Influence of antimicrobial peptide biofunctionalized TiO2 nanotubes on the biological behavior of human keratinocytes and its antibacterial effect / 中华口腔医学杂志

Objective: To fabricate TiO2 nanotube material functionalized by antimicrobial peptide LL-37, and to explore its effects on biological behaviors such as adhesion and migration of human keratinocytes (HaCaT) and its antibacterial properties. Methods: The TiO2 nanotube array (NT) was constructed on the surface of polished titanium (PT) by anodization, and the antimicrobial peptide LL-37 was loaded on the surface of TiO2 nanotube (LL-37/NT) by physical adsorption. Three samples were selected by simple random sampling in each group. Surface morphology, roughness, hydrophilicity and release characteristics of LL-37 of the samples were analyzed with a field emission scanning electron microscope, an atomic force microscope, a contact angle measuring device and a microplate absorbance reader. HaCaT cells were respectively cultured on the surface of three groups of titanium samples. Each group had 3 replicates. The morphology of cell was observed by field emission scanning electron microscope. The number of cell adhesion was observed by cellular immunofluorescence staining. Cell counting kit-8 (CCK-8) assay was used to detect cell proliferation. Wound scratch assay was used to observe the migration of HaCaT. The above experiments were used to evaluate the effect of each group on the biological behavior of HaCaT cells. To evaluate their antibacterial effects, Porphyromonas gingivalis (Pg) was respectively inoculated on the surface of three groups of titanium samples. Each group had 3 replicates. The morphology of bacteria was observed by field emission scanning electron microscope. Bacterial viability was determined by live/dead bacterial staining. Results: A uniform array of nanotubes could be seen on the surface of titanium samples in LL-37/NT group, and the top of the tube was covered with granular LL-37. Compared with PT group [the roughness was (2.30±0.18) nm, the contact angle was 71.8°±1.7°], the roughness [(20.40±3.10) and (19.10±4.11) nm] and hydrophilicity (the contact angles were 22.4°±3.1° and 25.3°±2.2°, respectively) of titanium samples increased in NT and LL-37/NT group (P<0.001). The results of in vitro release test showed that the release of antimicrobial peptide LL-37 was characterized by early sudden release (1-4 h) and long-term (1-7 d) slow release. With the immunofluorescence, more cell attachment was found on NT and LL-37/NT than that on PT at the first 0.5 and 2.0 h of culture (P<0.05). The results of CCK-8 showed that there was no significant difference in the proliferation of cells among groups at 1, 3 and 5 days after culture. Wound scratch assay showed that compared with PT and NT group, the cell moved fastest on the surface of titanium samples in LL-37/NT group at 24 h of culture [(96.4±4.9)%] (F=35.55, P<0.001). A monolayer cells could be formed and filled with the scratch in 24 h at LL-37/NT group. The results of bacterial test in vitro showed that compared with the PT group, the bacterial morphology in the NT and LL-37/NT groups was significantly wrinkled, and obvious bacterial rupture could be seen on the surface of titanium samples in LL-37/NT group. The results of bacteria staining showed that the green fluorescence intensity of titanium samples in LL-37/NT group was the lowest in all groups (F=66.54,P<0.001). Conclusions: LL-37/NT is beneficial to the adhesion and migration of HaCaT cells and has excellent antibacterial properties, this provides a new strategy for the optimal design of implant neck materials.

Research progress of tunneling nanotube in bone biology / 中华口腔医学杂志

Tunneling nanotube (TNT) is a newly discovered communication mode between animal cells in recent years, which have important physiological and pathological significance. However, the role of TNT in bone biology is still unclear. At present, there are many reports about tunneling nanotubes in bone marrow mesenchymal stem cells, osteoclast precursor cells, osteoblasts and immune cells. This review describes the research advances of TNT and its research progress in bone biology. It looks forward to the research direction of TNT in oral and maxillofacial bone development and bone biology, to provide new strategies for the maintenance of bone homeostasis and the treatment of bone diseases.

The effects of TiO2 nanotubes on bond strength and radiopacity of a self-adhesive resin cement in self-curing mode / Os efeitos de nanotubos de TiO2 na resistência de união e radiopacidade de um cimento resinoso auto-adesivo no modo auto-polimerizável

Objective: The aim of this in vitro study w as t o analyze the influence of the titanium dioxide nanotubes i n a self-cure mode polymerization of a dual resin luting agent through push out bond strength and radiopacity tests. Material and Methods: After mixed with a commercial du al self-adhesive resin cement, three concentrations o f tit anium dioxide nanotubes (0.3, 0.6, and 0.9% by weight) we re analyzed in self-curing mode. T he bond strength to bovine root dentin and fi berglass posts was assessed with the push out bond str ength t est and was evaluated in three thirds (cervical, middle and apical) (n=10), followed by failure mode analysis (SEM), and the ISO standard 9917-2 was followed for radiopacity test (n=10). Data were statistically analyzed by one-way ANOVA test, followed by Tukey's test (α=0.05). Results: Reinforced self-adhesive resin cement with 0.6% titanium dioxide nanotubes showed significant difference compared to the control gr oup for push out test (p=0.00158). The modified groups did not s how significant difference among thirds (p=0.782). Radiopacity sh owed higher value for group w ith 0.9% titanium dioxide nanotubes in comparison w ith control group (p<0.001). Conclusion: The addition of titanium dioxide nanotubes to a self-adhesive resin cement increased the bond strength to dentin and radiopacity values in the self-cure polymerization mode (AU)

