siRNA high throughput screening identifies regulators of chloropicrin and hydrogen fluoride injury in human corneal epithelial cell models.

Corneal injuries induced by various toxicants result in similar clinical presentations such as corneal opacity and neovascularization. Many studies suggest that several weeks post-exposure a convergence of the molecular mechanisms drives these progressive pathologies. However, chemical agents vary in toxicological properties, and early molecular responses are anticipated to be somewhat dissimilar for different toxicants. We chose 3120 targets from the Dharmacon Human Druggable genome to screen for chloropicrin (CP) and hydrogen fluoride (HF) corneal injury as we hypothesized that targets identified in vitro may be effective as therapeutic targets in future studies. Human immortalized corneal epithelial cells (SV40-HCEC) were used for screening. Cell viability and IL-8 were analyzed to down-select hits into validation studies, where multiplex cytokine analysis and high content analysis were performed to understand toxicant effect and target function. Some endpoints were also evaluated in a second human immortalized corneal epithelial cell line, TCEpi. Over 20 targets entered validation studies for CP and HF; of these, only three targets were shared: NR3C1, RELA, and KMT5A. These findings suggest that early molecular responses to different toxicants may be somewhat distinctive and present dissimilar targets for possible early intervention.

Biocompatibility Analyses of HF-Passivated Magnesium Screws for Guided Bone Regeneration (GBR).

Background: Magnesium (Mg) is one of the most promising materials for human use in surgery due to material characteristics such as its elastic modulus as well as its resorbable and regenerative properties. In this study, HF-coated and uncoated novel bioresorbable magnesium fixation screws for maxillofacial and dental surgical applications were investigated in vitro and in vivo to evaluate the biocompatibility of the HF coating. Methods: Mg alloy screws that had either undergone a surface treatment with hydrofluoric-acid (HF) or left untreated were investigated. In vitro investigation included XTT, BrdU and LDH in accordance with the DIN ISO 10993-5/-12. In vivo, the screws were implanted into the tibia of rabbits. After 3 and 6 weeks, degradation, local tissue reactions and bony integration were analyzed histopathologically and histomorphometrically. Additionally, SEM/EDX analysis and synchrotron phase-contrast microtomography (µCT) measurements were conducted. The in vitro analyses revealed that the Mg screws are cytocompatible, with improved results when the surface had been passivated with HF. In vivo, the HF-treated Mg screws implanted showed a reduction in gas formation, slower biodegradation and a better bony integration in comparison to the untreated Mg screws. Histopathologically, the HF-passivated screws induced a layer of macrophages as part of its biodegradation process, whereas the untreated screws caused a slight fibrous tissue reaction. SEM/EDX analysis showed that both screws formed a similar layer of calcium phosphates on their surfaces and were surrounded by bone. Furthermore, the µCT revealed the presence of a metallic core of the screws, a faster absorbing corrosion front and a slow absorbing region of corroded magnesium. Conclusions: Overall, the HF-passivated Mg fixation screws showed significantly better biocompatibility in vitro and in vivo compared to the untreated screws.

GPI7-mediated glycosylphosphatidylinositol anchoring regulates appressorial penetration and immune evasion during infection of Magnaporthe oryzae.

Glycosylphosphatidylinositol (GPI) anchoring plays key roles in many biological processes by targeting proteins to the cell wall; however, its roles are largely unknown in plant pathogenic fungi. Here, we reveal the roles of the GPI anchoring in Magnaporthe oryzae during plant infection. The GPI-anchored proteins were found to highly accumulate in appressoria and invasive hyphae. Disruption of GPI7, a GPI anchor-pathway gene, led to a significant reduction in virulence. The Δgpi7 mutant showed significant defects in penetration and invasive growth. This mutant also displayed defects of the cell wall architecture, suggesting GPI7 is required for cell wall biogenesis. Removal of GPI-anchored proteins in the wild-type strain by hydrofluoric acid (HF) pyridine treatment exposed both the chitin and ß-1,3-glucans to the host immune system. Exposure of the chitin and ß-1,3-glucans was also observed in the Δgpi7 mutant, indicating GPI-anchored proteins are required for immune evasion. The GPI anchoring can regulate subcellular localization of the Gel proteins in the cell wall for appressorial penetration and abundance of which for invasive growth. Our results indicate the GPI anchoring facilitates the penetration of M. oryzae into host cells by affecting the cell wall integrity and the evasion of host immune recognition.

