Surface Properties Of Titanium Research Articles

Treatment considerations in peri-implantitis

Peri-implantitis is defined as an inflammatory process affecting the tissues around an osseointegrated implant, resulting in the loss of the supporting bone. Microbial adherence and colonization appear to play a major role in the pathogenesis of periimplantitis. The decision regarding treatment strategies is based on the diagnosis. The severity of the peri-implant lesion and the treatment strategies must include mechanical cleaning (infection control) procedures. Mechanical instrumentation is widely used for the debridement of dental implants, but this may alter the titanium's surface properties. Therefore, selection of the type of instrumentation should be made depending on the type of surface to be debrided. Also, patients with dental implants must always be enrolled in a supportive therapy program.

Effects of storage medium and UV photofunctionalization on time-related changes of titanium surface characteristics and biocompatibility.

Storage in aqueous solution and ultraviolet (UV) photofunctionalization are two applicable methods to overcome the biological aging and increase the bioactivity of titanium. As information regarding the combined effects of storage medium and UV photofunctionalization has never been found in published literatures, this study focused on whether appropriate storage methods and UV photofunctionalization have synergistic effects on the biological properties of aged titanium surfaces. Titanium plates and discs were sandblasted and acid etched and then further prepared in five different modes as using different storage mediums (air or dH2 O) for 4 weeks and then with or without UV treatment. The surface characteristics were evaluated with scanning electron microscopy, contact angle measurements, and X-ray photoelectron spectroscopy. MC3T3-E1 cells were cultured on the surfaces, and cellular morphology, proliferation, alkaline phosphatase activity, and osteocalcin release were evaluated. The results showed that nanostructures were observed on water-stored titanium surfaces with a size of about 15 × 20 nm(2) . UV treatment was effective to remove the hydrocarbon contamination on titanium surfaces stored in either air or water. UV photofunctionalization further enhanced the already increased bioactivity of modSLA on initial cell attachment, proliferation, alkaline phosphatase activity, and osteocalcin release. Overall, UV photofunctionalization was effective in further enhancing the already increased bioactivity by using dH2 O as storage medium, and the effect of UV treatment was much more overwhelming than that of the storage medium. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 932-940, 2016.

Differentiation, apoptosis, and GM-CSF receptor expression of human gingival fibroblasts on a titanium surface treated by a dual acid-etched procedure.

Analysis of the effects of titanium surface properties on the biological behavior of human gingival fibroblasts (HGFs). HGFs were in vitro cultured on a titanium surface modified by a dual acid-etched procedure and on a control machined surface. Cell adhesion, proliferation, apoptosis, production of certain extracellular matrix (ECM) proteins, and expression of granulocyte macrophage-colony stimulating factor receptor (GM-CSFR) were investigated using in each experiment a total of 18 samples for each titanium surface. Cell attachment at 3 h of culture was statistically significantly higher on the etched surface. HGF growth increased on both surfaces during the entire experimental period and at day 14 of culture cell proliferation was statistically significantly higher on the treated surface than on the control. No statistically significant differences in percentage of apoptosis events were observed between the surfaces. ECM protein production increased progressively over time on both surfaces. A statistically significant deposition was observed at day 7 and 14 for collagen I and only at day 14 for fibronectin and tenascin, when compared to the baseline. GM-CSFR registered a positive expression on both surfaces, statistically significant at day 14 on the etched surface in comparison with the machined one. Data showed that titanium surface microtopography modulates in vitro cell response and phenotypical expression of HGFs. The etched surface promoted a higher cell proliferation and differentiation improving the biological behavior of HGFs. Results suggest a possible beneficial effect of surface etching modification on peri-implant biological integration and soft tissue healing which is critical for the formation of a biological seal around the neck of dental implants.

