Proteome analysis of the plasma protein layer adsorbed to a rough titanium surface

In this study a label-free proteomic approach was used to investigate the composition of the layer of protein adsorbed to rough titanium (Ti) after exposure to human blood plasma. The influence of the protein layer on the surface free energy (SFE) of the Ti was evaluated by contact angle measurements. Ti discs were incubated with blood plasma for 180?min at 37?°C, and the proteins recovered were subjected to liquid chromatography coupled to tandem mass spectrometry analysis. A total of 129 different peptides were identified and assigned to 25 distinct plasma proteins. The most abundant proteins were fibronectin, serum albumin, apolipoprotein A-I, and fibrinogen, comprising 74.54% of the total spectral counts. Moreover, the protein layer increased the SFE of the Ti (p?<?0.05). The layer adsorbed to the rough Ti surface was composed mainly of proteins related to cell adhesion, molecule transportation, and coagulation processes, creating a polar and hydrophilic interface for subsequent interactions with host cells.     

Crystalline anatase-rich titanium can reduce adherence of oral streptococci

Dental implant abutments that emerge through the mucosa are rapidly covered with a salivary protein pellicle to which bacteria bind, initiating biofilm formation. In this study, adherence of early colonizing streptococci, Streptococcus gordonii, Streptococcus oralis, Streptococcus mitis and Streptococcus sanguinis to two saliva-coated anodically oxidized surfaces was compared with that on commercially pure titanium (CpTi). Near edge X-ray absorption (NEXAFS) showed crystalline anatase was more pronounced on the anodically oxidized surfaces than on the CpTi. As revealed by fluorescence microscopy, a four-species mixture, as well as individual bacterial species, exhibited lower adherence after 2?h to the saliva-coated, anatase-rich surfaces than to CpTi. Since wettability did not differ between the saliva-coated surfaces, differences in the concentration and/or configuration of salivary proteins on the anatase-rich surfaces may explain the reduced bacterial binding effect. Anatase-rich surfaces could thus contribute to reduced overall biofilm formation on dental implant abutments through diminished adherence of early colonizers.     

Proteomic profile of the saliva and plasma protein layer adsorbed on Ti–Zr alloy: the effect of sandblasted and acid-etched surface treatment

Abstract

Titanium–zirconium (Ti–Zr) alloy has been widely used as a biomaterial for implant devices, and it is commonly treated by sandblasting followed by acid etching (SLA) to improve biological responses. Although protein adsorption is the first biological response, the effect of this SLA treatment on the proteomic profile of proteins adsorbed from saliva and blood plasma has not been tested. In this study, the proteomic profile was evaluated by liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS). Streptococcus sanguinis was used to test whether the protein layer affects bacterial adhesion. SLA treatment affected the proteomic profile, showing exclusive proteins adsorbed from saliva (14) and plasma (3). However, both groups exhibited close patterns of intensity for common proteins, molecular functions and biological processes mediated by proteins. Interestingly, Ti–Zr_SLA showed higher bacterial adhesion (～1.9 fold over) for the surface coated with plasma proteins. Therefore, SLA treatment of Ti–Zr alloy changed the proteomic profile, which may affect bacterial adhesion.     

Proteomic Evaluation of Acquired Enamel Pellicle during In Vivo Formation

Acquired enamel pellicle (AEP) is a protein film that forms on the enamel surface of teeth by selective adsorption of proteins and peptides present in the mouth. This protein film forms the interface between enamel and the damage oral biofilm, which modulates the attachment of bacteria found in oral biofilm. The overall goal of this study was to gain insight into the biological formation of the human in vivo AEP. This study hypothesized that AEP is created by the formation of successive protein layers, which consist of initial binding to enamel and subsequent protein-protein interactions. This hypothesis was examined by observing quantitative and qualitative changes in pellicle composition during the first two hours of AEP formation in the oral cavity. Quantitative mass spectrometry approaches were used to generate an AEP protein profile for each time-point studied. Relative proteomic quantification was carried out for the 50 proteins observed in all four time-points. Notably, the abundance of important salivary proteins, such as histatin 1, decrease with increasing of the AEP formation, while other essential proteins such as statherin showed constant relative abundance in all time-points. In summary, this is the first study that investigates the dynamic process to the AEP formation by using proteomic approaches. Our data demonstrated that there are significant qualitative and quantitative proteome changes during the AEP formation, which in turn will likely impact the development of oral biofilms.     