Strategic design of a Mussel-inspired in situ reduced Ag/Au-Nanoparticle Coated Magnesium Alloy for enhanced viability, antibacterial property and decelerated corrosion rates for degradable implant Applications.

Magnesium (Mg) and its alloys have attracted much attention as a promising candidate for degradable implant applications however the rapid corrosion of magnesium inside the human body greatly limits its use as an implant material. Therefore, coating the alloy surface with a multifunctional film is a promising way to overcome the drawbacks. Here we propose for the first time a multifunction layer coating to enhance the cell viability, antibacterial property and decelerated corrosion rates to act as a novel material to be used for degradable implant Applications. For that, the magnesium alloy (AZ31) was first treated with hydrofluoric acid (HF) and then dopamine tris Hydrochloric acid (tris-HCL) solution. The reducing catechol groups in the polydopamine (PD) layer subsequently immobilize silver/gold ions in situ to form uniformly dispersed Ag/Au nanoparticles on the coating layer. The successful formation of Ag/Au nanoparticles on the HF-PD AZ31 alloy was confirmed using XPS and XRD, and the morphology of all the coated samples were investigated using SEM images. The alloy with HF-PDA exhibit enhanced cell attachment and proliferation. Moreover, the nanoparticle immobilized HF-PD alloy exhibited dramatic corrosion resistance enhancement with superior antibacterial properties and accountable biocompatibility. Thus the result suggest that HF-PD Ag/Au alloy has great potential in the application of degradable implant and the surface modification method is of great significance to determine its properties.

The influence of chemical polishing of titanium scaffolds on their mechanical strength and in-vitro cell response.

Selective Laser Melting (SLM) is a powder-bed-based additive manufacturing method, using a laser beam, which can be used to produce metallic scaffolds for bone regeneration. However, this process also has a few disadvantages. One of its drawbacks is the necessity of post-processing in order to improve the surface finish. Another drawback lies in the removal of unmelted powder particles from the build. In this study, the influence of chemical polishing of SLM fabricated titanium scaffolds on their mechanical strength and in vitro cellular response was investigated. Scaffolds with bimodal pore size (200 µm core and 500 µm shell) were fabricated by SLM from commercially pure titanium powder and then chemically treated in HF/HNO3 solutions to remove unmelted powder particles. The cell viability and mechanical strength were compared between as-made and chemically-treated scaffolds. The chemical treatment was successful in the removal of unmelted powder particles from the titanium scaffold. The Young's modulus of the fabricated cellular structures was of 42.7 and 13.3 GPa for as-made and chemically-treated scaffolds respectively. These values are very similar to the Young's modulus of living human bone. Chemical treatment did not affect negatively cell proliferation and differentiation. Additionally, the chemically-treated scaffolds had a twofold increase in colonization of osteoblast cells migrating out of multicellular spheroids. Furthermore, X-ray computed microtomography confirmed that chemically-treated scaffolds met the dimensions originally set in the CAD models. Therefore, chemical-treatment can be used as a tool to cancel the discrepancies between the designed and fabricated objects, thus enabling fabrication of finer structures with regular struts and high resolution.

Comparative study of the effect of Er:YAG and Er:Cr;YSGG lasers on porcelain: etching for the bonding of orthodontic brackets.

