Sand-blasted And Acid-etched Titanium Research Articles

Piranha-etched titanium nanostructure reduces biofilm formation in vitro

ObjectivesNano-modified surfaces for dental implants may improve gingival fibroblast adhesion and antibacterial characteristics through cell-surface interactions. The present study investigated how a nanocavity titanium surface impacts the viability and adhesion of human gingival fibroblasts (HGF-1) and compared its response to Porphyromonas gingivalis with those of marketed implant surfaces.Material and methodsCommercial titanium and zirconia disks, namely, sandblasted and acid-etched titanium (SLA), sandblasted and acid-etched zirconia (ZLA), polished titanium (PT) and polished zirconia (ZrP), and nanostructured disks (NTDs) were tested. Polished titanium disks were etched with a 1:1 combination of 98% H2SO4 and 30% H2O2 (piranha etching) for 5 h at room temperature to produce the NTDs. Atomic force microscopy was used to measure the surface topography, roughness, adhesion force, and work of adhesion. MTT assays and immunofluorescence staining were used to examine cell viability and adhesion after incubation of HGF-1 cells on the disk surfaces. After incubation with P. gingivalis, conventional culture, live/dead staining, and SEM were used to determine the antibacterial properties of NTD, SLA, ZLA, PT, and ZrP.ResultsEtching created nanocavities with 10–20-nm edge-to-edge diameters. Chemical etching increased the average surface roughness and decreased the surface adherence, while polishing and flattening of ZrP increased adhesion. However, only the NTDs inhibited biofilm formation and bacterial adherence. The NTDs showed antibacterial effects and P. gingivalis vitality reductions. The HGF-1 cells demonstrated greater viability on the NTDs compared to the controls.ConclusionNanocavities with 10–20-nm edge-to-edge diameters on titanium disks hindered P. gingivalis adhesion and supported the adhesion of gingival fibroblasts when compared to the surfaces of currently marketed titanium or zirconia dental implants.Clinical relevanceThis study prepared an effective antibacterial nanoporous surface, assessed its effects against oral pathogens, and demonstrated that surface characteristics on a nanoscale level influenced oral pathogens and gingival fibroblasts.Clinical trial registration: not applicable

Effect of photodynamic therapy according to differences in photosensitizers on Staphylococcus aureus biofilm on titanium.

This study aimed to evaluate the antimicrobial effect of photodynamic therapy (PDT) against Staphylococcus aureus biofilm on a titanium surface and to compare the differences in the effect of PDT using toluidine blue O (TBO) and methylene blue (MB) as a photosensitizer. The bacterial strain S. aureus ATCC 25,923 was used. Sandblasted and acid-etched (SLA) disks were divided into the following six groups: phosphate buffer saline (PBS), TBO, MB, PBS with laser (PBS+L), TBO with laser (TBO+L), and MB with laser (MB+L). The laser group samples were irradiated by a cold diode laser for 60s. After treatment, the number of surviving bacteria was calculated by counting the colony-forming units (CFUs) and confocal laser scanning microscopy (CLSM) was applied to observe the bacteria on the disk surface. The TBO+L and MB+L groups showed significantly lower CFU/ml than the other groups (p < 0.01). The TBO+L group showed significantly lower CFU/ml than the MB+L group (p=0.032). There was no significant difference between the PBS, TBO, MB, and PBS+L groups. Within the limitations of this in vitro study, PDT with TBO and MB can effectively reduce S. aureus biofilm on SLA titanium surfaces. TBO is more effective than MB as a photosensitizer. PDT with TBO may be applied to the treatment of peri‑implant disease in the future.