Wettability studies of topologically distinct titanium surfaces

Biomedical implants made of titanium-based materials are expected to have certain essential features including high bone-to-implant contact and optimum osteointegration, which are often influenced by the surface topography and physicochemical properties of titanium surfaces. The surface structure in the nanoscale regime is presumed to alter/facilitate the protein binding, cell adhesion and proliferation, thereby reducing post-operative complications with increased lifespan of biomedical implants. The novelty of our TiO2 nanostructures lies mainly in the high level control over their morphology and roughness by mere compositional change and optimisation of the experimental parameters. The present work focuses on the wetting behaviour of various nanostructured titanium surfaces towards water. Kinetics of contact area of water droplet on macroscopically flat, nanoporous and nanotubular titanium surface topologies was monitored under similar evaporation conditions. The contact area of the water droplet on hydrophobic titanium planar surface (foil) was found to decrease during evaporation, whereas the contact area of the droplet on hydrophobic nanorough titanium surfaces practically remained unaffected until the complete evaporation. This demonstrates that the surface morphology and roughness at the nanoscale level substantially affect the titanium dioxide surface–water droplet interaction, opposing to previous observations for microscale structured surfaces. The difference in surface topographic nanofeatures of nanostructured titanium surfaces could be correlated not only with the time-dependency of the contact area, but also with time-dependency of the contact angle and electrochemical properties of these surfaces.

Titanium Surface Roughing Treatments contribute to Higher Interaction with Salivary Proteins MG2 and Lactoferrin.

Some surface treatments performed on titanium can alter the composition of salivary pellicle formed on this abiotic surface. Such treatments modify the titanium's surface properties and can promote higher adsorption of proteins, which allow better integration of titanium to the biotic system. This study aimed to evaluate the interactions between salivary proteins and titanium disks with different surface treatments. Machined titanium disks (n = 48) were divided into four experimental groups (n = 12), according to their surface treatments: surface polishing (SP); acid etching (A); spot-blasting plus acid etching (SB-A); spot-blasting followed by acid etching and nano-functionalization (SB-A-NF). Titanium surfaces were characterized by surface roughness and scanning electron microscopy (SEM). Specimens were incubated with human saliva extracted from submandibular and sublingual glands. Total salivary protein adsorbed to titanium was quantified and samples were submitted to western blotting for mucin glycoprotein 2 (MG2) and lactoferrin identification. Surface roughness was statistically higher for SB-A and SB-A-NF groups. Scanning electron microscopy images confirmed that titanium surface treatments increased surface roughness with higher number of porous and scratches for SB-A and SB-A-NF groups. Total protein adsorption was significantly higher for SB-A and SB-A-NF groups (p < 0.05), which also presented higher interactions with MG2 and lactoferrin proteins. The roughing of titanium surface by spot-blasting plus acid etching treatments contribute to higher interaction with salivary proteins, such as MG2 and lactoferrin. Titanium surface roughing increases the interactions of the substratum with salivary proteins, which can influence the integration of dental implants and their components to the oral environment. However, those treatments should be used carefully intraorally, avoiding increase biofilm formation.

Self-protective Oxide Nano-Coatings for Enhanced Surface Biocompatibility of Titanium

ABSTRACTThe biocompatibility of an implant material depends on the bulk physical properties in addition to the surface properties. In biomedical engineering and industry Ti and Ti-alloys are very popular biological implant material for their bulk physical properties and strength to weight ratio resembling those of nature bone. It is possible to modify the surface properties of titanium for enhanced surface biocompatibility. The main objective of the this study is to engineer a smart Ti-based prosthesis surface by self induced chemically modified titanium oxide nano-film by the chemical mechanical polishing process (CMP). This new process applied on bio-implants aims at significantly reducing the out-diffusion of Ti and other metallic impurities from prosthesis in contact with body fluids and tissue and simultaneously enhancing the surface mechanical, chemical and biological properties. CMP technique enables the growth of a thicker and denser self-protective native oxide on Ti and Ti alloy samples, while simultaneously inducing a controlled surface roughness. It is demonstrated that the Ti based dental implants with self-protective oxide induced surfaces help minimize chemical and bacterial reactivity in addition to Ti ion dissolution while promoting their biocompatibility through surface patterning. The studied self-protective oxide films can also be utilized for many additional applications including bio-sensors.

Bioactive Titanium Surfaces with the Effect of Inhibiting Biofilm Formation

Titanium metals and its alloy have been widely used in hard tissue repairing fields due to their good biocampatibility and mechanical properties. However, bioinert response and biomaterial associated infections are the main problems for their clinical application. In this study, we chose titanium plates treated with anodic oxidation (AO-Ti), alkali-heat (AH-Ti) and acid-alkali (AA-Ti) methods, which have been proved to be bioactive in vivo, to culture with Staphylococcus aureus and Escherichia coli to investigate the interaction between bioactive titanium surfaces and biofilm. We used X-ray diffraction (XRD), Scanning Electron Microscope (SEM), roughness measurement to study the physical-chemical properties of the as-received bioactive titanium surfaces, and Confocal Laser Scanning Microscope (CLSM) was employed to study the properties of biofilm formed on the biomaterial surfaces. The results indicate that the titanium surface subjected to anodic oxidation treatment is unfavorable for the formation of biofilm in vitro because the titania (TiO2) coating formed by anodizing has superior antimicrobial property than the other surfaces. Therefore, anodic oxidation surface modification is effective to endow titanium surface with bioactivity and antimicrobial property, which has the potential to improve the successful rate of the clinical application of titanium implants.

