Peptides for Coating TiO2 Implants: An In Silico Approach.

Titanium is usually used in the manufacturing of metal implants due to its biocompatibility and high resistance to corrosion. A structural and functional connection between the living bone and the surface of the implant, a process called osseointegration, is mandatory for avoiding prolonged healing, infections, and tissue loss. Therefore, osseointegration is crucial for the success of the implantation procedure. Osseointegration is a process mediated by bone-matrix progenitor cells' proteins, named integrins. In this study, we used an in silico approach to assemble and test peptides that can be strategically used in sensitizing TiO2 implants in order to improve osseointegration. To do so, we downloaded PDB structures of integrins α5ß1, αvß3, and αIIbß3; their biological ligands; and low-cost proteins from the Protein Data Bank, and then we performed a primary (integrin-protein) docking analysis. Furthermore, we modeled complex peptides with the potential to bind to the TiO2 surface on the implant, as well as integrins in the bone-matrix progenitor cells. Then we performed a secondary (integrin-peptide) docking analysis. The ten most promising integrin-peptide docking results were further verified by molecular dynamics (MD) simulations. We recognized 82 peptides with great potential to bind the integrins, and therefore to be used in coating TiO2 implants. Among them, peptides 1 (GHTHYHAVRTQTTGR), 3 (RKLPDATGR), and 8 (GHTHYHAVRTQTLKA) showed the highest binding stability during the MD simulations. This bioinformatics approach saves time and more effectively directs in vitro studies.

Titanium Dental Implants: An Overview of Applied Nanobiotechnology to Improve Biocompatibility and Prevent Infections.

This review addresses the different aspects of the use of titanium and its alloys in the production of dental implants, the most common causes of implant failures and the development of improved surfaces capable of stimulating osseointegration and guaranteeing the long-term success of dental implants. Titanium is the main material for the development of dental implants; despite this, different surface modifications are studied aiming to improve the osseointegration process. Nanoscale modifications and the bioactivation of surfaces with biological molecules can promote faster healing when compared to smooth surfaces. Recent studies have also pointed out that gradual changes in the implant, based on the microenvironment of insertion, are factors that may improve the integration of the implant with soft and bone tissues, preventing infections and osseointegration failures. In this context, the understanding that nanobiotechnological surface modifications in titanium dental implants improve the osseointegration process arouses interest in the development of new strategies, which is a highly relevant factor in the production of improved dental materials.

The Deposition of a Lectin from Oreochromis niloticus on the Surface of Titanium Dioxide Nanotubes Improved the Cell Adhesion, Proliferation, and Osteogenic Activity of Osteoblast-like Cells.

Titanium and its alloys are used as biomaterials for medical and dental applications, due to their mechanical and physical properties. Surface modifications of titanium with bioactive molecules can increase the osseointegration by improving the interface between the bone and implant. In this work, titanium dioxide nanotubes (TiO2NTs) were functionalized with a lectin from the plasma of the fish Oreochromis niloticus aiming to favor the adhesion and proliferation of osteoblast-like cells, improving its biocompatibility. The TiO2NTs were obtained by anodization of titanium and annealed at 400 °C for 3 h. The resulting TiO2NTs were characterized by high-resolution scanning electron microscopy. The successful incorporation of OniL on the surface of TiO2NTs, by spin coating, was demonstrated by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIE), and attenuated total reflection-Fourier transform infrared spectrum (ATR-FTIR). Our results showed that TiO2NTs were successfully synthesized in a regular and well-distributed way. The modification of TiO2NTs with OniL favored adhesion, proliferation, and the osteogenic activity of osteoblast-like cells, suggesting its use to improve the quality and biocompatibility of titanium-based biomaterials.

Effect of chitosan nanoparticles on the inhibition of Candida spp. biofilm on denture base surface.

OBJECTIVES: Chitosan nanoparticles (ChNPs) have antifungal effects, however there is a lack of information about the effects of ChNPs against Candida biofilm on denture base surface. This study investigated the ChNPs effect against C. albicans biofilm adhesion and formation, and against Candida spp. biofilm on heat-cured acrylic resin. DESIGN: The ChNPs were synthetized (3800 µg/mL) and characterized by infra-red spectrophotometry and transmission electron microscopy. The minimum inhibitory/fungicidal concentrations (MIC/MFC) against Candida spp. were determined. The time-kill assay and changes on C. albicans micromorphology were evaluated. The % inhibition of ChNPs on C. albicans biofilm formation and reduction were investigated using 1 min and 8 h exposure. Candida biofilm was developed on resin surfaces and ChNPs were applied every 8 h for 5 days. After, fungal cells were counted (CFU/mL) and the surface roughness (Ra) and vickers microhardness (HV) of resin were analysed. For all experiments, sodium hypochlorite (NaOCl) was used as control. Data were analyzed by ANOVA, Tukey and paired t-tests (α = 0.05). RESULTS: The MIC80% of ChNPs was 30.1 µg/mL. ChNPs at 4 MIC showed complete inhibition in the time-kill assays. Blastoconidia cells were predominant after ChNPs application. The % inhibition ChNPs on C. albicans was proportional to its concentration, regardless of the exposure time. ChNPs decreased the CFU/mL of Candida spp. and showed lower alteration of HV and Ra values of resin surface compared to NaOCl. CONCLUSIONS: The ChNPs inhibited C. albicans biofilm, reduced Candida biofilm on resin and caused small changes in roughness and hardness of acrylic resin surface.