Low Pressure Radio-Frequency Oxygen Plasma Induced Oxidation of Titanium – Surface Characteristics and Biological Effects

Objective

This research was designed to investigate the effects of low pressure radio-frequency (RF) oxygen plasma treatment (OPT) on the surface of commercially pure titanium (CP-Ti) and Ti6Al4V. Surface topography, elemental composition, water contact angle, cell viability, and cell morphology were surveyed to evaluate the biocompatibility of titanium samples with different lengths of OP treating time.

Materials and Methods

CP-Ti and Ti6Al4V discs were both classified into 4 groups: untreated, treated with OP generated by using oxygen (99.98%) for 5, 10, and 30 min, respectively. After OPT on CP-Ti and Ti6Al4V samples, scanning probe microscopy, X-ray photoelectron spectrometry (XPS), and contact angle tests were conducted to determine the surface topography, elemental composition and hydrophilicity, respectively. The change of surface morphology was further studied using sputtered titanium on silicon wafers. 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay and F-actin immunofluorescence stain were performed to investigate the viability and spreading behavior of cultivated MG-63 cells on the samples.

Results

The surface roughness was most prominent after 5 min OPT in both CP-Ti and Ti6Al4V, and the surface morphology of sputtered Ti sharpened after the 5 min treatment. From the XPS results, the intensity of Ti^°, Ti²⁺, and Ti³⁺ of the samples’ surface decreased indicating the oxidation of titanium after OPT. The water contact angles of both CP-Ti and Ti6Al4V were increased after 5 min OPT. The results of MTT assay demonstrated MG-63 cells proliferated best on the 5 min OP treated titanium sample. The F-actin immunofluorescence stain revealed the cultivated cell number of 5 min treated CP-Ti/Ti6Al4V was greater than other groups and most of the cultivated cells were spindle-shaped.

Conclusions

Low pressure RF oxygen plasma modified both the composition and the morphology of titanium samples’ surface. The CP-Ti/Ti6Al4V treated with 5 min OPT displayed the roughest surface, sharpest surface profile and best biocompatibility.     

Comparative analysis of corrosion resistance between beta titanium and Ti-6Al-4V alloys: A systematic review

BackgroundThe knowledge of the electrochemical property (corrosion resistance) of beta titanium alloys compared to Ti-6Al-4 V for implants is relevant because of the potential cytotoxic effects that the released ions could cause to long-term health.ObjectivesThe objective of this systematic review was to seek information on the electrochemical properties (corrosion resistance) of beta titanium alloys compared to Ti-6Al-4 V since the awareness of the electrochemical behavior of the implant surface in the medium is essential for the best indication of the alloys or compositional changes, which may promote benefits to bone-implant interaction in all areas that this procedure is required.MethodsThe PubMed, LILACS, COCHRANE Library, and Science Direct databases were electronically searched for the terms: dental implants AND beta-titanium AND Ti-6Al-4 V AND electrochemical technics. The inclusion criteria were research articles that studied beta-titanium compared to Ti-6Al-4 V using electrochemical techniques in electrolytes of chemical composition similar to body fluid, published in English, between 2000 and 2020. Articles that did not compare the corrosion resistance of these alloys in electrolytes similar to body fluids were excluded.ResultsA total of 189 articles were restored and selected by title and/or abstract according to the inclusion and exclusion criteria, which resulted in 15 articles that were reduced to 8 after read in full. The studies in vitro evaluated the corrosion resistance in electrolytes Hank, Ringer, SBF, and 0.9 % NaCl, between beta titanium alloys, obtained by arc fusion or bars stock, and Ti-6Al-4 V, for dental or biomedical implants submitted to surface treatments by heat treatment, plasma electrolytic oxidation (PEO), alkaline treatment, and thermomechanical.ConclusionThe evaluated literature allowed to determine that 1) The oxides Nb₂O ₅, Ta₂O _5, and ZrO₂ have higher stability and protection quality than that of TiO₂ modified by the oxides of Al and V; 2) A higher modulus of elasticity of the Ti-6Al-4 V alloy favors protection against corrosion by maintaining a thicker and more firmly adhered oxide layer; 3) The increase in the thickness of the Ti alloys superficial layer contributes to the improvement of the corrosion resistance.     

