Satoshi Komasa

Osteogenic activity of titanium surfaces with nanonetwork structures

Background Titanium surfaces play an important role in affecting osseointegration of dental implants. Previous studies have shown that the titania nanotube promotes osseointegration by enhancing osteogenic differentiation. Only relatively recently have the effects of titanium surfaces with other nanostructures on osteogenic differentiation been investigated. Methods In this study, we used NaOH solutions with concentrations of 2.5, 5.0, 7.5, 10.0, and 12.5 M to develop a simple and useful titanium surface modification that introduces the nanonetwork structures with titania nanosheet (TNS) nanofeatures to the surface of titanium disks. The effects of such a modified nanonetwork structure, with different alkaline concentrations on the osteogenic differentiation of rat bone marrow mesenchymal stem cells (BMMSCs), were evaluated. Results The nanonetwork structures with TNS nanofeatures induced by alkali etching markedly enhanced BMMSC functions of cell adhesion and osteogenesis-related gene expression, and other cell behaviors such as proliferation, alkaline phosphatase activity, extracellular matrix deposition, and mineralization were also significantly increased. These effects were most pronounced when the concentration of NaOH was 10.0 M. Conclusion The results suggest that nanonetwork structures with TNS nanofeatures improved BMMSC proliferation and induced BMMSC osteogenic differentiation. In addition, the surfaces formed with 10.0 M NaOH suggest the potential to improve the clinical performance of dental implants.

Nanostructured Ti6Al4V alloy fabricated using modified alkali-heat treatment: Characterization and cell adhesion.

In order to optimize the creation of a nanostructured surface on Ti6Al4V titanium alloy, an alkali treatment was performed using a 10-M NaOH solution at various temperatures (30, 40, 50, and 60°C) so as to determine the optimal temperature. This was combined with subsequent heat treatments (200, 400, 600, and 800°C) in air. The effects of different temperatures for the latter treatments on the nanostructure surface and the initial cell adhesion were evaluated, and the optimal temperature of the alkali solution was found to be 30°C. Further, the nanotopography, surface chemistry, and surface roughness of the nanoporous structure were retained after heat treatments performed at 200, 400, and 600°C, and only the phase structure was altered. The amorphous sodium titanate phase, the content of which increased with increased heat-treatment temperature, may have played a role in promoting cell adhesion on the nanoporous surface. However, heat treatment at 800°C did not enhance the cell-surface attachment. Rather, the nanostructure degraded significantly with the reappearance of Al and V.

Synergistic effect of nanotopography and bioactive ions on peri-implant bone response

Both bioactive ion chemistry and nanoscale surface modifications are beneficial for enhanced osseointegration of endosseous implants. In this study, a facile synthesis approach to the incorporation of bioactive Ca2+ ions into the interlayers of nanoporous structures (Ca-nano) formed on a Ti6Al4V alloy surface was developed by sequential chemical and heat treatments. Samples with a machined surface and an Na+ ion-incorporated nanoporous surface (Na-nano) fabricated by concentrated alkali and heat treatment were used in parallel for comparison. The bone response was investigated by microcomputed tomography assessment, sequential fluorescent labeling analysis, and histological and histomorphometric evaluation after 8 weeks of implantation in rat femurs. No significant differences were found in the nanotopography, surface roughness, or crystalline properties of the Ca-nano and Na-nano surfaces. Bone–implant contact was better in the Ca-nano and Na-nano implants than in the machined implant. The Ca-nano implant was superior to the Na-nano implant in terms of enhancing the volume of new bone formation. The bone formation activity consistently increased for the Ca-nano implant but ceased for the Na-nano implant in the late healing stage. These results suggest that Ca-nano implants have promising potential for application in dentistry and orthopedics.

Effect of nanosheet surface structure of titanium alloys on cell differentiation

Titanium alloys are the most frequently used dental implants partly because of the protective oxide coating that spontaneously forms on their surface. We fabricated titania nanosheet (TNS) structures on titanium surfaces by NaOH treatment to improve bone differentiation on titanium alloy implants. The cellular response to TNSs on Ti6Al4V alloy was investigated, and the ability of the modified surfaces to affect osteogenic differentiation of rat bone marrow cells and increase the success rate of titanium implants was evaluated. The nanoscale network structures formed by alkali etching markedly enhanced the functions of cell adhesion and osteogenesis-related gene expression of rat bone marrow cells. Other cell behaviors, such as proliferation, alkaline phosphatase activity, osteocalcin deposition, and mineralization, were also markedly increased in TNS-modified Ti6Al4V. Our results suggest that titanium implants modified with nanostructures promote osteogenic differentiation, which may improve the biointegration of these implants into the alveolar bone.