Alterations in Surface Roughness and Chemical Characteristics of Sandblasted and Acid-Etched Titanium Implants after Irradiation with Different Diode Lasers

The purpose of this study was to evaluate the effects of diode laser irradiation with different wavelengths on the surface roughness (Ra) and chemical composition of sandblasted and acid-etched (SLA) titanium implants. Three types of diode lasers with different wavelengths were irradiated on the titanium implants at output powers of 1.0, 2.0, and 3.0 W. The mean Ra values for all spots were measured using a scanning probe microscope. Analysis of variance tests were performed to verify the differences in the Ra between groups according to the type of lasers or power out (α = 0.05). For analyzing chemical composition, atomic and weight percent ratios of titanium, oxygen, and carbon were measured using energy-dispersive spectrometry (EDS). The mean Ra of titanium disc was higher in the 3.0-W output than in 1.0-W or 2.0-W output, but there was no statistically significant difference (p &gt; 0.05). In EDS analysis, it was difficult to find a clear difference in the titanium, oxygen, and carbon element ratios between the laser-irradiated and nonirradiated groups. The irradiation of diode laser with 1.0, 2.0, and 3.0 W output for 15 s decontaminated the SLA titanium surface without damage. However, additional clinical trials will be needed to verify the results of the present study.

The bactericidal effect of an atmospheric-pressure plasma jet on Porphyromonas gingivalis biofilms on sandblasted and acid-etched titanium discs.

PurposeDirect application of atmospheric-pressure plasma jets (APPJs) has been established as an effective method of microbial decontamination. This study aimed to investigate the bactericidal effect of direct application of an APPJ using helium gas (He-APPJ) on Porphyromonas gingivalis biofilms on sandblasted and acid-etched (SLA) titanium discs.MethodsOn the SLA discs covered by P. gingivalis biofilms, an APPJ with helium (He) as a discharge gas was applied at 3 different time intervals (0, 3, and 5 minutes). To evaluate the effect of the plasma itself, the He gas–only group was used as the control group. The bactericidal effect of the He-APPJ was determined by the number of colony-forming units. Bacterial viability was observed by confocal laser scanning microscopy (CLSM), and bacterial morphology was examined by scanning electron microscopy (SEM).ResultsAs the plasma treatment time increased, the amount of P. gingivalis decreased, and the difference was statistically significant. In the SEM images, compared to the control group, the bacterial biofilm structure on SLA discs treated by the He-APPJ for more than 3 minutes was destroyed. In addition, the CLSM images showed consistent results. Even in sites distant from the area of direct He-APPJ exposure, decontamination effects were observed in both SEM and CLSM images.ConclusionsHe-APPJ application was effective in removing P. gingivalis biofilm on SLA titanium discs in an in vitro experiment.

The Effects of a Er:YAG Laser on Machined, Sand-Blasted and Acid-Etched, and Resorbable Blast Media Titanium Surfaces Using Confocal Microscopy and Scanning Electron Microscopy

Purpose: Laser treatment has become a popular method in implant dentistry, and lasers have been used for the decontamination of implant surfaces when treating peri-implantitis. This study was performed to evaluate the effects of an Erbium-doped:Yttrium-Aluminum-Garnet (Er:YAG) laser with different settings on machined (MA), sand-blasted and acid-etched (SA), and resorbable blast media (RBM) titanium surfaces using scanning electron microscopy and confocal microscopy. Materials and Methods: Four MA, four SA, and four RBM discs were either irradiated at 40 mJ/20 Hz, 90 mJ/20 Hz, or 40 mJ/25 Hz for 2 minutes. The specimens were evaluated with scanning electron microscopy and confocal microscopy. Result: The untreated MA surface demonstrated uniform roughness with circumferential machining marks, and depressions were observed after laser treatment. The untreated SA surface demonstrated a rough surface with sharp spikes and deep pits, and the laser produced noticeable changes on the SA titanium surfaces with melting and fusion. The untreated RBM surface demonstrated a rough surface with irregular indentation, and treatment with the laser produced changes on the RBM titanium surfaces. The Er:YAG laser produced significant changes on the roughness parameters, including arithmetic mean height of the surface (Sa) and maximum height of the surface (Sz), of the MA and SA surfaces. However, the Er:YAG laser did not produce notable changes on the roughness parameters, such as Sa and Sz, of the RBM surfaces. Conclusion: This study evaluated the effects of an Er:YAG laser on MA, SA, and RBM titanium discs using confocal microscopy and scanning electron microscopy. Treatment with the laser produced significant changes in the roughness of MA and SA surfaces, but the roughness parameters of the RBM discs were not significantly changed. Further research is needed to evaluate the efficiency of the Er:YAG laser in removing the contaminants, adhering bacteria, and the effects of treatment on cellular attachment, proliferation, and differentiation.