The composition of enamel salivary films is different from the ones formed on dental materials

This study utilized two-dimensional gel electrophoresis (2DE) to illustrate the compositional differences between in vitro salivary conditioning films (denoted pellicles) formed on human enamel as well as on the dental materials titanium and poly(methyl methacrylate). The salivary pellicles were formed by immersing each surface in individual tubes containing small volumes of freshly collected whole saliva. Saliva remaining in the tubes after the pellicle formation for 2 h was visualized by means of 2DE and silver staining. The results showed that the protein patterns in 2DE of the liquid phase of saliva left after the exposure to the respective surfaces, regarding proteins <100 kDa in size, were different depending on the surface used. Several protein groups and/or individual proteins were shown to be distinct for each surface used.     

Validation of mechanically-assisted sodium dodecyl-sulphate elution as a technique to remove pellicle protein components from human enamel

The salivary film, denoted the pellicle, formed on oral surfaces is of great importance for oral health and comfort. The present study describes mechanically-assisted sodium dodecyl sulphate (SDS) elution of the in vivo pellicle formed on human enamel and visualisation of the desorbed pellicle proteins using two-dimensional gel electrophoresis (2-DE). To verify this removal of the pellicle, a combined mechanical and surfactant procedure was additionally performed on an in vitro pellicle formed on human enamel, and the effectiveness was validated by mechanical removal in combination with HCl. As indicated by protein quantitation and one dimensional gel electrophoresis, rubbing with polyamide fibre pellets soaked in a 0.5% SDS solution was optimal for completely removing the adsorbed proteins from the enamel surface, and yet provided separation of the proteins by 2-DE to enable identification in future studies.     

Biomaterials for orthopedics: anti-biofilm activity of a new bioactive glass coating on titanium implants

Abstract

This study evaluated adhesion and biofilm formation by Candida albicans, Pseudomonas aeruginosa and Staphylococcus epidermidis on surfaces of titanium (Ti) and titanium coated with F18 Bioactive Glass (BGF18). Biofilms were grown and the areas coated with biofilm were determined after 2, 4 and 8?h. Microscopy techniques were applied in order to visualize the structure of the mature biofilm and the extracellular matrix. On the BGF18 specimens, there was less biofilm formation by C. albicans and S. epidermidis after incubation for 8?h. For P. aeruginosa biofilm, a reduction was observed after incubation for 4?h, and it remained reduced after 8?h on BGF18 specimens. All biofilm matrices seemed to be thicker on BGF18 surface than on titanium surfaces. BGF18 showed significant anti-biofilm activity in comparison with Ti in the initial periods of biofilm formation; however, there was extensive biofilm after incubation for 48?h.     

Photocatalytic antibacterial effects on TiO2-anatase upon UV-A and UV-A/VIS threshold irradiation

Photocatalysis mediated by the anatase modification of titanium dioxide (TiO₂) has shown antibacterial effects in medical applications. The aim of this study was to investigate the possibility of expanding the excitation wavelengths for photocatalytic antibacterial effects from ultraviolet (UV) into the visible light range. After deposition of salivary pellicle and adhesion of Streptococcus gordonii on anatase, different irradiation protocols were applied to induce photocatalysis: ultraviolet A (UV-A) > 320 nm; ultraviolet/visible (UV-A/VIS) light > 380 nm and > 390 nm; and VIS light 400–410 nm. A quartz crystal microbalance with dissipation (QCM-D) tests and microscopic examination were used to observe the photoinduced antibacterial effects. Salivary pellicle could be photocatalytically decomposed under all irradiation protocols. In contrast, effective photocatalytic attack of bacteria could be observed by UV-A as well as by UV-A/VIS at 380 nm < λ < 390 nm only. Wavelengths above 380 nm show promise for in situ therapeutic antifouling applications.     