Effect of ultraviolet treatment on bacterial attachment and osteogenic activity to alkali-treated titanium with nanonetwork structures

Purpose Alkali-treated titanium with nanonetwork structures (TNS) possesses good osteogenic activity; however, the resistance of this material to bacterial contamination remains inadequate. As such, TNS implants are prone to postoperative infection. In this work, we attempted to alter the biological properties of TNS by treatment with short-duration high-intensity ultraviolet (UV) irradiation. Methods TNS discs were treated with UV light (wavelength =254 nm, strength =100 mW/cm2) for 15 minutes using a UV-irradiation machine. We carried out a surface characterization and evaluated the discs for bacterial film formation, protein adsorption, and osteogenic features. Results The superhydrophilicity and surface hydrocarbon elimination exhibited by the treated material (UV-treated titanium with a nanonetwork structure [UV-TNS]) revealed that this treatment effectively changed the surface characteristics of TNS. Notably, UV-TNS also showed reduced colonization by Actinomyces oris during an initial attachment period and inhibition of biofilm formation for up to 6 hours. Moreover, compared to conventional TNS, UV-TNS showed superior osteogenic activity as indicated by increased levels of adhesion, proliferation, alkaline phosphatase activity, osteogenic factor production, and osteogenesis-related gene expression by rat bone marrow mesenchymal stem cells (rBMMSCs). This inverse relationship between bacterial attachment and cell adhesion could be due to the presence of electron–hole pairs induced by high-intensity UV treatment. Conclusion We suggest that simple UV treatment has great clinical potential for TNS implants, as it promotes the osseointegration of the TNS while reducing bacterial contamination, and can be conducted chair-side immediately prior to implantation.

Drug-Loadable Calcium Alginate Hydrogel System for Use in Oral Bone Tissue Repair.

This study developed a drug-loadable hydrogel system with high plasticity and favorable biological properties to enhance oral bone tissue regeneration. Hydrogels of different calcium alginate concentrations were prepared. Their swelling ratio, degradation time, and bovine serum albumin (BSA) release rate were measured. Human periodontal ligament cells (hPDLCs) and bone marrow stromal cells (BMSCs) were cultured with both calcium alginate hydrogels and polylactic acid (PLA), and then we examined the proliferation of cells. Inflammatory-related factor gene expressions of hPDLCs and osteogenesis-related gene expressions of BMSCs were observed. Materials were implanted into the subcutaneous tissue of rabbits to determine the biosecurity properties of the materials. The materials were also implanted in mandibular bone defects and then scanned using micro-CT. The calcium alginate hydrogels caused less inflammation than the PLA. The number of mineralized nodules and the expression of osteoblast-related genes were significantly higher in the hydrogel group compared with the control group. When the materials were implanted in subcutaneous tissue, materials showed favorable biocompatibility. The calcium alginate hydrogels had superior osteoinductive bone ability to the PLA. The drug-loadable calcium alginate hydrogel system is a potential bone defect reparation material for clinical dental application.

Bioactivity of NANOZR Induced by Alkali Treatment

In recent years, zirconia has been a recognized implant material in clinical dentistry. In the present study, we investigated the performance of an alkali-modified ceria-stabilized tetragonal ZrO2 polycrystalline ceramic-based nanostructured zirconia/alumina composite (NANOZR) implant by assessing surface morphology and composition, wettability, bovine serum albumin adsorption rate, rat bone marrow (RBM) cell attachment, and capacity for inducing bone differentiation. NANOZR surfaces without and with alkali treatment served as the control and test groups, respectively. RBM cells were seeded in a microplate with the implant and cultured in osteogenic differentiation medium, and their differentiation was evaluated by measuring alkaline phosphatase (ALP) activity, osteocalcin (OCN) production, calcium deposition, and osteogenic gene expression. The alkali-treated NANOZR surface increased ALP activity, OCN production, calcium deposition, and osteogenesis-related gene expression in attached RBM cells. These data suggest that alkali treatment enhances the osteogenesis-inducing capacity of NANOZR implants and may therefore improve their biointegration into alveolar bone.