The Effect of Platelet Proteins Released in Response to Titanium Implant Surfaces on Macrophage Pro-Inflammatory Cytokine Gene Expression.

Platelets are one of the earliest cell types to interact with surgically inserted titanium implants. This in vitro study investigated the effect of titanium surface-induced platelet releasate on macrophage cytokine gene expression. To mimic the in vivo temporal sequence of platelet arrival and protein production at the implant surface and the subsequent effect of these proteins on mediators of the immune response, the levels of platelet attachment and activation in response to culture on smooth polished, sandblasted and acid-etched (SLA), and hydrophilic-modified SLA (modSLA) titanium surfaces were first determined by microscopy and protein assay. The subsequent effect of the platelet-released proteins on human THP-1 macrophage cytokine gene expression was determined by polymerase chain reaction array after 1 and 3 days of macrophage culture on the titanium surfaces in platelet-releasate conditioned media. Platelet attachment was surface dependent with decreased attachment observed on the hydrophilic (modSLA) surface. The platelet releasate, when considered independently of the surface effect, elicited an overall pro-inflammatory response in macrophage cytokine gene expression, that is, the expression of typical pro-inflammatory cytokine genes such as TNF, IL1a, IL1b, and CCL1 was significantly up-regulated whereas the expression of anti-inflammatory cytokine genes such as IL10, CxCL12, and CxCL13 was significantly down-regulated. However, following platelet exposure to different surface modifications, the platelet releasate significantly attenuated the macrophage pro-inflammatory response to microrough (SLA) titanium and hastened an anti-inflammatory response to hydrophilic (modSLA) titanium. Theses results demonstrate that titanium surface topography and chemistry are able to influence the proteomic profile released by platelets, which can subsequently influence macrophage pro-inflammatory cytokine expression. This immunomodulation may be an important mechanism via which titanium surface modification influences osseointegration.

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.

Treatment With Various Ultrasonic Scaler Tips Affects Efficiency of Brushing of SLA Titanium Discs

The dental implant surface will be colonized by bacteria once it is exposed to the oral cavity. It is necessary to keep the titanium surface clean to prevent peri-implant diseases. Mechanical instrumentation is widely used, but this may cause damage to the implant surfaces. There is limited information whether surface change resulting from instrumentation influences the adherence of bacteria to the implant surface or influences the ease of removal of bacteria from the titanium surface by daily brushing. Therefore, this in vitro study was performed (1) to evaluate removal of Porphyromonas gingivalis from sand-blasted and acid-etched (SLA) titanium discs after the discs were instrumented by various ultrasonic scaler tips or brushed with a toothbrush with dentifrice using crystal violet assay and scanning electron microscopy (SEM), and (2) to assess the change of surface roughness after the treated discs were brushed with a toothbrush with dentifrice. SLA discs were treated with various ultrasonic scaler tips and a toothbrush. The titanium discs were incubated with P. gingivalis for 2 days after treatment (ultrasonic scales tips and brush) and then the disc surfaces were brushed for total of 40 seconds (20 seconds, two cycles) with a toothbrush with dentifrice. Differences in adhering bacteria were evaluated using crystal violet assay and SEM. Surface roughness of the treated discs after brushing with dentifrice was measured using confocal microscopy. The change of surface structure was observed after different treatment modalities. Removal of bacteria was increased with the longer time of brushing, and the ultrasonic metal tip group displayed a significantly lower number of bacteria after brushing when compared to other groups. Within the limits of this study, it may be suggested that when SLA surface is exposed to the oral cavity, it should firstly be treated with metal tips to smoothen the rough surface and thereby reduce attachment of bacteria and facilitate the removal of bacteria by daily oral hygiene procedures.