This study investigated the effect of Er:YAG (smart 2940 Dplus, DEKA, Italy) and Er:CrYSGG (Waterlase iPlus, Biolase, USA) lasers on the shear bond strength (SBS) between the orthodontic brackets and dental porcelain in comparison with conventional acid etching with 9% hydrofluoric acid (HF, Ultradent, USA). A total of 60 specimens of maxillary incisor crown were prepared and randomly assigned to five groups; each group was subjected to a different porcelain surface conditioning: (1) etching with the 9% HF for 2 min; (2) etching with the 9% HF for 2 min followed by irradiation with the Er:CrYSGG laser (3-W power, 10-Hz frequency for 10 s); (3) etching with the 9% HF for 2 min followed by irradiation with the Er:YAG laser (3-W power, 10-Hz frequency for 10 s); (4) Irradiation with the Er:CrYSGG laser (3-W power, 10-Hz frequency for 10 s without acid etching) and (5) irradiation with the Er:YAG laser (3-W power,10-Hz frequency for 10 s without acid etching). After using Transbond XT primer and Transbond XT adhesive, the metal brackets (Dentaurum, Germany equilibrium 2, optimal design) bonded to the conditioned porcelain surface. Subsequently, the specimens were thermocycled for 5000 cycles and then debonded using the Universal Testing Machine (Zwick). In each group, one specimen was not bonded to brackets to allow further examination with electron microscopy. After debonding, the specimens were examined by stereomicroscope to determine their adhesive remnant index (ARI). The average SBS [Mean (SD)] values in the five groups were as follows: HF (32.58 ± 9.21 MPa), Er:CrYSGG + HF (27.81 ± 7.66 MPa), Er:YAG + HF (23.08 ± 9.55 MPa), Er:CrYSGG (14.11 ± 9.35 MPa), and Er:YAG (6.30 ± 3.09 MPa). A statistically significant difference in SBS existed between the first three groups and the two laser groups (df = 4, F = 18.555, p < 0.001). Evaluation of ARI values showed that bond failures in the first three groups were mostly of cohesive and mixed types, but in the laser groups, they were mostly adhesive. Chi-square was not significant between groups (p = 0.219). The Er:YAG laser with the stated specifications is not a suitable alternative to HF etching. In the case of Er:CrYSGG laser, although the conditioning outcome met the bond strength requirement for orthodontic brackets (that is, 6-8 MPa). Therefore, the bond strength must be further improved by fine-tuning the irradiation details.

Osteogenic activity of titanium surfaces with hierarchical micro-/nano-structures obtained by hydrofluoric acid treatment.

An easier method for constructing the hierarchical micro-/nano-structures on the surface of dental implants in the clinic is needed. In this study, three different titanium surfaces with microscale grooves (width 0.5-1, 1-1.5, and 1.5-2 µm) and nanoscale nanoparticles (diameter 20-30, 30-50, and 50-100 nm, respectively) were obtained by treatment with different concentrations of hydrofluoric acid (HF) and at different etching times (1%, 3 min; 0.5%, 12 min; and 1.5%, 12 min, respectively; denoted as groups HF1, HF2, and HF3). The biological response to the three different titanium surfaces was evaluated by in vitro human bone marrow-derived mesenchymal stem cell (hBMMSC) experiments and in vivo animal experiments. The results showed that cell adhesion, proliferation, alkaline phosphatase activity, and mineralization of hBMMSCs were increased in the HF3 group. After the different surface implants were inserted into the distal femurs of 40 rats, the bone-implant contact in groups HF1, HF2, and HF3 was 33.17%±2.2%, 33.82%±3.42%, and 41.04%±3.08%, respectively. Moreover, the maximal pullout force in groups HF1, HF2, and HF3 was 57.92±2.88, 57.83±4.09, and 67.44±6.14 N, respectively. The results showed that group HF3 with large micron grooves (1.5-2.0 µm) and large nanoparticles (50-100 nm) showed the best bio-functionality for the hBMMSC response and osseointegration in animal experiments compared with other groups.

Effect of Nanofeatured Topography on Ceria-Stabilized Zirconia/Alumina Nanocomposite on Osteogenesis and Osseointegration.

PURPOSE: The objective of this study was to evaluate the osteogenic and osseointegration capability of the Ce-tetragonal zirconia polycrystal (TZP)-based nanostructured zirconia/alumina composite (Ce-TZP/Al2O3) that was treated with hydrofluoric acid (HF). MATERIALS AND METHODS: Osteogenic MC3T3-E1 cells were cultured on acid-etched titanium (AETi) disks and Ce-TZP/Al2O3 disks without HF treatment (Zr[0%]), with 4% HF treatment (Zr[4%]), or with 55% HF treatment (Zr[55%]) for 24 hours, and biologic responses were compared among four conditions in vitro. Miniature implants of AETi and Zr(55%) were surgically placed in the femora of rats. Osseointegration was evaluated by a biomechanical push-in test after 2 and 4 weeks of healing. RESULTS: The surface of Zr(55%) rendered nanofeatured topography with a greater surface area and roughness, and extensive geographical undercut as ceria-zirconia crystal disappeared from the superficial layer and was similar to the surface morphology of biomineralized matrices. Culture studies showed that the attachment, proliferation, spread, and functional phenotypes of osteogenic cells, such as alkaline phosphatase activity and bone-related gene expression, were remarkably increased on the Zr(55%) surface. The strength of osseointegration measured using the biomechanical push-in test in a rat model was stronger for Zr(55%) implants than for AETi implants by 1.6 fold. CONCLUSION: The nanostructured Ce-TZP/ Al2O3 surface substantially enhanced the osteogenic response in vitro and the osseointegration capability in vivo, which suggest its potential clinical application as a novel implant material.