Biomechanical assessment of a PEEK rod system for semi-rigid fixation of lumbar fusion constructs

The concept of semi-rigid fixation (SRF) has driven the development of spinal implants that utilize nonmetallic materials and novel rod geometries in an effort to promote fusion via a balance of stability, intra- and inter-level load sharing, and durability. The purpose of this study was to characterize the mechanical and biomechanical properties of a pedicle screw-based polyetheretherketone (PEEK) SRF system for the lumbar spine to compare its kinematic, structural, and durability performance profile against that of traditional lumbar fusion systems. Performance of the SRF system was characterized using a validated spectrum of experimental, computational, and in vitro testing. Finite element models were first used to optimize the size and shape of the polymeric rods and bound their performance parameters. Subsequently, benchtop tests determined the static and dynamic performance threshold of PEEK rods in relevant loading modes (flexion-extension (F/E), axial rotation (AR), and lateral bending (LB)). Numerical analyses evaluated the amount of anteroposterior column load sharing provided by both metallic and PEEK rods. Finally, a cadaveric spine simulator was used to determine the level of stability that PEEK rods provide. Under physiological loading conditions, a 6.35 mm nominal diameter oval PEEK rod construct unloads the bone-screw interface and increases anterior column load (approx. 75% anterior, 25% posterior) when compared to titanium (Ti) rod constructs. The PEEK construct's stiffness demonstrated a value lower than that of all the metallic rod systems, regardless of diameter or metallic composition (78%??80% reduction in F/E, p??70% reduction in LB, p??54% reduction in AR, p?相似文献   

Pellicle structure in Euglena

The pellicle of Euglena has been investigated by anoptral and phase contrast light microscopy, and by electron microscopy of osmium-fixed, Epon-embedded, lead-stained sections and of carbon/platinum replicas.

Observations on living cells show that the pellicular striatiors of Euglena spirogyra trace a left-handed (S) helix in a majority of cells, and a right-handed (Z) helix in only 5 to 30% of the cells in any one culture. All cells of E. spirogyra var. fusca have a left-handed (S) helix. Ornamentation on the pellicle takes the form of rows of knobs in various patterns. The living pellicle can be dissociated into long flat strips.

Electron microscopy shows that each pellicular strip has an elaborate cross-sectional shape, features of which are accessory teeth and ribs and a continuous ridge which articulates in a groove running along the edge of the next strip. The strips can move against one another, presumably by the ridges sliding in the grooves, and it is suggested that the joints might be lubricated by mucilage supplied from the helically disposed muciferous bodies. One single and one pair of fibrils or tubes, 200–250 Å in diameter, are regularly arranged parallel to each pellicular strip. A continuous tripartite plasmalemma, 80–100 Å thick, lies externally to the strips; external to this membrane lie the pellicular knobs. Each cell has from 35 to 45 pellicular strips, reducing to a few at the posterior end of the cell by successive fusions. Similar fusions occur at the anterior end of the cell, mainly within the canal.

These observations are compared with those made on the euglenoid pellicle by previous authors, and the following problems are discussed: direction of helix; the nature and cause of ornamentation; euglenoid movement with reference to fibrils, cytoplasmic flow, pellicle flexibility and the proteinaceous nature of the pellicle; helical and bilateral symmetry in the cell; and cet growth and division.     

Reduction of bacterial adhesion on titanium-doped diamond-like carbon coatings

A range of titanium doped diamond-like carbon (Ti-DLC) coatings with different Ti contents were prepared on stainless steel substrates using a plasma-enhanced chemical vapour deposition technique. It was found that both the electron donor surface energy and the surface roughness of the Ti-DLC coatings increased with increasing Ti contents in the coatings. Bacterial adhesion to the coatings was evaluated against Escherichia coli WT F1693 and Pseudomonas aeruginosa ATCC 33347. The experimental data showed that bacterial adhesion decreased with the increases of the Ti content, the electron donor surface energy and surface roughness of the coatings, while the bacterial removal percentage increased with the increases of these parameters. The Ti-DLC coatings reduced bacterial attachment by up to 75% and increased bacterial detachment from 15 to 45%, compared with stainless steel control.     