Objetivo: O objetivo deste estudo in vitro foi analisar a influência de nanotubos de dióxido de titânio na polimerização química de um agente cimentante resinoso dual através de testes de resistência à união e radiopacidade. Material e Métodos: Após misturado com um cimento resinoso auto-adesivo comercial, três concentrações de nanotubos de dióxido de titânio (0,3, 0,6 e 0,9% em peso) foram analisadas. A resistência da união para a dentina da raiz bovina e os pinos de fibra de vidro foi avaliada pelo teste de push-out e avaliada em três terços (cervical, médio e apical) (n = 10), seguido pelo análise de modo de falha (MEV) e a norma ISO 9917-2 foi seguido para teste de radiopacidade (n = 10). Os dados foram analisados estatisticamente pelo teste ANOVA um fator seguido do teste de Tukey (α = 0,05). Resultados: O cimento resinoso auto-adesivo reforçado com nanotubos de dióxido de titânio a 0,6% mostrou diferença significativa em comparação com o grupo controle para teste de push-out (p=0,00158). Os grupos modificados não mostraram diferença significativa entre os terços (p=0,782). A radiopacidade mostrou maior valor para o grupo com nanotubos de dióxido de titânio 0,9% em comparação com o grupo controle (p<0,001). Conclusão: A adição de nanotubos de dióxido de titânio a um cimento resinoso auto-adesivo aumentou a os valores de resistência de união à dentina e radiopacidade no modo de polimerização química do agente cimentante (AU)

Titanium dioxide nanotubes added to glass ionomer cements affect S. mutans viability and mechanisms of virulence

Abstract This in vitro study evaluated the impact of TiO2 nanotubes (n-TiO2) incorporated into glass ionomer cement (GIC) on Streptococcus mutans (S. mutans) characteristics at cellular and molecular levels. n-TiO2, synthesized by the alkaline method (20 nm in size), was added to Ketac Molar EasyMix® at 0%, 3%, 5%, and 7% by weight. S. mutans strains were cultured on GIC disks with addition or not of n-TiO2 for 1, 3, and 7 days and the following parameters were assessed: inhibition halo (mm) (n=3/group); cell viability (live/dead) (n=5/group); cell morphology (SEM) (n=3/group); and gene expression by real-time PCR (vicR, covR, gtfB, gtfC, and gtfD) (n=6/group). The data were analyzed by the Kruskal-Wallis test, repeated-measures ANOVA or two-way ANOVA, and Tukey's and Dunn's post-hoc tests (α=0.05). The agar diffusion test showed a higher antibacterial property for 5% n-TiO2 compared with 3% and 7% (p<0.05) with no effect of time (1, 3, and 7 days). The cell number was significantly affected by all n-TiO2 groups, while viability was mostly affected by 3% and 5% n-TiO2, which also affected cell morphology and organization. Real-time PCR demonstrated that n-TiO2 reduced the expression of covR when compared with GIC with no n-TiO2 (p<0.05), with no effect of time, except for 3% n-TiO2 on vicR expression. Within-group and between-group analyses revealed n-TiO2 did not affect mRNA levels of gtfB, gtfC, and gtfD (p>0.05). Incorporation of n-TiO2 at 3% and 5% potentially affected S. mutans viability and the expression of key genes for bacterial survival and growth, improving the anticariogenic properties of GIC.

Biological efficacy of perpendicular type-I collagen protruded from TiO / 国际口腔科学杂志·英文版

The aim of this study was to evaluate the biological efficacy of a unique perpendicular protrusion of type-I collagen (Col-I) from TiO

Titanium dioxide nanotubes incorporated into bleaching agents: physicochemical characterization and enamel color change

Abstract Titanium dioxide nanotubes are nanostructures that can accelerate the oxidation reaction of bleaching procedures and promote a more effective whitening effect. Objective This study evaluated physicochemical properties of bleaching agents incorporated with titanium dioxide (TiO2) nanotubes, and the effects on tooth color change at different periods. Methodology 40 premolars were treated according to the following groups (n=10): CP - 10% carbamide peroxide (1 hour daily/21 days); CPN - CP incorporated into TiO2; HP - 40% hydrogen peroxide (three 40-minute sessions/7 days apart); HPN - HP incorporated into TiO2. Color shade was evaluated at five different periods (baseline, after 7, 14 and 21 days of bleaching, and 7 days after end of treatment) according to Vita Classical, CIELab and CIEDE2000 scales. Mean particle size (P), polydispersity (PO) and zeta potential (ZP) were evaluated using dynamic light scattering. Data on the different variables were analyzed by mixed model tests for measures repeated in time (ZP e L*), generalized linear models for measures repeated in time (P, PO, Vita Classical and b*), and Friedman and Mann-Whitney tests (a* and color change/ΔE and ΔE00). Results CP and CPN presented higher P, higher PO and lower ZP than HP and HPN (p≤0.05). All groups showed a significant decrease in Vita Classical color scores after 7 days of bleaching (p<0.05), and HPN presented a greater significant reduction than the other groups. L* increased in TiO2 presence, in all groups, without any differences (p>0.05) in bleaching time. A significant reduction occurred in the a* and b* values for all the groups, and HPN presented lower a* and b* values (p<0.05) than CPN. ΔE was clinically noticeable after 7 days, in all groups, and all groups resulted in a perceptible color change according to ΔE00. Conclusion TiO2 did not influence physicochemical properties of the bleaching agents. HPN presented more effective tooth bleaching than CPN.

The biological functions of cell-to-cell connection over long distance--membrane nanotube / 生理学报

Cell-to-cell connections provide conduits for signal exchanges, and play important functional roles in physiological and pathological processes of multicellular organisms. Membrane nanotubes are common long-distance connections between cells, not only transfer molecule signals and mitochondria, but also cooperate with gap junction and other cell-to-cell communications to transfer signals. During the last decade, there are many studies about membrane nanotubes, which focus on the similarities and differences between membrane nanotubes and other cell-to-cell communications, as well as their biological functions. In the present review, we summarized the latest findings about the structural diversity, the similarities and differences in signal transmission with other types of cell-to-cell communications, and physiological and pathological roles of membrane nanotubes.