[Effect of hydrofluoric acid concentration on the surface morphology and bonding effectiveness of lithium disilicate glass ceramics to resin composites].

OBJECTIVE: This study aimed at determining the influence of hydrofluoric acid (HF) in varied concentrations on the surface morphology of lithium disilicate glass ceramics and bond durability between resin composites and post-treated lithium disilicate glass ceramics. METHODS: After being sintered, ground, and washed, 72 as-prepared specimens of lithium disilicate glass ceramics with dimensions of 11 mm×13 mm×2 mm were randomly divided into three groups. Each group was treated with acid solution [32% phosphoric acid (PA) or 4% or 9.5% HF] for 20 s. Then, four acidified specimens from each group were randomly selected. One of the specimens was used to observe the surface morphology using scanning electron microscopy, and the others were used to observe the surface roughness using a surface roughness meter (including Ra, Rz, and Rmax). After treatment with different acid solutions in each group, 20 samples were further treated with silane coupling agent/resin adhesive/resin cement (Monobond S/Multilink Primer A&B/Multilink N), followed by bonding to a composite resin column (Filtek™ Z350) with a diameter of 3 mm. A total of 20 specimens in each group were randomly divided into two subgroups, which were used for measuring the microshear bond strength, with one of them subjected to cool-thermal cycle for 20 000 times. RESULTS: The surface roughness (Ra, Rz, and Rmax) of lithium disilicate glass ceramics treated with 4% or 9.5% HF was significantly higher than that of the ceramic treated with PA (P<0.05). The lithium disilicate glass ceramics treated with 9.5% HF also demonstrated better surface roughness (Rz and Rmax) than that of the ceramics treated with 4% HF. Cool-thermal cycle treatment reduced the bond strength of lithium disilicate glass ceramics in all groups (P<0.05). After cool-thermal cycle, the lithium disilicate glass ceramics treated with HF had higher bond strength than that of the ceramics treated with PA. The lithium disilicate glass ceramics treated with 4% HF had higher bond strength than that of the ceramics treated with 9.5% HF (P<0.05). During cool-thermal cycle, the lithium disilicate glass ceramics treated with 4% HF demonstrated higher reduction in bond strength than that of the samples treated with 9.5% HF (P<0.05). CONCLUSIONS: The concentration of HF significantly affected the surface morphology of lithium disilicate glass ceramics and the bond durability between resin composites and post-treated lithium disilicate glass ceramics. The bond strength between resin composites and post-treated lithium disilicate glass ceramic was more efficiently maintained by treatment with 9.5% HF.

Influence of post-etching surface treatment and thermo-mechanical cycling on fracture strength of ceramics.

BACKGROUNG: The aim of this study was to evaluate fracture strength of lithium disilicate-based ceramic crowns submitted to neutralization of hydrofluoric acid (HF) precipitates associated with ultrasonic bath and to thermo-mechanical cycling. METHODS: Eighty human molars received conventional full crown preparation after being included in polyurethane standard to simulate periodontal ligament. After scanning, the ceramic blocks were machined using CAD/CAM system to obtain the crowns. The crowns were distributed according to surface treatment: S and S-C; HF and silane; SNU and SNU-C; HF neutralization of HF precipitates, ultrasonic bath and silane. The crowns were cemented with dual cure resin cement and the specimens in Groups S-C and SNU-C were submitted to thermo-mechanical cycling (5/55 °C/60 s while 1.2 million mechanical cycles of 4 Hz/100 N). The samples were submitted to compressive test. Analysis of fractures was performed macroscopically (Burke method) and using Scanning Electron Microscope. The data of fracture strength were analyzed using the two-way ANOVA. RESULTS: No statistical difference among the groups was found (p-value = 0,799). Most failures were associated with the remaining tooth fracture and the fracture origin was located on the cementation surface. CONCLUSIONS: Postetching cleaning protocols do not improve the fracture strength of ceramics crowns. Thermo-mechanical aging did not weakened the ceramic crown.