Interactions between cells and titanium surfaces

The interaction between cells and implant materials is determined by the surface structure and/or surface composition of the material. In the past years, titanium and titanium alloys have proved their superiority over other implant materials in many clinical applications. This predominant behaviour is caused by a dense passive oxide layer which forms within milliseconds in oxidizing media. Titanium dioxide layers of 100 nm thickness were produced on the surface of cp-titanium grade 2, and on an experimental alloy of high vanadium content (Ti1.5Al25V) as a harmful control. The layers were produced by thermal and anodic oxidation and by coating by means of the sol-gel process. The resulting oxide layers were characterized with respect of their structure and chemical composition. In cell tests (proliferation, MTT, morphology, actin staining), the reaction of the cells was examined. It was shown that the sol-gel-produced titanium oxide layer is able to shield the cells from toxic alloying elements, with the result that the cell reaction is influenced only by the thin titanium oxide surface layer and not by the composition of the bulk material.     

Increased transglutaminase activity was associated with IL-6 release in cultured human gingival fibroblasts exposed to dental cast alloys

Summary. Molecular mechanisms underlying gingival and periodontal inflammation caused by dental alloys are still poorly understood. Recently, it has been demonstrated that tissue transglutaminase can be involved in inflammatory cell response. The aim of this study was to evaluate effects of exposure to orthodontic materials on transglutaminase in cultured human gingival fibroblasts. The incubation with Ni–Ti heat-activated (T3) or Ni–Ti super-elastic (T4), and with Ni–Cr–Co (T2) alloys produced respectively 2.5-fold and 8-fold increases in IL-6 release compared with control cultures. Transglutaminase activity was significantly increased in cells exposed to T3 and T4 alloys (about 170% of control; p < 0.05), where it was mainly localized close to inner part of cell membrane. The exposure to T3 and T4 specimens significantly up-regulated also tTG expression compared with control cultures. These data first show an association between IL-6 release and tissue transglutaminase increases, suggesting that TGase-mediated reactions may play a major role in periodontal inflammation.     

Influence of Bio-Lubricants on the Tribological Properties of Ti6Al4V Alloy

Titanium alloy is one of the best materials for biomedical applications due to its superior biocompatibility, outstanding corrosion resistance, and low elastic modulus. However, the friction and wear behaviors of titanium alloys were sensitive to the environment including lubrication. In order to clarify the wear mechanism of titanium alloy under different lubrications including deionized water, physiological saline and bovine serum, the friction and wear tests were performed between Ti6Al4V plates and Si₃N₄ ball on a universal multi-functional tester. The friction and the wear rate of titanium alloy were measured under dry friction and three different lubrication conditions. The worn surfaces were examined by scanning electron microscopy. The results revealed that under the dry friction, the wear resistance of titanium alloy was the worst since the wear mechanism was mainly the combination of abrasive wear and oxidation wear. It was also found that Ti6Al4V alloy had low friction coefficient and wear rate under three lubrication conditions, and its wear mechanism was adhesive wear.     

Morphology and chemical characterization of Ti surfaces modified for biomedical applications

The aim of the present work is to characterize in detail the chemical composition and morphology of titanium surfaces subjected to various environments. Modifications consisted of exposure of Ti to acidic, alkaline or polymer solutions. Such modifications result in chemical and/or morphological changes in the Ti surface. Special attention has been given to identifying the factors influencing cell adhesion and growth. SEM examinations provided morphological characterization of the Ti samples. Surface analytical techniques such as AES or XPS combined with Ar(+) ion sputtering allowed examination of the chemical properties of the Ti surface after chemical pretreatments and investigating the chemical composition of the Ti oxide layer. Raman spectroscopy investigations allowed determination of the crystalline phases of the Ti-oxide layers and characterization of the dextran-modified surface. The results show large differences in the morphology of Ti pretreated with different procedures whereas only minor differences in the chemistry of the surfaces were found. High-resolution Auger investigations have revealed that all the chemical modifications of Ti surfaces resulted in the formation of a titanium oxide layer. XPS confirmed that TiO(2) is the main component of the chemically modified Ti surface. The Raman spectroscopy investigations showed that the titanium surface with a dextran coating is rich in hydroxyl groups. All the surfaces investigated exhibit a hydrophilic character. The possible influence of various surface features on surface biocompatibility is discussed.     