Differentiation Capacity of Monocyte-Derived Multipotential Cells on Nanocomposite Poly(e-caprolactone)-Based Thin Films

BACKGROUND: Lonocyte-derived multipotential cells (MOMCs) include progenitors capable of differentiation into multiple cell lineages and thus represent an ideal autologous transplantable cell source for regenerative medicine. In this study, we cultured MOMCs, generated from mononuclear cells of peripheral blood, on the surface of nanocomposite thin films. METHODS: For this purpose, nanocomposite Poly(e-caprolactone) (PCL)-based thin films containing either 2.5 wt% silica nanotubes (SiO2ntbs) or strontium hydroxyapatite nanorods (SrHAnrds), were prepared using the spin-coating method. The induced differentiation capacity of MOMCs, towards bone and endothelium, was estimated using flow cytometry, real-time polymerase chain reaction, scanning electron microscopy and fluorescence microscopy after cells' genetic modification using the Sleeping Beauty Transposon System aiming their observation onto the scaffolds. Moreover, Wharton's Jelly Mesenchymal Stromal Cells were cultivated as a control cell line, while Human Umbilical Vein Endothelial Cells were used to strengthen and accelerate the differentiation procedure in semi-permeable culture systems. Finally, the cytotoxicity of the studied materials was checked with MTT assay. RESULTS: The highest differentiation capacity of MOMCs was observed on PCL/SiO2ntbs 2.5 wt% nanocomposite film, as they progressively lost their native markers and gained endothelial lineage, in both protein and transcriptional level. In addition, the presence of SrHAnrds in the PCL matrix triggered processes related to osteoblast bone formation. CONCLUSION: To conclude, the differentiation of MOMCs was selectively guided by incorporating SiO2ntbs or SrHAnrds into a polymeric matrix, for the first time.

Design and characterization of nanocarriers loaded with Levofloxacin for enhanced antimicrobial activity; physicochemical properties, in vitro release and oral acute toxicity

Inorganic and carbon based nanomaterials are widely used against several diseases, such as cancer, autoimmune diseases as well as fungi and bacteria colonization. In this work, Santa Barbara Amorphous mesoporous silica (SBA), Halloysite Nanotubes (HNTs) and Multiwalled Carbon Nanotubes (CNTs) were loaded with fluoroquinolone Levofloxacin (LVF) to be applied as antimicrobial agents. The prepared via adsorption nanocarriers were characterized by Fourier-Transformed Spectroscopy, Scanning Electron Microscopy as well as High Pressure liquid Chromatography. In vitro release studies were carried out using Simulated Body Fluid at 37oC and data analyzed by various kinetic models showing slow dissolution over 12-24 hours. Antimicrobial studies showed improved antibacterial activity against Escherichia coli, Enterococcus faecalis, Listeria monocytogenes, Staphylococcus aureus, and Staphylococcus epidermidis compared to neat nanomaterials. CNTs were found to be the most promising candidates for LVF delivery and they were chosen to be further studied for their acute oral toxicity and histopathological examination using C57/Black mice. Histological examination depicted that drug loading did not affect mice organs morphology as well as hepatocyte degeneration, central vein degeneration and parenchymal necrosis scores. To conclude, the prepared nanomaterials present significant characteristics and can act as antimicrobial drug carriers; CNTs found to be safe candidates when orally fed to mice.

Evaluation of bioactivity and osseointegration for ti-6al-4v alloy implant modified by anodic oxidation and cyclic precalcification treatments / 대한치과재료학회지

The aim of this study was to examine the bioactivity and osseointegration of Ti-6Al-4V alloy implant which was modified by an anodic oxidation and a cyclic precalcification treatments. After blasting treatment using HAp (Hydroxyapatitie; HAp) powder which is resorbable blasting media (RBM) on the surface of Ti-6Al-4V alloy implants, the anodic oxidation treatment and the cyclic precalcification treatment were conducted to form nanotube TiO2 layer and HAp precipitation respectively. The surface morphology of the surface-treated Ti-6Al-4V alloy implant was investigated after immersion in the simulated body fluid(SBF) for 3 days to investigate the bioactivity. To investigate the effect of surface treatment on bonding between the implant and bone, RBM treated implant and RBM-anodization-cyclic precalcification(RACP) treated implant were placed on the distal side of both tibia diaphysis of rats, and then the removal torque of the implant was measured after 4 weeks. On the surface of RACP treated group, bone-like apatite precipitation was observed after immersion in SBF for 3 days. The removal torque was significantly higher in the RACP treated group than in the RBM treated group. The interfacial fracture between the implant and the new bone was observed in the RBM treated group, but both the cohesive fracture at the new bone and the interfacial fractures between the implant and the new bone were observed in the RACP treated group.

Effects of conditions for anodization and cyclic precalcification treatments on surface characteristics and bioactivity / 대한치과재료학회지

The purpose of this study was to investigate the effects of the anodization and cyclic calcification treatment on the surface characteristic and bioactivity of the titanium thin sheet in order to obtain basic data for the production of bioactive titanium membrane. A 30×20×0.08 mm titanium sheets were prepared, and then they were pickled for 10 seconds in the solution which was mixed with HNO₃: HF: H₂O in a ratio of 12: 7: 81. The TiO₂ nanotube layer was formed to increase the specific surface area of the titanium, and then the cyclic calcification treatment was performed to induce precipitation of hydroxiapatite by improvement of the bioactivity. The corrosion resistance test, wettability test and immersion test in simulated body solution were conducted to investigate the effect of these surface treatments. The nanotubes formed by the anodization treatment have a dense structure in which small diameter tubes are formed between relatively large diameter tubes, and their inside was hollow and the outer walls were coupled to each other. The hydroxyapatite precipitates were well combined on the nanotubes by the penetration into the nanotube layer by successive cyclic calcification treatment, and the precipitation of hydroxyapatite tended to increase proportionally after immersion in simulated body solution as the number of cycles increased. In conclusion, it was confirmed that induction of precipitation of hydroxyapatite by cyclic calcification treatment after forming the nanotube TiO₂ nanotube layer on the surface of the titanium membrane can contribute to improvement of bioactivity.