An in vitro evaluation of the responses of human osteoblast-like SaOs-2 cells to SLA titanium surfaces irradiated by erbium:yttrium–aluminum–garnet (Er:YAG) lasers

Erbium:yttrium-aluminum-garnet (Er:YAG) laser treatment is an effective option for the removal of bacterial plaques. Many studies have shown that Er:YAG lasers cannot re-establish the biocompatibility of titanium surfaces. The aim of this study was to evaluate the responses of the human osteoblast-like cell line, SaOs-2, to sand-blasted and acid-etched (SLA) titanium surface irradiation using different energy settings of an Er:YAG laser by examining cell viability and morphology. Forty SLA titanium disks were irradiated with an Er:YAG laser at a pulse energy of either 60 or 100 mJ with a pulse frequency of 10 Hz under water irrigation and placed in a 24-well plate. Human osteoblast-like SaOs-2 cells were seeded onto the disks in culture media. Cells were then kept in an incubator with 5% carbon dioxide at 37 °C. Each experimental group was divided into two smaller groups to evaluate cell morphology by scanning electron microscope and cell viability using 3-4,5-dimethylthiazol 2,5-diphenyltetrazolium bromide test. In both the 60 and the 100 mJ experimental groups, spreading morphologies, with numerous cytoplasmic extensions, were observed prominently. Similarly, a majority of cells in the control group exhibited spreading morphologies with abundant cytoplasmic extensions. There were no significant differences among the laser and control groups. The highest cell viability rate was observed in the 100 mJ laser group. No significant differences were observed between the cell viability rates of the two experimental groups (p = 1.00). In contrast, the control group was characterized by a significantly lower cell viability rate (p < 0.001). Treatments with an Er:YAG laser at a pulse energy of either 60 or 100 mJ do not reduce the biocompatibility of SLA titanium surfaces. In fact, modifying SLA surfaces with Er:YAG lasers improved the biocompatibility of these surfaces.

Evaluation of the attachment, proliferation, and differentiation of osteoblast on a calcium carbonate coating on titanium surface

Titanium has been reported to have some limitations in dental and orthopaedic clinical application. This study described a coating process using a simple chemical method to prepare calcium carbonate coatings on smooth titanium (STi) and sandblasted and acid-etched titanium (SATi), and evaluated the biological response of the materials in vitro. The surfaces of STi, SATi, calcium carbonate coated STi (CC-STi) and calcium carbonate coated SATi (CC-SATi) were characterized for surface roughness, contact angles, surface morphology and surface chemistry. The morphology of MG63 cells cultured on the surfaces was observed by SEM and Immuno-fluorescence staining. Cell attachment/proliferation was assessed by MTT assay, and cell differentiation was evaluated by alkaline phosphatase (ALP) activity. MG63 was found to attach favorably to calcium carbonate crystals with longer cytoplasmic extensions on CC-STi and CC-SATi, resulting in lower cell proliferation but higher ALP activity when compared to STi and SATi respectively. Moreover, CC-SATi is more favorable than CC-STi in terms of biological response. In conclusion, the calcium carbonate coatings on titanium were supposed to improve the osteointegration process and stimulate osteoblast differentiation, especially in early stage. And this method could possibly be a feasible alternative option for future clinical application.

One-step modification of nano-hydroxyapatite coating on titanium surface by hydrothermal method

Since metallic biomaterials used for orthopedic and dental implants possess a paucity of reactive functional groups, bioactivity modification of these materials is challenging. To enhance osteoconductivity, sandblasted and acid etched titanium (SLA-Ti) discs were hydrothermally treated in simple suspension of hydroxyapatite (HA) by a one-step method in the present work. Change of surface chemistry, surface morphology of samples, and structure of formed nano-HA were investigated by X-ray photoelectron spectrometry (XPS), scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR) and X-ray powder diffraction (XRD). The results demonstrated that the concentration of hydroxyl groups increased after hydrothermal treatment. The ability of the titanium surface to form nano-HA was enhanced by increasing the hydrothermal treatment temperature and time. The bonding strength of the formed nano-HA coating, examined by an ultrasonic cleaning process, was higher than that of HA coating generated directly by HA suspension deposition. Culturing of MC3T3-E1 cells on the SLA-Ti and HT-Ti discs in vitro showed that HT-Ti was more favorable for cell attachment and differentiation than SLA-Ti. Hence the one-step hydrothermal treatment with HA suspension was a simple, effective and promising method for HA coating modification of titanium implants.

Literature Reviews

Literature Reviews

Effect of enamel matrix protein derivative on the attachment, proliferation, and viability of human SaOs(2) osteoblasts on titanium implants.