Osteoblastic response to pectin nanocoating on titanium surfaces

Osseointegration of titanium implants can be improved by organic and inorganic nanocoating of the surface. The aim of our study was to evaluate the effect of organic nanocoating of titanium surface with unmodified and modified pectin Rhamnogalacturonan-Is (RG-Is) isolated from potato and apple with respect to surface properties and osteogenic response in osteoblastic cells.Nanocoatings on titanium surfaces were evaluated by scanning electron microscopy, contact angle measurements, atomic force microscopy, and X-ray photoelectron spectroscopy. The effect of coated RG-Is on cell adhesion, cell viability, bone matrix formation and mineralization was tested using SaOS-2 cells. Nanocoating with pectin RG-Is affected surface properties and in consequence changed the environment for cellular response. The cells cultured on surfaces coated with RG-Is from potato with high content of linear 1.4-linked galactose produced higher level of mineralized matrix compared with control surfaces and surfaces coated with RG-I with low content of linear 1.4-linked galactose. The study showed that the pectin RG-Is nanocoating not only changed chemical and physical titanium surface properties, but also specific coating with RG-Is containing high amount of galactan increased mineralized matrix formation of osteoblastic cells in vitro.

The angiogenic behaviors of human umbilical vein endothelial cells (HUVEC) in co-culture with osteoblast-like cells (MG-63) on different titanium surfaces

ObjectivesInteraction between osteogenesis and angiogenesis plays an important role in implant osseointegration. In the present study we investigated the influence of titanium surface properties on the angiogenic behaviors of endothelial cells grown in direct contact co-culture with osteoblasts. MethodsHuman umbilical vein endothelial cells (HUVECs) and osteoblast-like cells (MG-63 cells) were grown in direct co-culture on the following titanium surfaces: acid-etched (A), hydrophilic A (modA), coarse-gritblasted and acid-etched (SLA) and hydrophilic SLA (SLActive). Cell proliferation was evaluated by cell counting combined with flow cytometry. The expression of von Willebrand Factor (vWF), thrombomodulin (TM), endothelial cell protein C receptor (EPCR), E-Selectin, as well as vascular endothelial growth factor (VEGF) receptors Flt-1 and KDR in HUVECs and VEGF in MG-63 were measured by qPCR. The dynamic behavior of endothelial cells was recorded by time-lapse microscopy. ResultsProliferation of HUVECs was highest on A, followed by SLA, modA and SLActive surfaces. The expression of vWF, TM, EPCR, E-Selectin and Flt-1 in HUVECs was significantly higher on A than on all other surfaces. The expression of KDR in HUVECs grown on A surface was below detection limit. VEGF expression in MG-63 cells was significantly higher on SLActive vs SLA and modA vs A surfaces. Time-lapse microscopy revealed that HUVECs moved quickest and formed cell clusters earlier on A surface, followed by SLA, modA and SLActive surface. ConclusionsIn co-culture conditions, proliferation and expression of angiogenesis associated genes in HUVECs are promoted by smooth hydrophobic Ti surface, which is in contrast to previous mono-culture studies.

Microstructure and Properties of Laser Surface Carburized Titanium and Titanium Alloys

Titanium and titanium alloys are widely used in many fields due to their some characteristics such as light density, high strength and excellent corrosion resistance. However, their practical applications are limited by their poor surface mechanical properties. Laser surface carburizing can improve the surface properties of titanium and titanium via the formation of titanium carbide and other reinforcing phases. In this paper, the properties of the alloyed coatings, the growth morphologies and the formation mechanism of titanium carbide are discussed.

Silver deposition on titanium surface by electrochemical anodizing process reduces bacterial adhesion of Streptococcus sanguinis and Lactobacillus salivarius.