Quantitative proteomic analysis of the fall armyworm saliva

Lepidopteran larvae secrete saliva on plant tissues during feeding. Components in the saliva may aid in food digestion, whereas other components are recognized by plants as cues to elicit defense responses. Despite the ecological and economical importance of these plant-feeding insects, knowledge of their saliva composition is limited to a few species. In this study, we identified the salivary proteins of larvae of the fall armyworm (FAW), Spodoptera frugiperda; determined qualitative and quantitative differences in the salivary proteome of the two host races—corn and rice strains—of this insect; and identified changes in total protein concentration and relative protein abundance in the saliva of FAW larvae associated with different host plants. Quantitative proteomic analyses were performed using labeling with isobaric tags for relative and absolute quantification followed by liquid chromatography-tandem mass spectrometry. In total, 98 proteins were identified (>99% confidence) in the FAW saliva. These proteins were further categorized into five functional groups: proteins potentially involved in (1) plant defense regulation, (2) herbivore offense, (3) insect immunity, (4) detoxification, (5) digestion, and (6) other functions. Moreover, there were differences in the salivary proteome between the FAW strains that were identified by label-free proteomic analyses. Thirteen differentially identified proteins were present in each strain. There were also differences in the relative abundance of eleven salivary proteins between the two FAW host strains as well as differences within each strain associated with different diets. The total salivary protein concentration was also different for the two strains reared on different host plants. Based on these results, we conclude that the FAW saliva contains a complex mixture of proteins involved in different functions that are specific for each strain and its composition can change plastically in response to diet type.     

Microstructure analysis and wear behavior of titanium cermet femoral head with hard TiC layer

Titanium cermet was successfully synthesized and formed a thin gradient titanium carbide coating on the surface of Ti6Al4V alloy by using a novel sequential carburization under high temperature, while the titanium cermet femoral head was produced. The titanium cermet phase and surface topography were characterized with X-ray diffraction (XRD) and backscattered electron imaging (BSE). And then the wear behavior of titanium cermet femoral head was investigated by using CUMT II artificial joint hip simulator. The surface characterization indicates that carbon effectively diffused into the titanium alloys and formed a hard TiC layer on the Ti6Al4V alloys surface with a micro-porous structure. The artificial hip joint experimental results show that titanium cermet femoral head could not only improve the wear resistance of artificial femoral head, but also decrease the wear of UHMWPE joint cup. In addition, the carburized titanium alloy femoral head could effectively control the UHMWPE debris distribution, and increase the size of UHMWPE debris. All of the results suggest that titanium cermet is a prospective femoral head material in artificial joint.     

Burning mouth syndrome (BMS): sialometric and sialochemical analysis and salivary protein profile

Objective: The aim of the present study was to analyse the characteristics of salivary production and its composition in individuals with burning mouth syndrome (BMS). Study Design: Salivary flow rate, concentrations of potassium, iron, chloride, thiocyanate, magnesium, calcium, phosphorus, glucose, total protein and urea, as well as the expression profile of salivary proteins were analysed by SDS‐PAGE. Results: The mean salivary flow rate among control patients was lower than that of BMS patients. Chloride, phosphorus and potassium levels were elevated in patients with BMS (p = 0.041, 0.001 and 0.034, respectively). Total salivary protein concentration was reduced in individuals with BMS (p = 0.223). Analysis of the expression of salivary proteins by Coomassie blue SDS‐PAGE revealed a lower expression of low molecular weight proteins in individuals with BMS compared to healthy controls. Conclusions: These results indicate that the identification and characterisation of low molecular weight salivary proteins in BMS may be important in understanding BMS pathogenesis, thus contributing to its diagnosis and treatment.