Titania nanotube template based synthesis of gold nanotubes and their antibacterial activity / 대한치과재료학회지

In this study, gold nanotubes were fabricated by electrophoretic deposition using a titania nanotube layer as a template, and then the surface characteristics, biocompatibility and antibacterial effect of gold nanotubes were evaluated. Gold nanotubes of 100 nm diameter were fabricated by depositing 4 nm and 15 nm gold nanoparticles on anodized 100 nm titania nanotubes by citrate reduction and electrophoretic deposition. As a result of the UV-Vis diffuse spectrophotometer, 4 nm and 15 nm gold nanotubes showed strong absorption at 702~774 nm and 753~760 nm, respectively. Also, the maximum absorption wavelength was shifted to the longer wavelength as the coating time of the gold nanoparticles increased. FE-SEM observation and EDX analysis resulted that 0.1~0.5 wt% gold nanoparticles uniformly were stacked on the top layer of titania nanotubes. As a result of MTT cell test, the relative absorbance value of all experimental groups after 24 hours and 48 hours of incubation exceeded 70% indicating excellent biocompatibility. The effect of the near infrared laser light on the adhesion and growth of gold nanotubes showed excellent antibacterial activity regardless of the coating time of gold nanoparticles. Therefore, it is confirmed that the gold nanotube coating technology based on the titania nanotube template is supposed to be highly applicable to a titanium implant surface treatment technology with the remote control thermal treatment of a near-infrared laser.

Effects of incorporation of 2.5 and 5 wt% TiO₂ nanotubes on fracture toughness, flexural strength, and microhardness of denture base poly methyl methacrylate (PMMA)

PURPOSE: The aim of this preliminary study was to investigate, for the first time, the effects of addition of titania nanotubes (n-TiO2) to poly methyl methacrylate (PMMA) on mechanical properties of PMMA denture base. MATERIALS AND METHODS: TiO2 nanotubes were prepared using alkaline hydrothermal process. Obtained nanotubes were assessed using FESEM-EDX, XRD, and FT-IR. For 3 experiments of this study (fracture toughness, three-point bending flexural strength, and Vickers microhardness), 135 specimens were prepared according to ISO 20795-1:2013 (n of each experiment=45). For each experiment, PMMA was mixed with 0% (control), 2.5 wt%, and 5 wt% nanotubes. From each TiO2:PMMA ratio, 15 specimens were fabricated for each experiment. Effects of n-TiO2 addition on 3 mechanical properties were assessed using Pearson, ANOVA, and Tukey tests. RESULTS: SEM images of n-TiO2 exhibited the presence of elongated tubular structures. The XRD pattern of synthesized n-TiO2 represented the anatase crystal phase of TiO2. Moderate to very strong significant positive correlations were observed between the concentration of n-TiO2 and each of the 3 physicomechanical properties of PMMA (Pearson's P value ≤.001, correlation coefficient ranging between 0.5 and 0.9). Flexural strength and hardness values of specimens modified with both 2.5 and 5 wt% n-TiO2 were significantly higher than those of control (P≤.001). Fracture toughness of samples reinforced with 5 wt% n-TiO2 (but not those of 2.5% n-TiO2) was higher than control (P=.002). CONCLUSION: Titania nanotubes were successfully introduced for the first time as a means of enhancing the hardness, flexural strength, and fracture toughness of denture base PMMA.

Hydroxyapatite Nanorod-Modified Sand Blasted Titanium Disk for Endosseous Dental Implant Applications

BACKGROUND: Sand blasted titanium (Ti) is commonly used in designing endosseous dental implants due to its biocompatibility and ability to form bonds with bone tissues. However, titanium implants do not induce strong interactions with teeth bones. To increase strong interactions between Ti disk implants and teeth bones, the L-glutamic acid grafted hydroxyapatite nanorods (nHA) were immobilized on albumin modified Ti disk implants (Ti-Alb). METHODS: For modification of Ti disk implants by nHA, the L-glutamic acid grafted nHA was synthesized and then immobilized on albumin modified Ti disk implants. Fourier transformed infrared spectroscopy, X-ray photoelectron spectroscopy, scanning electron microscope; energy dispersive spectroscopy and confocal laser scanning microscopy were used to confirm the modification of Ti disk implants. The bioactivity of nHA-modified Ti disk implants was evaluated by seeding MC3T3-E1 cells on Ti-nHA implants. RESULTS: Characterization techniques have confirmed the successful modification of Ti disk implants by L-glutamic acid grafted nHA. The nHA-modified Ti disk implants have shown enhanced adhesion, proliferation and cytotoxicity of MC3T3-E1 cells in comparison to pristine Ti implants. CONCLUSION: The modification of Ti implants by L-glutamic acid grafted nHA has produced highly osteogenic Ti disk plants in comparison to pristine Ti disk implants due to the formation of bioactive surfaces by hydroxyapatite nano rods on Ti disk implants. Ti-nHA disk implants showed enhanced adhesion, proliferation, and MC3T3-E1 cells viability in comparison to pristine Ti disk implants. Thus nHA might be to be useful to enhance the osseointegration of Ti implants with teeth bones.