Identification of glycosylphosphatidylinositol-anchored proteins and ω-sites using TiO2-based affinity purification followed by hydrogen fluoride treatment.

Glycosylphosphatidylinositol anchored proteins (GPI-APs) in the outer leaflet of the membrane microdomains, commonly referred to as lipid rafts, play important roles in many biological processes such as signal transduction, cell adhesion, protein trafficking, and antigen presentation. From a topological viewpoint, elucidating the presence and localization of GPI-anchor modification sites (ω-sites) is important for the study of the biophysical properties and anchoring mechanisms of these proteins. However, very few reports have actually identified ω-sites of GPI-APs. To enable large-scale site-specific analysis of GPI anchoring, we developed a method for identification of ω-sites by mass spectrometry by combining titanium dioxide-based affinity purification and hydrogen fluoride treatment. This method was able to identify ~3-fold more GPI-APs than our previous method: the new technique identified a total of 73 ω-sites derived from 49 GPI-APs. In 13 of the 49 GPI-APs identified, the GPI-anchor attached to multiple amino acids in the C-terminal site, yielding a variety of different protein species. This method allows us to simultaneously identify many GPI-AP protein species with different ω-sites. We also demonstrated the C-terminal GPI anchor attachment signal peptide, based on information about the GPI anchor binding sites of 49 GPI-APs. Thus, our results provide evidence for new insight into the GPI-anchored proteome and the role of GPI anchoring. BIOLOGICAL SIGNIFICANCE: GPI-anchored proteins (GPI-APs) are localized to the outer leaflet of the plasma membranes. Because the GPI anchor is a complex structure, the identification of GPI-anchored peptides by mass spectrometry has always been considered difficult. To improve the feasibility of large-scale site-specific analysis of GPI anchoring, we developed a method for identification of GPI-anchored peptides by combining titanium dioxide-based affinity purification with hydrogen fluoride treatment. Using this novel technique, we identified a total of 73 ω-sites derived from 49 GPI-APs. These data may help us to develop a comprehensive understanding of the GPI-anchored proteome and the role of GPI anchoring. Moreover, this method could be used to discover GPI-APs as candidate biomarkers.

Effects of Acid Treatment on Dental Zirconia: An In Vitro Study.

The aim of this study was to evaluate the effects of hydrofluoric (HF) acid, acetic acid, and citric acid treatments on the physical properties and structure of yttria-stabilized tetragonal zirconia polycrystal (Y-TZP) at ambient temperature. In total, 110 bar-shaped zirconia specimens were randomly assigned to 11 groups. The specimens in the control group (C) received no surface treatment, while those in the Cage group were hydrothermally aged at 134°C and 0.2 MPa for 20 h. Ten specimens each were immersed at ambient temperature in 5% and 40% HF acid for 2 h (40HF0), 1 day (5HF1, 40HF1), and 5 days (5HF5, 40HF5), while 10 each were immersed at ambient temperature in 10% acetic acid and 20% citric acid for 7 (AC7, CI7) and 14 days (AC14, CI14). X-ray diffraction (XRD) was used to quantitatively estimate the monoclinic phase. Furthermore, flexural strength, surface roughness, and surface Vickers hardness were measured after treatment. Scanning electron microscopy (SEM) was used to characterize the surface morphology. The Cage group specimens exhibited an increased monoclinic phase and flexural strength. Furthermore, 40% HF acid immersion decreased the flexural strength and surface hardness and deteriorated the surface finish, while 5% HF acid immersion only decreased the surface hardness. All the HF acid-immersed specimens showed an etched surface texture on SEM observations, while the other groups did not. These findings suggest that the treatment of Y-TZP with 40% HF acid at ambient temperature causes potential damage, while treatment with 5% HF acid, acetic acid, and citric acid is safe.

Effect of different cleaning regimens on the adhesion of resin to saliva-contaminated ceramics.