The purpose of the present study was to investigate the effects of an enamel matrix protein derivative (EMD) on attachment, proliferation, and viability of human SaOs(2) osteoblasts on titanium implants. A total of 220 sand-blasted and acid-etched (SLA) titanium discs were placed into 24-well culture plates. Before cell inoculation, McCoy's 5A medium (MCM) containing EMD at 25 micro g/ml, 50 micro g/ml, 100 micro g/ml, and 200 micro g/ml was added, and the culture plates were incubated for 30 min. As control, MCM alone was used. Human osteoblast-like cells (SaOs(2)) (2 x 10(4) cells, fourth passage) were suspended in MCM containing 1% penicillin/streptomycin and 10% fetal bovine serum and then inoculated into the well chambers. The medium was changed after 3 days without the addition of EMD. At days 1, 3, and 6, DNA content of the cells was assessed using the CyQuant cell proliferation assay kit, and mitochondrial activity of the cells was measured using a CellTiter-Glo luminescent cell viability assay. The presence of EMD on the titanium discs at days 1 and 6 was evaluated using immunofluorescence stain (IFS) by means of polyclonal antibodies against amelogenin. Additionally, cell morphology was investigated using scanning electron microscopy. Enamel matrix derivative at 25 micro g/ml, 50 micro g/ml, 100 micro g/ml, and 200 micro g/ml demonstrated similar increases in cell proliferation as the control medium at days 3 and 6 ( P>0.05 between groups, respectively). Proliferation, however, appeared to be ameliorated with increasing EMD concentrations. At 25 micro g/ml and 50 micro g/ml, EMD also demonstrated an increase in cell viability similar to the control medium at days 3 and 6 ( P>0.05 between groups, respectively), while EMD at 100 micro g/ml and 200 micro g/ml resulted in statistically significant higher increase in cell viability than in the control medium at day 6 ( P<0.001 between groups, respectively). In all test groups, IFS at day 6 was markedly lower than at day 1. Scanning electron microscopy revealed comparable cell morphology in all groups. Within the limits of the present study, it was concluded that EMD enhanced cell proliferation and viability of human SaOs(2) osteoblasts on SLA titanium implants in a concentration-dependent manner.

Bone response to unloaded and loaded titanium implants with a sandblasted and acid-etched surface: a histometric study in the canine mandible.

Many dental clinical implant studies have focused on the success of endosseous implants with a variety of surface characteristics. Most of the surface alterations have been aimed at achieving greater bone-to-implant contact as determined histometrically at the light microscopic level. A previous investigation in non-oral bone under short-term healing periods (3 and 6 weeks) indicated that a sandblasted and acid-etched titanium (SLA) implant had a greater bone-to-implant contact than did a comparably-shaped implant with a titanium plasma-sprayed (TPS) surface. In this canine mandible study, nonsubmerged implants with a SLA surface were compared to TPS-coated implants under loaded and nonloaded conditions for up to 15 months. Six foxhound dogs had 69 implants placed in an alternating pattern with six implants placed bilaterally in each dog. Gold crowns that mimicked the natural occlusion were fabricated for four dogs. Histometric analysis of bone contact with the implants was made for two dogs after 3 months of healing (unloaded group), 6 months of healing (3 months loaded), and after 15 months of healing (12 months loaded). The SLA implants had a significantly higher (p < 0.001) percentage of bone-to-implant contact than did the TPS implants after 3 months of healing (72.33 +/- 7.16 versus 52.15 +/- 9.19; mean +/- SD). After 3 months of loading (6 months of healing) no significant difference was found between the SLA and TPS surfaced implants (68.21 +/- 10.44 and 78.18 +/- 6.81, respectively). After 12 months of loading (15 months of healing) the SLA implants had a significantly greater percentage (p < 0.001) of bone-to-implant contact than did the TPS implants (71.68 +/- 6.64 and 58.88 +/- 4.62, respectively). No qualitative differences in bone tissue were observed between the two groups of implants nor was there any difference between the implants at the clinical level. These results are consistent with earlier studies on SLA implants and suggest that this surface promotes greater osseous contact at earlier time points compared to TPS-coated implants.