to the sealer. We selected a 10-methacryloyloxydecyl dihydrogen phosphate (10-MDP)-based primer to treat the BC sealer before post cementation. Ultra-highspeed (UHS) imaging was utilized to analyze the crack initiation interface. The obtained failure force was used in FE analysis to calculate σBS. RESULTS: UHS imaging validated the fracture interface at the post-dentin junction as FEA simulations predicted. σBS values of the fiber posts placed with various material combinations in the root canal were 21.1 ± 3.4 (only cement/ post), 22.2 ± 3.4 (BC sealer/cement/post) and 28.6 ± 4.3 MPa (10-MDP primer treated BC sealer/cement/post). The 10-MDP-treated BC sealer exhibited the highest σBS (p < 0.05). CONCLUSION: The multilayer composite disk model proved reliable with diametral compression testing. The presence of BC sealer in the root canal does not reduce σBS of the fiber post. Conditioning the BC sealer layer with 10-MDP primer before fiber-post cemen-tation increases σBS.

Multiple growth factors accommodated degradable submicron calcium sulfate hemihydrate/porous hydroxyapatite for dentin-pulp regeneration.

Vital pulp therapy (VPT) has gained significant consideration by utilizing the natural healing capacity of the inflamed pulp in healing process. However, the protective pulp capping materials that facilitate this healing process are still under investigation for the successful promotion of dentin-pulp regeneration. Herein, we developed a bioactive and biodegradable pulp capping material (denoted as sCSHA-GFs) by synthesizing inorganic submicron calcium sulfate hemihydrate (sCS)/porous hydroxyapatite (HA) loaded with growth factors (GFs) such as transforming growth factor-beta 1 (TGF-ß1), fibroblast growth factor-2 (FGF-2) and vascular endothelial growth factor (VEGF). Physiochemical characteristics of submicron CSHA-GFs (sCSHA-GFs) cement were determined. Human dental pulp stem cells (hDPSCs) were used for analyzing their biocompatibility and bioactivity for dentin mineralization. To evaluate the efficacy of sCSHA-GFs, we compared it with a commercial material, mineral trioxide aggregate (MTA), the reference standard used clinically on pulp capping. Our results showed that sCSHA-GFs cement presented good biodegradability with dissolution properties for sustained release of calcium (Ca2+) ions and GFs, and facilitated attachment, proliferation, differentiation and migration of hDPSCs. In addition, sCSHA-GFs cement was found to be more effective than MTA at prolonged incubation time in inducing the mRNA expression levels of odontoblastic differentiation markers, dentin sialophosphoprotein (DSPP) and dentin matrix protein (DMP-1), leading to increased mineralization (with calcium deposits) along with increased alkaline phosphatase (ALP) expressions, evident from Alizarin Red S and ALP staining assays. Our findings suggest that sCSHA-GFs cement may act as a suitable material in VPT for dentin-pulp regeneration.

External beam radiotherapy synergizes ¹88Re-liposome against human esophageal cancer xenograft and modulates ¹88Re-liposome pharmacokinetics.

External beam radiotherapy (EBRT) treats gross tumors and local microscopic diseases. Radionuclide therapy by radioisotopes can eradicate tumors systemically. Rhenium 188 ((188)Re)-liposome, a nanoparticle undergoing clinical trials, emits gamma rays for imaging validation and beta rays for therapy, with biodistribution profiles preferential to tumors. We designed a combinatory treatment and examined its effects on human esophageal cancer xenografts, a malignancy with potential treatment resistance and poor prognosis. Human esophageal cancer cell lines BE-3 (adenocarcinoma) and CE81T/VGH (squamous cell carcinoma) were implanted and compared. The radiochemical purity of (188)Re-liposome exceeded 95%. Molecular imaging by NanoSPECT/CT showed that BE-3, but not CE81T/VGH, xenografts could uptake the (188)Re-liposome. The combination of EBRT and (188)Re-liposome inhibited tumor regrowth greater than each treatment alone, as the tumor growth inhibition rate was 30% with EBRT, 25% with (188)Re-liposome, and 53% with the combination treatment at 21 days postinjection. Combinatory treatment had no additive adverse effects and significant biological toxicities on white blood cell counts, body weight, or liver and renal functions. EBRT significantly enhanced the excretion of (188)Re-liposome into feces and urine. In conclusion, the combination of EBRT with (188)Re-liposome might be a potential treatment modality for esophageal cancer.