Encapsulation of the HSP-90 Chaperone Inhibitor 17-AAG in Stable Liposome Allow Increasing the Therapeutic Index as Assessed, in vitro, on Leishmania (L) amazonensis Amastigotes-Hosted in Mouse CBA Macrophages.

The current long-term treatment for leishmaniasis causes severe side effects and resistance in some cases. An evaluation of the anti-leishmanial potential of an HSP90-inhibitor, 17-allylamino-17-demethoxygeldanamycin (17-AAG), demonstrated its potent effect against Leishmania spp. in vitro and in vivo. We have previously shown that 17-AAG can kill L. (L) amazonensis promastigotes with an IC50 of 65 nM and intracellular amastigote at concentrations as low as 125 nM. As this compound presents low solubility and high toxicity in human clinical trials, we prepared an inclusion complex containing hydroxypropyl-ß-cyclodextrin and 17-AAG (17-AAG:HPßCD) to improve its solubility. This complex was characterized by scanning electron microscopy, and X-ray diffraction. Liposomes-containing 17-AAG:HPßCD was prepared and evaluated for encapsulation efficiency (EE%), particle size, polydispersity index (PDI), pH, and zeta potential, before and after accelerated and long-term stability testing. An evaluation of leishmanicidal activity against promastigotes and intracellular amastigotes of L. (L) amazonensis was also performed. The characterization techniques utilized confirmed the formation of the inclusion complex, HPßCD:17-AAG, with a resulting 33-fold-enhancement in compound water solubility. Stability studies revealed that 17-AAG:HPßCD-loaded liposomes were smaller than 200 nm, with 99% EE. Stability testing detected no alterations in PDI that was 0.295, pH 7.63, and zeta potential +22.6, suggesting liposome stability, and suitability for evaluating leishmanicidal activity. Treatment of infected macrophages with 0.006 nM of 17-AAG:HPßCD or 17-AAG:HPßCD-loaded liposomes resulted in almost complete amastigote clearance inside macrophages after 48 h. This reduction is similar to the one observed in infected macrophages treated with 2 µM amphotericin B. Our results showed that nanotechnology and drug delivery systems could be used to increase the antileishmanial efficacy and potency of 17-AAG in vitro, while also resulting in reduced toxicity that indicates these formulations may represent a potential therapeutic strategy against leishmaniasis.

Evaluation of photodynamic activity, photostability and in vitro drug release of zinc phthalocyanine-loaded nanocapsules.

Nanocapsule formulations containing zinc phthalocyanine (ZnPc) were investigated as drug delivery systems for use in photodynamic therapy (PDT). ZnPc loaded chitosan, PCL, and PCL coated with chitosan nanocapsules were prepared and characterized by means of their physicochemical properties, photodynamic activity, photostability and drug release profile. All formulations presented nanometric hydrodynamic radius, around 100 nm, low polydispersity index (0.08-0.24), slightly negative zeta potential for PCL nanoparticles and positive zeta potential for suspension containing chitosan. Encapsulation efficiencies were higher than 99%. The capacity of ZnPc loaded nanocapsules to produce cytotoxic singlet oxygen ((1)O2) by irradiation with red laser was monitored using 1.3-diphenylisobenzofuran as a probe. The singlet oxygen quantum yields (ΦΔ) for ZnPc loaded chitosan nanocapsules were high and similar to that of the standard (ZnPc in DMSO), displaying excellent ability to generate (1)O2. The photosensitizer loaded nanocapsules are photostable in the timescale usually utilized in PDT and only a small photobleaching event was observed when a light dose of 610J/cm(2) was applied. The in vitro drug release studies of ZnPc from all nanocapsules demonstrated a sustained release profile controlled by diffusion, without burst effect. The nature of the polymer and the core type of the nanocapsules regulated ZnPc release. Thus, the nanocapsules developed in this work are a promising strategy to be employed in PDT.

Morphological and structural characterization of PHBV/organoclay nanocomposites by small angle X-ray scattering.