The aim of this study was to determine the antibacterial properties of silver-doped titanium surfaces prepared with a novel electrochemical anodizing process. Titanium samples were anodized with a pulsed process in a solution of silver nitrate and sodium thiosulphate at room temperature with stirring. Samples were processed with different electrolyte concentrations and treatment cycles to improve silver deposition. Physicochemical properties were determined by X-ray photoelectron spectroscopy, contact angle measurements, white-light interferometry, and scanning electron microscopy. Cellular cytotoxicity in human fibroblasts was studied with lactate dehydrogenase assays. The in vitro effect of treated surfaces on two oral bacteria strains (Streptococcus sanguinis and Lactobacillus salivarius) was studied with viable bacterial adhesion measurements and growth curve assays. Nonparametric statistical Kruskal-Wallis and Mann-Whitney U-tests were used for multiple and paired comparisons, respectively. Post hoc Spearman's correlation tests were calculated to check the dependence between bacteria adhesion and surface properties. X-ray photoelectron spectroscopy results confirmed the presence of silver on treated samples and showed that treatments with higher silver nitrate concentration and more cycles increased the silver deposition on titanium surface. No negative effects in fibroblast cell viability were detected and a significant reduction on bacterial adhesion in vitro was achieved in silver-treated samples compared with control titanium. Silver deposition on titanium with a novel electrochemical anodizing process produced surfaces with significant antibacterial properties in vitro without negative effects on cell viability.

Controlled oxidative nanopatterning of microrough titanium surfaces for improving osteogenic activity.

To further enhance the biological properties of acid-etched microrough titanium surfaces, titania nanotextured thin films were produced by simple chemical oxidation, without significantly altering the existing topographical and roughness features. The nanotextured layers on titanium surfaces can be controllably varied by tuning the oxidation duration time. The oxidation treatment significantly reduced water contact angles and increased the surface energy compared to the surfaces prior to oxidation. The murine bone marrow stromal cells (BMSCs) were used to evaluate the bioactivity. In comparison, oxidative nanopatterning of microrough titanium surfaces led to improved attachment and proliferation of BMSCs. The rate of osteoblastic differentiation was also represented by the increased levels of alkaline phosphatase activity and mineral deposition. These data indicated that oxidative nanopatterning enhanced the biological properties of the microrough titanium surfaces by modulating their surface chemistry and nanotopography. Based on the proven mechanical interlocking ability of microtopographies, enhancement of multiple osteoblast functions attained by this oxidative nanopatterning is expected to lead to better implant osseointegration in vivo.

Instrumentation With Ultrasonic Scalers Facilitates Cleaning of the Sandblasted and Acid-Etched Titanium Implants

Mechanical instrumentation is widely used to debride dental implants, but this may alter the surface properties of titanium, which in turn may influence bacterial adhesion and make it more difficult to remove the biofilm. This in vitro study was performed (1) to assess the amount of biofilm formation on a sand-blasted and acid-etched titanium fixture treated with ultrasonic scalers with metal, plastic, and carbon tips and (2) to evaluate how this treatment of titanium surfaces affects implant cleaning by brushing with dentifrice. The titanium fixtures were treated with various ultrasonic scaler tips, and surface roughness parameters were measured by confocal microscopy. Biofilm was formed on the treated fixtures by using pooled saliva from 10 subjects, and the quantity of the adherent bacteria was compared with crystal violet assay. The fixture surfaces with biofilm were brushed for total of 30 seconds with a toothbrush with dentifrice. The bacteria remaining on the brushed fixture surfaces were quantified by scanning electron microscopy. Surface changes were evident, and the changes of the surfaces were more discernible when metal tips were used. A statistically significant decrease in roughness value (arithmetic mean height of the surface) was seen in the 2 metal-tip groups and the single plastic-tip group. After brushing with dentifrice, the treated surfaces in all the treatment groups showed significantly fewer bacteria compared with the untreated surfaces in the control group, and the parts of the surfaces left untreated in the test groups. Within the limits of this study, treatment of titanium fixture surfaces with ultrasonic metal, plastic, or carbon tips significantly enhanced the bacterial removal efficacy of brushing. Thorough instrumentation that smooths the whole exposed surface may facilitate maintenance of the implants.