PURPOSE: The aim of this study was to evaluate the influence of different cleaning regimens on the microshear bond strength (µSBS) of three different all-ceramic surfaces after saliva contamination. MATERIAL AND METHODS: Cubic ceramic specimens (3 × 3 × 3 mm(3) ) were prepared from three types of ceramics: zirconium dioxide (Z), leucite-reinforced glass ceramic (E), lithium disilicate glass ceramic (EX; n = 12/subgroup). A total of 144 composite resin cylinders (diameter: 1 mm, height: 3 mm) were prepared. Three human-saliva-contaminated surfaces of ceramic specimens were cleaned with either water spray (WS), with 0.5% sodium hypochlorite solution (HC), or with a cleaning paste (CP). Control surface (C) was not contaminated or cleaned. Composite cylinders were bonded to each surface with a resin luting cement. All specimens were stored at 37°C in deionized water until fracture testing. µSBS tests were performed in a universal testing machine (0.5 mm/min), and the results (MPa ± SD) were statistically analyzed (two-way ANOVA, Bonferroni a = 0.05). Fractured surfaces were analyzed to identify the failure types using an optical microscope at 50× magnification. Two representative specimens from all groups were examined with scanning electron microscopy. RESULTS: µSBS test results were significantly affected by the saliva cleaning regimens (p = 0.01) and the ceramic types (p = 0.03). The interaction terms between the ceramic type and saliva cleaning regimen were also significant (p < 0.05). There were no significant differences among the µSBS values (MPa ± SD) for the Z group (C = 17.5 ± 8.8; WS = 16.0 ± 4.9; HC = 17.6 ± 5.8; CP = 16.6 ± 7.5; p > 0.05). In the EX group, C resulted in significantly higher µSBS values (32.6 ± 7.4) than CP (17.4 ± 8.9), WS (15.6 ± 7.3), and HC (14.3 ± 4.5) (p < 0.05); however, C (20.4 ± 7.1) and HC (19.2 ± 7.5) showed higher µSBS values than CP (13.8 ± 4.8) and WS (10.9 ± 5.7) in the E group. Some cohesive failures within the luting resin were observed in the E and EX groups, whereas only adhesive failures were seen in zirconia groups for all surface treatments. CONCLUSIONS: Different ceramic surface cleaning regimens after saliva contamination of the zirconium dioxide revealed µSBS similar to the control group, whereas all surface cleaning regimens tested significantly decreased the bond strength values in the lithium disilicate glass ceramic. The leucite-reinforced glass-ceramic group benefited from 0.5% sodium hypochlorite solution cleaning with increased bond strengths. CLINICAL SIGNIFICANCE: Adhesive cementation of zirconia presents a clinically challenging protocol, and the cementation surface contamination of the zirconia restorations and the inadequate removal of the contaminants increase the risk of failure, as for all ceramic types. This study demonstrated that surface cleaning regimens should be applied according to different ceramic properties.

[Effect of SiO2-ZrO2slurry coating on surface performance of zirconia ceramic].

OBJECTIVE: To evaluate the effect of SiO2-ZrO2slurry coating on surface performance of zirconia ceramic. METHODS: Seventy pre-sintered zirconia discs were randomly divided into seven groups with 10 discs per group. Sample discs in each group received one of the following seven different surface treatments, namely, sintered (group AS), sand blasting after sintered (group SB), coated with slurry of mole ratio of SiO2to ZrO22:1 (group 2SiO2-1ZrO2), coated with slurry of mole ratio of SiO2to ZrO21:1 (group 1SiO2-1ZrO2), coated with slurry of mole ratio of SiO2to ZrO21:2 (group 1SiO2-2ZrO2), coated with slurry of mole ratio of SiO2to ZrO21:3 (group 1SiO2-3ZrO2), coated with slurry of mole ratio of SiO2to ZrO21:4 (group 1SiO2-4ZrO2). Profilometer, X-ray diffractometer (XRD), energy dispersive spectrometer, scanning electron microscopy (SEM) were used to analyze surface performance. RESULTS: The surface roughness of the discs in group AS was lower than those in the other groups [(0.33 ± 0.03) µm] (P < 0.05), there was no statistically significant difference (P > 0.05) among group 2SiO2-1ZrO2[(3.85 ± 0.38) µm], group 1SiO2-1ZrO2[(3.78 ± 0.56) µm] and group 1SiO2-2ZrO2[(4.06 ± 0.48) µm], and no difference (P > 0.05) was observed between group 1SiO2-3ZrO2[(1.02 ± 0.09) µm] and group 1SiO2-4ZrO2[(1.53 ± 0.23) µm] either. However, surface roughness in all coating groups was higher than those in group SB [(0.86 ± 0.05) µm] (P < 0.05). According to the XRD pattern, group AS and all coating groups consisted of 100% tetragonal airconia and monoclinic zirconia was detected at surface of group SB. Contents of surface silicon of coating groups increased significantly, however, no silicon was detected at sample surface of group AS and group SB. SEM showed that zirconia grains of coating exposed since part of silicon was etched by hydrofluoric acid, a three-dimensional network of intergrain nano-spaces was created. CONCLUSIONS: SiO2-ZrO2slurry coating could make surface of zirconia rough and increase Si content without creating monoclinic zirconia.