A study of the modification of Y-TZP and resin cement with titanium dioxide nanotubes / Estudo da modificação de Y-TZP e cimento resinoso com nanotubos de dióxido de titânio

Titanium dioxide nanotubes (TiO2) have been applied to enhance the mechanical properties of dental materials. Yttria-stabilized tetragonal zirconia polycrystals (Y-TZP) has been increasingly used in dentistry. Aside from its optimal clinical results, Y-TZP is prone to failures related to insufficient thickness of the fixed prostheses connector and debonding due to its difficult adhesion. The purpose of this study is to evaluate the effect of the addition of different concentrations of TiO2 to YTZP in its mechanical properties and microstructure, and also to evaluate the influence of these nanotubes on the bond strength when added to the ceramic or to the resin cement. To evaluate that, the described purposes were divided in two different papers. Paper 1 describes the tests of biaxial flexural strength, fractography qualitative analysis in scanning electron microscopy (SEM), microstructure evaluation in field emission-SEM and X-ray diffraction. Groups evaluated were commercial YTZP (Ivoclar Vivadent) (ZC) and an experimental Y-TZP with different blends of nanotubes [0 (Z0), 1 (Z1), 2 (Z2), and 5% (Z5), in volume]. In paper 2 shear bond strength test is described. It was carried out with the following groups: commercial YTZP (Ivoclar Vivadent) (ZC) and an experimental Y-TZP with different blends of nanotubes [0 (Z0), 1 (Z1), 2 (Z2), and 5% (Z5), in volume] bonded to the resin cement Panavia F2.0; and commercial Y-TZP bonded to resin cement RelyX U200 added with different blends of nanotubes in two curing methods dual-cured [0 (DC), 0.3 (D03), 0.6 (D06) and 0.9% (D09) of nanotubes in weigth] or self-cured [0 (SC), 0.3 (S03), 0.6 (S06) and 0.9% (S09) of nanotubes in weigth]. Values of flexural strength and shear bond strength were subjected to ANOVA and Tukey (=0.05). Flexural strength values were also subjected to Weibull statistics. Grain sizes values were subjected to Kruskal-Wallis and Dunn tests (=0.05). The flexural strength results were: ZC 896.73±122.70; Z0 577.67±62.26; Z1 477.32±75.65; Z2 492.25±63.19; Z5 437.18±53.55. The Weibull modulus results found were: ZC 7.9; Z0 11.2, Z1 8.7; Z2 8.1; Z5 9.3. Results showed that experimental Y-TZP presented lower flexural strength values than commercial one, but the first presented better Weibull modulus (m). Experimental Y-TZP also presented good microstructure, comparable to commercial Y-TZP, with very similar grain sizes. Nanotubes addition to Y-TZP led to lower flexural strength, although higher m than commercial ceramic. Pores containing Ti were observed in Y-TZP as the nanotubes concentration raised. Shear bond strength results found were, from higher to lower values: Z5 6.46±3.36; DC 6.17±0.87; D03 5.74±1.70; S03 5.73±1.71; Z1 5.16±2.62; D06 4.82±1.06; D09 4.75±1.43; SC 4.73±1.43; S09 4.61±0.85; S06 4.51±1.87; ZC 3.70±1.82; Z0 -3.33±2.05; Z2 2.94±1.38. Shear bond strength was also influenced by nanotubes addition, either in the ceramic or in the cement, although not linearly. Y-TZP added of 5% of TiO2 nanotubes presented the highest bond strength, although with no significant difference from most groups. Group Z1 was probably the group that presented the best combination of flexural strength, m, microstructure and bond strength. More studies of other properties could be carried out with this group.(AU)

Nanotubos de dióxido de titânio (TiO2) tem sido utilizados para melhorar as propriedades mecânicas de materiais odontológicos. Zircônica tetragonal policristalina estabilizada por ítria (Y-TZP) tem sido amplamente utilizada na Odontologia. Apesar de seus excelentes resultados clínicos, a Y-TZP é suscetível a falhas relacionadas à espessura insuficiente do conector da prótese fixa e a soltura da restauração devido à adesão deficiente. O objetivo desse trabalho é avaliar o efeito da adição de diferentes concentrações de nanotubos de TiO2 à Y-TZP nas suas propriedades mecânicas e microestrutura, além de avaliar a influência na resistência de união da adição desses nanotubos na cerâmica ou no cimento resinoso. Para isso, os objetivos descritos foram divididos em dois artigos diferentes. O artigo 1 descreve os testes de resistência flexural biaxial, análise fractográfica qualitativa em microscopia eletrônica de varredura (MEV), avaliação de microestrutura em MEV de emissão de campo e difração de raios-X. Os grupos avaliados foram: Y-TZP comercial (Ivoclar Vivadent) (ZC) e Y-TZP experimental com diferentes concentrações de nanotubos [0 (Z0), 1 (Z1), 2 (Z2), e 5% (Z5), em volume]. No artigo 2 está descrito o teste de resistência ao cisalhamento que foi realizado com os seguintes grupos: Y-TZP comercial (ZC) e Y-TZP experimental com diferentes concentrações de nanotubos [0 (Z0), 1 (Z1), 2 (Z2), e 5% (Z5), em volume] aderidas ao cimento Panavia F2.0; e Y-TZP comercial aderida ao cimento resinoso RelyX U200 com adição de diferentes concentrações de nanotubos em dois métodos de polimerização: dual [0 (DC), 0,3 (D03), 0,6 (D06) e 0,9% (D09) de nanotubos em peso] ou auto [0 (SC), 0,3 (S03), 0,6 (S06) e 0,9% (S09) de nanotubos em peso]. Os valores de resistência flexural e resistência ao cisalhamento foram submetidos aos testes de ANOVA e Tukey (=0,05). A resistência flexural também passou por análise de Weibull. Os valores de tamanho de grãos foram submetidos a testes de Kruskal-Wallis e Dunn (=0,05). Os resultados de resistência flexural encontrados foram: ZC 896,73±122,70; Z0 577,67±62,26; Z1 477,32±75,65; Z2 492,25±63,19; Z5 437,18±53,55. Os resultados de módulo de Weibull encontrados foram: ZC - 7,9; Z0 - 11,2; Z1 - 8,7; Z2 - 8,1; Z5 - 9,3. Os resultados mostraram que a Y-TZP experimental apresentou menores valores de resistência flexural do que a cerâmica comercial, mas a primeira apresentou maior módulo de Weibull (m). A Y-TZP experimental apresentou boa microestrutura, comparável à Y-TZP comercial, com tamanhos de grão muito semelhantes. A adição de nanotubos à Y-TZP levou a menor resistência flexural, porém maior m que a cerâmica comercial. Poros contendo Ti foram observados na Y-TZP conforme a concentração de nanotubos aumentou. Os resultados de resistência ao cisalhamento foram, do maior para o menor: Z5 - 6,46±3,36; DC - 6,17±0,87; D03 - 5,74±1,70; S03 - 5,73±1,71; Z1 - 5,16±2,62; D06 - 4,82±1,06; D09 - 4,75±1,43; SC - 4,73±1,43; S09 - 4,61±0,85; S06 - 4,51±1,87; ZC - 3,70±1,82; Z0 - 3,33±2,05; Z2 - 2,94±1,38. A resistência ao cisalhamento também foi influenciada pela adição de nanotubos, tanto na cerâmica quanto no cimento, porém não linearmente. A Y-TZP adicionada de 5% de nanotubos de TiO2 apresentou maior resistência de união, porém sem diferença estatística da maioria dos grupos. O grupo Z1 foi provavelmente o grupo que apresentou a melhor combinação de resistência flexural, m, microestrutura e resistência de união. Mais estudos de outras propriedades podem ser realizados com o mesmo.(AU)