Modification of Structure and Properties of Titanium Surfaces During Formation of Silicides and Borides Initiated by High-Energy Treatment

An analysis of binary (Ti–Si, Ti–В, Si–B) and ternary (Ti–Si–B) phase states is made, their diagrams are presented, and a possibility for formation of a large number of metastable compounds is revealed. The latter are found to form as a result of application of non-equilibrium conditions in the course of material treatment with concentrated high-energy flows. Using an x-ray diffraction analysis and electron-diffraction microscopy, the phase composition of the surface layer of technical-grade titanium (VT1-0) treated by concentrated energy flows (irradiation with plasma from electrical wire explosion and high-intensity pulsed electron beam) is investigated.

Surface Analysis of Titanium Biological Modification with Glow Discharge

Glow discharge plasma (GDP) technology has been used to graft various proteins to the titanium surface, including albumin, type I collagen, but without fibronectin. The aim of this study was to evaluate and analyze the physical properties of fibronectin-grafted titanium surfaces after GDP treatment. Grade II titanium discs after cleaning and autoclaving were considered as original specimens, thus divided into four groups. The groups were different upon two treatments (GDP only and fibronectin grafting after GDP) and two storage temperature (4°C and 25°C). The implant surface morphology was characterized by scanning electron microscopy (SEM), roughness measurement, and wettability evaluation. The concentration relationship of fibronectin was by fluorescein isothiocyanate (FITC) labeling. SEM images showed that regular planar texture revealed on the surface of GDP-treated group, and irregular-folding protein was found on the fibronectin-grafted discs. Fibronectin-grafted groups had higher hydrophilicity and greater surface roughness than GDP-treated specimens. The storage temperature did not make obvious difference on the surface topography, wettability, and roughness. The number of fibronectin dots on the titanium surface labeling by FITC had positive relationship with the concentration of fibronectin solution used. Biologically modified titanium surface is more hydrophilic and rougher than GDP-treated ones. GDP treatment combined with fibronectin grafting increased the surface hydrophilicity and surface roughness of titanium discs, which may attribute to the affinity of cell adhesion, migration, proliferation, and differentiation.

Effects of Er: YAG laser irradiation on wettability, surface roughness, and biocompatibility of SLA titanium surfaces: an in vitro study

The erbium: yttrium-aluminum-garnet (Er: YAG) laser has been introduced as an effective method in the decontamination of implant surfaces. Data concerning the effects of the Er: YAG laser on the biological and surface properties of titanium are conflicting. Cellular behavior is greatly affected by surface properties, including composition, roughness, wettability, and morphology of the titanium surface. The aim of this study was to investigate the influence of the Er: YAG laser on the biocompatibility, surface roughness, and wettability of sandblasted and acid-etched (SLA) titanium surfaces. Twenty-one SLA titanium disks were irradiated by the Er: YAG laser at a pulse energy of 100mJ, with a pulse frequency of 10Hz under water irrigation for 1min. Cell viability, surface roughness, and wettability alterations were evaluated. Thirteen nonirradiated SLA disks were used as the control groups. Human osteoblast-like SaOs-2 cells were seeded onto the disks in culture media. Cell viability was evaluated using the methylthiazol tetrazolium assay. The surface roughness and wettability of the test and control groups were measured using profilometer and tensiometer devices, respectively. A significantly higher cell viability rate was observed in the test group (p = 0.032). The surface roughness was significantly reduced in the test group compared with the control group (p = 0.008). The surface wettability was significantly higher in the test group (p = 0.004). Within the limits of this study, the application of the Er: YAG laser with the previously described properties did not appear to have adverse effects on the biocompatibility of the SLA titanium surfaces. Application of this laser decreased the surface roughness and increased the wettability of the SLA titanium surfaces.

Hydroxyapatite synthesis on solid surfaces using a biological approach

Many naturally occurring mineralisation processes yield hydroxyapatite (HA) or related salts, but biological routes to calcification have not generally been exploited for production of hydroxyapatite for clinical and industrial applications. Serratia sp. NCIMB 40259 is a non-pathogenic Gram-negative bacterium which is capable of growing as a biofilm on many surfaces and can be used to form HA coatings on a variety of polymeric and metallic materials, including titanium. Here we review previous work and report the results of more recent studies on the influence of titanium compositional and surface properties on Serratia adherence and proliferation and biomineralisation on commercially pure titanium (cp Ti) discs and a Ti mesh. Bacterial adherence was equivalent on cpTi and Ti6Al4V, and biofilms formed on both rough and mirror-polished cpTi surfaces. Embedded alumina particles and alkali treatment did not noticeably alter the precipitation of Serratia HA, nor the structure of the coating in comparison with non-treated substrates. Coatings were retained after sintering at 800°C in argon, although the original curved plate-like crystals changed to nano-scale β-tricalcium phosphate particles. A phosphorous-rich diffusion zone formed at the coating-titanium interface. Bacterial mineralisation may have applications as a method for producing coatings on implants in non load-bearing sites, and non-clinical applications where a high surface area is the major concern.