Analysis of Titania Nanosheet Adsorption Behavior Using a Quartz Crystal Microbalance Sensor

We investigated the adsorption of albumin and fibronectin on a titania nanosheet- (TNS-) modified quartz crystal microbalance (QCM) sensor. A Ti QCM sensor was fabricated by reactive magnetron sputtering. A thin layer of Ti was deposited on the QCM sensor. This sensor was then alkali-modified by treatment with NaOH at room temperature to fabricate the titania nanosheets. Scanning probe microscopy, X-ray photoelectron spectroscopy, and scanning electron microscopy were performed to investigate the surface topology and chemical components of each sensor. The TNS had a titanium oxide film exhibiting a nodular structure and a thickness of 13 nm on the QCM sensor. Furthermore, QCM measurements showed significantly greater amounts of albumin and fibronectin adsorbed on the TNS than on titanium. The NaOH treatment of titanium modified the sensor surface and improved the adsorption behaviors of proteins related to the initial adhesion of bone marrow cells. Therefore, we concluded that TNS improves the initial adhesion between the implant materials and the surrounding tissues.

Effects of glucose concentration on osteogenic differentiation of type II diabetes mellitus rat bone marrow-derived mesenchymal stromal cells on a nano-scale modified titanium

BACKGROUND AND OBJECTIVE Diabetes mellitus (DM) is a common disease worldwide. Patients with DM have an increased risk of losing their teeth compared with other individuals. Dental implants are a standard of care for treating partial or full edentulism, and various implant surface treatments have recently been developed to increase dental implant stability. However, some studies have reported that DM reduces osseointegration and the success rate of dental implants. The purpose of this study was to determine the effects of high glucose levels for hard tissue formation on a nano-scale modified titanium surface. MATERIAL AND METHODS Titanium disks were heated at 600°C for 1 h after treatment with or without 10 m NaOH solution. All disks were incubated with type II DM rat bone marrow-derived mesenchymal stromal cells before exposure to one of four concentrations of glucose (5.5, 8.0, 12.0 or 24.0 mm). The effect of different glucose concentrations on bone marrow-derived mesenchymal stromal cell osteogenesis and inflammatory cytokines on the nano-scale modified titanium surface was evaluated. RESULTS Alkaline phosphatase activity decreased with increasing glucose concentration. In contrast, osteocalcin production and calcium deposition were significantly decreased at 8.0 mm glucose, but increased with glucose concentrations over 8.0 mm. Differences in calcium/phosphate ratio associated with the various glucose concentrations were similar to osteocalcin production and calcium deposition. Inflammatory cytokines were expressed at high glucose concentrations, but the nano-scale modified titanium surface inhibited the effect of high glucose concentrations. CONCLUSION High glucose concentration increased hard tissue formation, but the quality of the mineralized tissue decreased. Furthermore, the nano-scale modified titanium surface increased mineralized tissue formation and anti-inflammation, but the quality of hard tissue was dependent on glucose concentration.

Biocompatibility of titanium surface nanostructures following chemical processing and heat treatment

The aims of the present study were to investigate alkali and heat treatment of titanium surfaces, and to evaluate the ability of such modified surfaces to improve osteogenic differentiation of rat bone marrow (RBM) cells to increase the success rate of titanium implants. To improve bone differentiation on titanium alloy implants, titania nanosheet (TNS) structures were fabricated on titanium surfaces by NaOH and heat treatment at 200, 400, 600, and 800°C. Cell behavior on heattreated TNS-modified titanium was investigated, and the ability of the modified surfaces to affect osteogenic differentiation of RBM cells and to increase the success rate of titanium implants was evaluated. The nanoscale network structures formed by alkali etching markedly enhanced the cell adhesion and osteogenesis-related gene expression of RBM cells. Other cell behaviors, such as proliferation, alkaline phosphatase activity, osteocalcin deposition, and mineralization, were also markedly increased on heat-treated TNS-modified titanium. Our results suggest that nanostructure-modified titanium implants promote osteogenic differentiation, which may improve biointegration of these implants into alveolar bone.