Electrochemical Characteristics of Nanotubular Ti-25Nb-xZr Ternary Alloys for Dental Implant Materials

PURPOSE: The purpose of this study was to investigate the electrochemical characteristics of nanotubular Ti-25Nb-xZr ternary alloys for dental implant materials. MATERIALS AND METHODS: Ti-25Nb-xZr alloys with different Zr contents (0, 3, 7, and 15 wt.%) were manufactured using commercially pure titanium (CP-Ti), niobium (Nb), and zirconium (Zr) (99.95 wt.% purity). The alloys were prepared by arc melting in argon (Ar) atmosphere. The Ti-25Nb-xZr alloys were homogenized in Ar atmosphere at 1,000℃ for 12 hours followed by quenching into ice water. The microstructure of the Ti-25Nb-xZr alloys was examined by a field emission scanning electron microscope. The phases in the alloys were identified by an X-ray diffractometer. The chemical composition of the nanotube-formed surfaces was determined by energy-dispersive X-ray spectroscopy. Selforganized TiO₂ was prepared by electrochemical oxidation of the samples in a 1.0 M H₃PO₄+0.8 wt.% NaF electrolyte. The anodization potential was 30 V and time was 1 hour by DC supplier. Surface wettability was evaluated for both the metallographically polished and nanotube-formed surfaces using a contact-angle goniometer. The corrosion properties of the specimens were investigated using a 0.9 wt.% aqueous solution of NaCl at 36℃±5℃ using a potentiodynamic polarization test. RESULT: Needle-like structure of Ti-25Nb-xZr alloys was transform to equiaxed structure as Zr content increased. Nanotube formed on Ti-25Nb-xZr alloys show two sizes of nanotube structure. The diameters of the large tubes decreased and small tubes increased as Zr content increased. The lower contact angles for nanotube formed Ti-25NbxZr alloys surfaces showed compare to non-nanotube formed surface. The corrosion resistance of alloy increased as Zr content increased, and nanotube formed surface showed longer the passive regions compared to non-treatment surface. CONCLUSION: It is confirmed that corrosion resistance of alloy increased as Zr content increased, and nanotube formed surface has longer passive region compared to without treatment surface.

Caracterização físico-química, mecânica e biológica de cimento autoadesivo aditivado com nanotubos de dióxido de titânio / Physical-chemical, mechanical and biological characterization of reinforcement self-adhesive resin cement titanium dioxide nanotubes