In vitro Cyto and Blood Compatibility of Titanium Containing Diamond-Like Carbon Prepared by Hybrid Sputtering Method

In recent years, diamond-like carbon films (DLC) have been given more attention in research in the biomedical industry due to their potential application as surface coating on biomedical materials such as metals and polymer substrates. There are many ways to prepare metal containing DLC films deposited on polymeric film substrates, such as coatings from carbonaceous precursors and some means that incorporate other elements. In this study, we investigated both the surface and biocompatible properties of titanium containing DLC (Ti-DLC) films. The Ti-DLC films were prepared on the surface of poly (ethylene terephthalate) (PET) film as a function of the deposition power level using reactive sputtering technique. The films' hydrophilicity was studied by contact angle and surface energy tests. Their surface morphology was studied by scanning electron microscopy (SEM) and atomic force microscopy (AFM). Their elemental chemical composition was analyzed using energy dispersive X-spectra (EDX) and X-ray photoelectron spectroscopy (XPS). Their blood and cell compatibility was studied by in vitro tests, including tests on platelet adhesion, thrombus formation, whole blood clotting time and osteoblast cell compatibility. Significant changes in the morphological and chemical composition of the Ti-DLC films were observed and found to be a function of the deposition level. These morphological and chemical changes reduced the interfacial tension between Ti-DLC and blood proteins as well as resisted the adhesion and activation of platelets on the surface of the Ti-DLC films. The cell compatibility results exhibited significant growth of osteoblast cells on the surface of Ti incorporated DLC film compared with that of DLC film surface.

Surface characterization and biocompatibility of titanium alloys implanted with nitrogen by Hardion+ technology

In this study, the new Hardion+ micro-implanter technology was used to modify surface properties of biomedical pure titanium (CP-Ti) and Ti-6Al-4V ELI alloy by implantation of nitrogen ions. This process is based on the use of an electron cyclotron resonance ion source to produce a multienergetic ion beam from multicharged ions. After implantation, surface analysis methods revealed the formation of titanium nitride (TiN) on the substrate surfaces. An increase in superficial hardness and a significant reduction of friction coefficient were observed for both materials when compared to non-implanted samples. Better corrosion resistance and a significant decrease in ion release rates were observed for N-implanted biomaterials due to the formation of the protective TiN layer on their surfaces. In vitro tests performed on human fetal osteoblasts indicated that the cytocompatibility of N-implanted CP-Ti and Ti-6Al-4V alloy was enhanced in comparison to that of the corresponding non treated samples. Consequently, Hardion+ implantation technique can provide titanium alloys with better qualities in terms of corrosion resistance, cell proliferation, adhesion and viability.

Effects of storage methods on time-related changes of titanium surface properties and cellular response

Titanium implants are sold in the market as storable medical devices. All the implants have a certain shelf life during which they maintain their sterility, but variations of the surface properties through this duration have not been subject to a comprehensive assessment. The aim of this study was to investigate the effects of storage methods on time-related changes of titanium surface properties. Acid-etched titanium discs (Sa = 0.82 µm) were placed in a sealed container (tradition method) or submerged in the ddH2O/NaCl solution (0.15 mol L−1)/CaCl2 solution (0.15 mol L−1), and new titanium discs were used as a control group. SEM and optical profiler showed that surface morphology and roughness did not change within different groups, but the XPS analysis confirmed that the surface chemistry altered by different storage protocols as the storage duration increased, and the contact angle also varied with storage methods. The storage method also affected the protein adsorption capacity and cellular response on the titanium surface. All titanium discs stored in the solution maintained their excellent bioactivity even after four weeks storage time, but titanium discs stored in a traditional manner decreased substantially in an age-dependent manner. Much effort is needed to improve the storage methods in order to maintain the bioactivity of a titanium dental implant.