Induction of endotoxin tolerance by pathogenic Neisseria is correlated with the inflammatory potential of lipooligosaccharides and regulated by microRNA-146a.

In this article, we report that retreatment of human monocytic THP-1 cells and primary monocytes with pathogenic Neisseria or with purified lipooligosaccharides (LOS) after previous exposure to LOS induced immune tolerance, as evidenced by reduced TNF-α and IL-1ß cytokine expression. LOS that we have previously shown to vary in their potential to activate TLR4 signaling, which was correlated with differences in levels of lipid A phosphorylation, had similarly variable ability to induce tolerance. Efficacy for induction of tolerance was proportional to the level of lipid A phosphorylation, as LOS from meningococcal strain 89I with the highest degree of phosphorylation was the most tolerogenic following retreatment with LOS or whole bacteria, compared with LOS from gonococcal strains 1291 and GC56 with reduced levels of phosphorylation. Hydrogen fluoride treatment of 89I LOS to remove phosphates rendered the LOS nontolerogenic. Tolerance induced by the more highly inflammatory meningococcal LOS was correlated with significantly greater downregulation of p38 activation, greater induction of the expression of A20 and of microRNA-146a, and greater reductions in IL-1R-associated kinase 1 and TRAF6 levels following LOS retreatment of cells. The role of miR-146a in regulation of induction of TNF-α was confirmed by transfecting cells with an inhibitor and a mimic of miR-146a. Our results provide a mechanistic framework for understanding the variable pathophysiology of meningococcal and gonococcal infections given that after an initial exposure, greater upregulation of microRNA-146a by more highly inflammatory LOS conversely leads to the suppression of immune responses, which would be expected to facilitate bacterial survival and dissemination.

The effect of hydrofluoric acid treatment of titanium and titanium dioxide surface on primary human osteoblasts.

OBJECTIVE: The aim of the study was to investigate solely the effect of fluoride on the surface chemistry of polycrystalline ceramic titanium dioxide (TiO2 ) and metallic titanium (Ti) and its effect on proliferation and differentiation of primary human osteoblasts (NHO). MATERIALS AND METHODS: The NHO cells were exposed to fluoride-modified and unmodified samples for 1, 3, 7, 14 and 21 days. The fluoride effect on the mRNA expression was quantified and measured. The secretion of cytokines and interleukins in the cell culture medium was measured by Luminex, gene expression by RT-PCR, and compared with untreated controls. The effect on cell growth after 1 and 3 days in culture was measured using [(3) H]-thymidine incorporation. Fluoride release was measured using an ion-selective electrode. The surfaces were examined by X-ray photoelectron spectroscopy and profilometry. RESULTS: The fluoride release study detected that fluoride content easily washed off in TiO2 coins when compared with Ti coins. No increase in cell proliferation was found among fluoride-modified TiO2 surfaces compared with controls, except for washed Ti coins with fluoride modification. The cell differentiation with regard to gene expression showed no significant differences in both fluoride-modified and unmodified samples and less effect on protein release for all groups. CONCLUSIONS: The fluoride from hydrofluoric acid treatment on Ti and TiO2 surfaces gave no specific effect on primary human osteoblast cells. The study indicates that the released fluoride is not the unique factor for the bioactivity of Ti and TiO2 surfaces.

Comparison of the effects of surface treatments on roughness of two ceramic systems.