O objetivo desse estudo foi realizar a caracterização do cimento resinoso autoadesivo RelyX U200 aditivado de nanotubos de dióxido de titânio (nt-TiO2) em diferentes concentrações (0,3, 0,6, e 0,9% em peso) quanto às suas propriedades físico-químicas, mecânicas e biológica. Duas condições de polimerização foram analisadas: autopolimerizável (grupos: AC, A03, A06 e A09) e dual (grupos: DC, D03, D06 e D09). Para análise do grau de conversão foi utilizada a espectroscopia de infravermelho com transformada de Fourier com registro do espectro nos tempos de 3, 6, 9, 12 e 15 minutos. Os picos das bandas de comprimento de onda de 1638 cm-1 e de 1608 cm-1 foram identificados para cálculo do grau de conversão. Os dados foram submetidos à ANOVA de medidas repetidas seguido de comparações múltiplas de Tukey (=0,05). A análise de sorção e solubilidade foi realizada por meio da confecção de discos de cimento resinoso (10 mm ø × 2 mm) (n=8) monitorados quanto à sua massa depois de ciclos de hidratação/desidratação. A resistência à flexão em 3 pontos e módulo de elasticidade foram mensurados por meio de barras (2 × 2 × 25 mm) de cimento resinoso (n=10) levadas à máquina universal de ensaios. Para análise de Dureza Knoop utilizou-se microdurômetro com carga de 50g /10 segundos. Nos discos de cimento resinoso de 10 mm ø × 2 mm foram realizadas 5 endentações equidistantes 0,5mm e medidas em aumento de 50×. Para resistência de união ao cisalhamento, sobre discos de zircônia sinterizados foi aplicado o cimento resinoso (n=10) nas dimensões de 3 mm ø × 2 mm. Por meio de dispositivo foram levados à máquina universal de ensaios. Os dados encontrados de sorção e solubilidade e de cada propriedade mecânica foram submetidos aos testes ANOVA a dois critérios e de comparações múltiplas de Tukey (=0,05). Exceto para resistência ao cisalhamento que se utilizou o teste de comparação de Fischer (=0,05). Para viabilidade celular (n=8) foi realizado teste de MTT apenas na condição dual. Os grupos estudados foram: DC, D03, D06, D09, CP (controle positivo), CN (controle negativo). Após 24, 48 e 72 horas os níveis de absorbâncias foram analisados por meio de espectrofotometria no leitor de ELISA. Os dados foram submetidos aos testes ANOVA a dois critérios e de comparações múltiplas de Tukey (=0,05). Os resultados mostraram que a adição de nt-TiO2, independente da concentração, aumentou os valores de grau de conversão do cimento resinoso para a condição autopolimerizável e dual em todos os tempos estudados. Já para sorção e solubilidade não houve influência nos resultados da concentração de nanotubos inseridos e da condição de polimerização. Para resistência flexural, a adição de nt-TiO2 nas concentrações de 0,3% (A03) e 0,9% (A09) resultou em dados similares ao controle na condição dual (DC). O valor médio de módulo de elasticidade aumentou com a adição de 0,9% (A09), similar a todos os grupos da condição dual, em que adição de nt-TiO2 não influenciou os resultados. A adição de 0,6% (A06 e D06) e 0,9% (A09 e D09) de nt-TiO2 ao cimento aumentou os valores de dureza quando comparado aos grupos controle (AC e DC). Para resistência de união ao cisalhamento, a concentração de 0,3% de nt-TiO2 (A03 e D03) aumentou os valores quando comparado aos grupos A06, D06, A09 e D09 porém sem diferença para os grupos controle (AC e DC). Para viabilidade celular no período de 24h, os grupos D03, D06 e D09 obtiveram resultado similar ao grupo CP, já o grupo DC apresentou valores de absorbância inferiores ao CP, usado como parâmetro de comparação. Em 48 e 72h, todos os grupos experimentais não demonstraram diferença significativa em comparação ao grupo CP. O grupo CN apresentou diferença para os demais em todos dos tempos estudados. A adição de nt-TiO2 ao cimento resinoso autoadesivo representa uma estratégia promissora para potencializar suas propriedades físico-químicas e mecânicas sem prejuízo das propriedades biológicas.(AU)

The aim of this study was to characterize the self-adhesive resin cement RelyX U200 additive, titanium dioxide nanotubes (nt-TiO2), at different concentrations (0.3%, 0.6%, and 0.9% by weight) and to determine their physicochemical, mechanical and biological properties. Two polymerization conditions were analyzed: self-curing (groups AC, A03, A06 and A09) and dual-curing (groups DC, D03, D06 and D09). To analyze the conversion degree, Fourier transform infrared spectroscopy was used, and the spectrum was recorded at 3, 6, 9, 12 and 15 minutes. The peaks of the wavelength bands, 1638 cm-1 and 1608 cm-1, were identified to calculate the degree of conversion. The data were subjected to a repeated-measures ANOVA followed by a Tukey multiple-comparison test (=0.05). The sorption and solubility analysis was performed by making resin-cement discs (10 mm ø × 2 mm) (n=8) and monitoring their masses after the hydration/dehydration cycles. The 3-point flexural strength and the modulus of elasticity of resin-cement bars (2 × 2 × 25 mm) (n=10) were measured using a universal testing machine. The Knoop microhardness was analyzed with a load of 50 g and a time of 10 seconds. On each resin cement disc, 5 equidistant indentations of 0.5 mm were made, and the measures were increased by 50×. To test bonding shear strength, resin cement was applied to sintered zirconia discs (3 mm ø × 2 mm) (n=10). The bonded discs were then taken to the universal testing machine. Their sorption, solubility and each mechanical property were submitted to a two-way ANOVA and a Tukey multiple-comparison test (=0.05). The shear strength was submitted to a Fischer comparison test (=0.05). To test cell viability (n=8), a MTT assay was performed using only the dual-curing condition. The studied groups were: DC, D03, D06, D09, CP (positive control) and CN (negative control). After 24, 48 and 72 hours, the absorbance levels were analyzed using an ELISA spectrophotometry reader. The data were submitted to a two-way ANOVA and a Tukey multiple-comparison test (=0.05). The results showed that the addition of nt-TiO2, regardless of concentration, increased the conversion degree values for the self-curing resin cement and for the dual-curing at all times studied. The sorption and solubility were not influenced by the concentration of the nanotubes or the polymerization condition. Regarding flexural strength, the addition of the nt-TiO2 in concentrations of either 0.3% (A03) or 0.9% (A09) resulted in data similar to those in the dual-curing control (DC) condition. The average modulus of elasticity increased with the addition of 0.9% nt-TiO2 (A09), and as with all the groups in the dual-curing condition, the addition of nt-TiO2 did not affect the results. The addition of either 0.6% (A06 and D06) or 0.9% (A09 and D09) of nt-TiO2 cement increased hardness values relative to the control groups (AC and DC). The group with a 0.3% concentration of nt-TiO2 (A03 and D03) showed higher bonding shear strength values than several of the groups with higher concentrations (A06, D06 and D09), but the A09 group had no difference relative to either control group (AC or DC). For cell viability in the 24-h period, the D03 D06 and D09 groups achieved a result similar to that of the CP group with significant difference to the DC group that had lower absorbance values. At the benchmarks of 48 and 72 h, and only the CN group showed a significant difference compared to others. The addition of nt-TiO2 is a promising strategy for improving the physical-chemical and mechanical properties without prejudice the biological properties.(AU)

Evaluation of polymethyl methacrylate resin mechanical properties with incorporated halloysite nanotubes

PURPOSE: This study inspects the effect of incorporating halloysite nanotubes (HNTs) into polymethyl methacrylate (PMMA) resin on its flexural strength, hardness, and Young's modulus. MATERIALS AND METHODS: Four groups of acrylic resin powder were prepared. One group without HNTs was used as a control group and the other three groups contained 0.3, 0.6 and 0.9 wt% HNTs. For each one, flexural strength, Young's modulus and hardness values were measured. One-way ANOVA and Tukey's test were used for comparison (P<.05). RESULTS: At lower concentration (0.3 wt%) of HNT, there was a significant increase of hardness values but no significant increase in both flexural strength and Young's modulus values of PMMA resin. In contrast, at higher concentration (0.6 and 0.9 wt%), there was a significant decrease in hardness values but no significant decrease in flexural strength and Young's modulus values compared to those of the control group. CONCLUSION: Addition of lower concentration of halloysite nanotubes to denture base materials could improve some of their mechanical properties. Improving the mechanical properties of acrylic resin base material could increase the patient satisfaction.