In this work, the morphological and structural behaviors of poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) nanocomposites were investigated using small angle X-ray scattering (SAXS), wide angle X-ray diffraction (WAXD) and transmission electron microscopy (TEM). The nanocomposites with 1, 3 and 5 wt.% of organically modified montmorillonite Cloisite® 30B (OMMT) were prepared by melt processing in a twin screw extruder using two different processing conditions (low and high shear intensity). The lamellar long period of the polymer was lower for the nanocomposites, with high polydispersity values. However, the crystalline thickness increased with the clay content and was independent of the processing conditions. This behavior resulted in a high linear crystallinity of the nanocomposites with 3 and 5 wt.% OMMT. The disruption factor (ß) was in agreement with the WAXD and TEM findings, indicating a good dispersion of the nanoparticles in the PHBV matrix with 3 wt.% of OMMT during the high shear intensity of melt processing.

Direct synthesis of coated gold nanoparticles mediated by polymers with amino groups.

The single-step/single-phase synthesis of hybrid organic-inorganic core-shell gold nanoparticles (AuNPs), facilitated by amino-functionalized amphiphilic block copolymers that simultaneously play the roles of reductant and stabilizer, was investigated in this study. Experiments were devised with emphasis on the pH-responsive poly(ethylene oxide)-b-poly(2,3-dihydroxypropyl methacrylate)-b-poly[2-(diisopropylamino)ethyl methacrylate] triblock copolymer, which allows direct chemical cross-linking of the micellar structures to be performed. The polymer structure-reactivity relationship associated with the AuNP formation was established using a set of six structurally related macromolecules. AuNP formation was dependent on the aqueous dissociation equilibrium involving tertiary amino groups, the Au(III) speciation, and electrochemical redox potentials. The effects of these parameters on the synthesis of AuNPs change as the solution pH is increased from pH 3.3 (molecularly dissolved polymer chains; no AuNP formation) to 6.8 or higher (polymer chains self-assembled into spherical micelles; stable gold sols are produced), and Au(III) reduction potentials shift toward the cathodic region while the oxidation potential of deprotonated amino groups decreases. Sigmoidal nanoparticle growth kinetics was observed in all cases after a characteristic induction period. Stable, well-defined, uniform polymer-coated gold colloids with localized surface plasmon resonance centered at 53 0nm can be conveniently produced in one-pot, two-reactant, no work-up reactions when the stoichiometry is [N]/[Au]=3.5-25.0.

Influence of different restorative techniques on marginal seal of class II composite restorations.

OBJECTIVE: To evaluate the gingival marginal seal in class II composite restorations using different restorative techniques. MATERIAL AND METHODS: Class II box cavities were prepared in both proximal faces of 32 sound human third molars with gingival margins located in either enamel or dentin/cementum. Restorations were performed as follows: G1 (control): composite, conventional light curing technique; G2: composite, soft-start technique; G3: amalgam/composite association (amalcomp); and G4: resin-modified glass ionomer cement/composite, open sandwich technique. The restored specimens were thermocycled. Epoxy resin replicas were made and coated for scanning electron microscopy examination. For microleakage evaluation, teeth were coated with nail polish and immersed in dye solution. Teeth were cut in 3 slices and dye penetration was recorded (mm), digitized and analyzed with Image Tool software. Microleakage data were analyzed statistically by non-parametric Kruskal-Wallis and Mann-Whitney tests. RESULTS: Leakage in enamel was lower than in dentin (p<0.001). G2 exhibited the lowest leakage values (p<0.05) in enamel margins, with no differences between the other groups. In dentin margins, groups G1 and G2 had similar behavior and both showed less leakage (p<0.05) than groups G3 and G4. SEM micrographs revealed different marginal adaptation patterns for the different techniques and for the different substrates. CONCLUSION: The soft-start technique showed no leakage in enamel margins and produced similar values to those of the conventional (control) technique for dentin margins.

Influence of different restorative techniques on marginal seal of class II composite restorations

OBJECTIVE: To evaluate the gingival marginal seal in class II composite restorations using different restorative techniques. MATERIAL AND METHODS: Class II box cavities were prepared in both proximal faces of 32 sound human third molars with gingival margins located in either enamel or dentin/cementum. Restorations were performed as follows: G1 (control): composite, conventional light curing technique; G2: composite, soft-start technique; G3: amalgam/composite association (amalcomp); and G4: resin-modified glass ionomer cement/composite, open sandwich technique. The restored specimens were thermocycled. Epoxy resin replicas were made and coated for scanning electron microscopy examination. For microleakage evaluation, teeth were coated with nail polish and immersed in dye solution. Teeth were cut in 3 slices and dye penetration was recorded (mm), digitized and analyzed with Image Tool software. Microleakage data were analyzed statistically by non-parametric Kruskal-Wallis and Mann-Whitney tests. RESULTS: Leakage in enamel was lower than in dentin (p<0.001). G2 exhibited the lowest leakage values (p<0.05) in enamel margins, with no differences between the other groups. In dentin margins, groups G1 and G2 had similar behavior and both showed less leakage (p<0.05) than groups G3 and G4. SEM micrographs revealed different marginal adaptation patterns for the different techniques and for the different substrates. CONCLUSION: The soft-start technique showed no leakage in enamel margins and produced similar values to those of the conventional (control) technique for dentin margins.