OBJECTIVE: The aim of this study was to evaluate the effects of different surface treatments on the surface roughness of lithium disilicate-based core (IPS Empress 2, shade 210, Ivoclar Vivadent, Schaan, Liechtenstein) and feldspathic ceramics (Vita VM9, VITA Zahnfabrik H. Rauter GmbH & Co. KG, Bad Säckingen, Germany). BACKGROUND DATA: Er:YAG laser irradiation is expected to be an alternative surface treatment, thus enhances surface roughness of procelains and produces morphological changes. METHODS: Fifty lithium disilicate-based core ceramic discs and 50 feldspathic ceramic discs were prepared (diameter, 10 mm; thickness, 1 mm) according to the manufacturers' instructions. All-ceramic discs were polished to standardize, and surface roughness of the discs was evaluated before treatment and serving as controls. Both of two ceramic groups were divided into five groups (n=10), and the following treatments were applied: (1) sandblasting with aluminum oxide (Al(2)O(3); Group SB); (2) Al(2)O(3)+Er:YAG laser (Group SB-L); (3) Er:YAG laser irradiation (distance, 1 mm; 500 mJ; 20 Hz; 10W; manually, contact handpiece [R 14]) (Group L); (4) 5% hydrofluoric acid etching (Group HF); and (5) Er:YAG laser +5% hydrofluoric acid (Group HF-L). Surface roughness was evaluated by profilometry, and specimens were then examined with atomic force microscopy. RESULTS: Data were analyzed by Wilcoxon signed rank, Mann-Whitney U, and Kruskal-Wallis tests (α=0.05). The Wilcoxon signed rank test results indicated that surface roughness after sandblasting was significantly different from the surface roughness after laser irradiation and acid etching (p<0.001). Mann-Whitney U and Kruskal-Wallis test results indicated that groups SB and SB-L had significantly higher mean roughness values (p<0.05) than those in the other groups. CONCLUSIONS: Groups SB and SB-L had rougher surfaces than the groups subjected to the other surface treatment methods. There was no significant difference in surface roughness between the HF acid etching, Er:YAG laser irradiation, and HF and Er:YAG (p<0 .05).

Inhibition assay of yeast cell walls by plasmon resonance Rayleigh scattering and surface-enhanced Raman scattering imaging.

We report on plasmon resonance Rayleigh scattering (PRRS) and surface enhanced Raman scattering (SERS) imaging for inhibition assay of yeast cell walls. This assay reveals that the proteins having alkali sensitive linkage bound to ß1,3 glucan frameworks in cell walls are involved in SERS activity. The result is further confirmed by comparison of genetically modified cells and wild type cells. Finally, we find that PRRS and SERS spots do not appear on cell walls when daughter cells are enough smaller than parent ones, but appear when size of daughter cells are comparable to parent cells. This finding indicates the relationship between expression of the proteins that generate SERS spots and cell division. These results demonstrate that PRRS and SERS imaging can be a convenient and sensitive method for analysis of cell walls.

Effects of hydrofluoric acid and anodised micro and micro/nano surface implants on early osseointegration in rats.

Our aim was to evaluate the effects of hydrofluoric acid and anodised micro and micro/nano surface implants on bony ingrowth in the earliest stage of implantation in rats. Sixty cylindrical screwed titanium alloy implants with machined, micro, and hierarchical hybrid micro/nano surfaces (n=20 in each group) were inserted into the distal femurs of 30 female Sprague-Dawley rats. In vivo microcomputed tomography (micro CT) was used to assess microarchitectural changes in the bone around the implants 2 weeks after implantation. All the animals were then killed and the femurs with implants harvested for histological analysis and pull-out testing. Micro CT analysis showed that the trabecular thickness and the bone:volume ratio (bone volume:total volume) (BV:TV) increased significantly in the micro/nano group compared with the other two groups, while the trabecular separation decreased significantly in the micro/nano group compared with the machined group. The mean (SD) bone-implant contacts (%) were 38.94 (9.48), 41.67 (8.71), and 51.49 (12.49) in the machined, micro, and micro/nano groups, respectively. The maximum pull-out forces (N) were 64.95 (6.11), 71.45 (7.15), and 81.90 (13.1), respectively. Both bone-implant contacts and maximum pull-out forces were significantly higher in the micro/nano group, but there was no significant difference between the micro group and the machined group. These data indicate that the hierarchical hybrid micro/nano surface of the implant can promote osseointegration in the earliest stage of implantation, and may be a promising option for further clinical use.

The silicon layer supports acid resistance of Bacillus cereus spores.

Silicon (Si) is considered to be a "quasiessential" element for most living organisms. However, silicate uptake in bacteria and its physiological functions have remained obscure. We observed that Si is deposited in a spore coat layer of nanometer-sized particles in Bacillus cereus and that the Si layer enhances acid resistance. The novel acid resistance of the spore mediated by Si encapsulation was also observed in other Bacillus strains, representing a general adaptation enhancing survival under acidic conditions.