Prevalência de angina do peito pelo questionário de Rose na população brasileira: análise da Pesquisa Nacional de Saúde, 2013 / Prevalence of angina pectoris in the Brazilian population from the Rose questionnaire: analysis of the National Health Survey, 2013

RESUMO: Objetivo: Estimar a prevalência de angina do peito na população adulta brasileira com a aplicação do questionário de Rose para angina na Pesquisa Nacional de Saúde (PNS-2013). Métodos: Inquérito populacional representativo da população brasileira acima de 18 anos de idade, com amostragem probabilística conglomerada em três estágios. Foram obtidos registros de entrevistas de 60.202 indivíduos no território nacional. Apresentou-se ao entrevistado o questionário de Rose curto com três questões, adaptado por Lawlor em 2003 e validado no Brasil, para identificar angina do peito grau I (leve) e II (moderada/grave). Calcularam-se os valores de prevalência com intervalo de confiança de 95% (IC95%) segundo sexo, faixa etária, escolaridade e raça/cor. Resultados: A prevalência de angina leve (grau I) foi de 7,6% (IC95% 7,2 - 8,0) para toda população, com frequência maior em mulheres - 9,1% (IC95% 8,5 - 9,7) - do que em homens - 5,9% (5,3 - 6,4). A frequência de angina moderada/grave (grau II) foi 4,2 (IC95% 3,9 - 4,5), também mais frequente em mulheres - 5,2% (IC95% 4,7 - 5,6) - do que em homens - 3,0% (IC95% 2,7 - 3,4). A prevalência de angina por faixa etária aumentou progressivamente com a idade. A prevalência de angina, de qualquer tipo, foi inversa aos anos de estudo formal. Apesar do valor maior da presença de angina em negros, não houve diferença significativa por raça/cor da pele. Conclusão: Os valores de prevalência elevada de angina do peito na população brasileira acima de 18 anos foram compatíveis com estudos em outros países, revelando a importância da doença coronariana como problema de saúde pública.

ABSTRACT: Objective: To estimate the prevalence of angina pectoris in the Brazilian adult population with the use of the Rose questionnaire for angina in the National Health Survey (PNS 2013). Methods: Population survey representing the Brazilian population aged 18 years and older, with probability carried out sampling in three stages. The interview records of 60,202 individuals were obtained in the country. The respondent was presented with the short Rose questionnaire with three questions, adapted by Lawlor in 2003 and validated in Brazil, to identify angina pectoris grade I (mild) and II (moderate/severe). The prevalence rate was calculated with a 95% confidence interval (95%CI) according to sex, age, education, and race/color. Results: The prevalence of mild angina (grade I) was of 7.6% (95%CI 7.2 - 8.0) for the entire population, more frequently in women - 9.1% (95%CI 8.5 - 9.7) - than in men - 5.9% (95%CI 5.3 - 6.4). The frequency of moderate/severe angina (grade II) was of 4.2 (95%CI 3.9 - 4.5), also more common in women - 5.2% (95%CI 4.7 - 5.6) - than in men - 3.0% (95%CI 2.7 - 3.4). The prevalence of angina by age group increased progressively with age. The prevalence of angina of any sort was inverse to years of formal study. Despite the higher value of the presence of angina in black people, there was no significant difference by race/skin color. Conclusion: The high prevalence rate of angina pectoris in the population aged 18 years and above was consistent with studies in other countries, revealing the importance of coronary heart disease as a public health problem.

"Omics" studies on carbon nanoparticles effects / Estudos "ômicos" sobre os efeitos das nanoparticulas de carbono

Nanoparticles are being explored for biomedical applications in diagnostics and therapeutics. However, biocompatibility, toxicity and cellular entry ability are critical factors that will determine the utility of nanoparticles in clinical applications. "Omics" technologies can provide powerful tools that are sensitive and complementary for the study of molecular toxicology and for biomedical applications of nanomaterials. Specifically, proteomics could elucidate mechanisms of action and contribute to studies on how nanomaterials affect protein expression in tissues and organs, metabolic pathways, signal transduction mechanisms and target molecules. The objective of this review is to present a small approach about "omics" technologies applied to the study of the effect of carbon nanomaterials in the cell.

As nanopartículas têm sido exploradas em aplicações biomédicas para fins diagnósticos e terapêuticos. No entanto, a biocompatibilidade, a toxicidade e a capacidade de entrada na célula são fatores críticos que irão determinar a utilidade de nanopartículas com aplicações clínicas. As chamadas tecnologias "ômicas" podem fornecer poderosas ferramentas sensíveis e complementares para o estudo de toxicologia molecular e para aplicações biomédicas dos nanomateriais. Especificamente a proteômica poderia elucidar mecanismos de ação e contribuir no estudo do efeito de um nanomaterial sobre a expressão de proteínas ao nível de tecidos e órgãos, alterações em vias metabólicas, mecanismos de transdução e de regulação de genes assim como os alvos moleculares da terapêutica. O objetivo desta revisão é apresentar uma breve abordagem sobre as tecnologias "ômicas" aplicadas ao estudo do efeito dos nanomateriais de carbono nas células.